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Sample records for campylobacter phages including

  1. Characterization of Campylobacter phages including analysis of host range by selected Campylobacter Penner serotypes

    DEFF Research Database (Denmark)

    Hansen, Vinni; Rosenquist, Hanne; Baggesen, Dorte Lau

    2007-01-01

    Background: The predominant food borne pathogen in the western world today is Campylobacter. Campylobacter specific bacteriophages (phages) have been proposed as an alternative agent for reducing the burden of Campylobacter in broilers. One concern in relation to phage biocontrol is the narrow host...... range often displayed by phages. To identify the potential of phages as a Campylobacter reducing agent we needed to determine their infectivity on a panel of isolates representing the Campylobacter strains found in broilers as well as humans. Results: In this study, Campylobacter phages were isolated...... from the intestines of broilers and ducks and from abattoir sewage. Twelve phages were investigated to determine their ability to infect the Campylobacter Penner serotypes commonly present in Danish poultry and patients with campylobacteriosis. A total of 89% of the Campylobacter jejuni strains and 14...

  2. Use of phages to control Campylobacter spp.

    Science.gov (United States)

    Janež, Nika; Loc-Carrillo, Catherine

    2013-10-01

    The use of phages to control pathogenic bacteria has been investigated since they were first discovered in the beginning of the 1900s. Over the last century we have slowly gained an in-depth understanding of phage biology including which phage properties are desirable when considering phage as biocontrol agents and which phage characteristics to potentially avoid. Campylobacter infections are amongst the most frequently encountered foodborne bacterial infections around the world. Handling and consumption of raw or undercooked poultry products have been determined to be the main route of transmission. The ability to use phages to target these bacteria has been studied for more than a decade and although we have made progress towards deciphering how best to use phages to control Campylobacter associated with poultry production, there is still much work to be done. This review outlines methods to improve the isolation of these elusive phages, as well as methods to identify desirable characteristics needed for a successful outcome. It also highlights the body of research undertaken so far and what criteria to consider when doing in-vivo studies, especially because some in-vitro studies have not been found to translate into to phage efficacy in-vivo.

  3. Phage therapy reduces Campylobacter jejuni colonization in broilers

    NARCIS (Netherlands)

    Wagenaar, J.A.; Bergen, van M.A.P.; Mueller, M.A.; Wassenaar, T.M.; Carlton, R.M.

    2005-01-01

    The effect of phage therapy in the control of Campylobacter jejuni colonization in young broilers, either as a preventive or a therapeutic measure, was tested. A prevention group was infected with C. jejuni at day 4 of a 10-day phage treatment. A therapeutic group was phage treated for 6 days, start

  4. Phage therapy reduces Campylobacter jejuni colonization in broilers

    NARCIS (Netherlands)

    Wagenaar, J.A.; Bergen, van M.A.P.; Mueller, M.A.; Wassenaar, T.M.; Carlton, R.M.

    2005-01-01

    The effect of phage therapy in the control of Campylobacter jejuni colonization in young broilers, either as a preventive or a therapeutic measure, was tested. A prevention group was infected with C. jejuni at day 4 of a 10-day phage treatment. A therapeutic group was phage treated for 6 days,

  5. Colonisation of a phage susceptible Campylobacter jejuni population in two phage positive broiler flocks.

    Directory of Open Access Journals (Sweden)

    Sophie Kittler

    Full Text Available The pathogens Campylobacter jejuni and Campylobacter coli are commensals in the poultry intestine and campylobacteriosis is one of the most frequent foodborne diseases in developed and developing countries. Phages were identified to be effective in reducing intestinal Campylobacter load and this was evaluated, in the first field trials which were recently carried out. The aim of this study was to further investigate Campylobacter population dynamics during phage application on a commercial broiler farm. This study determines the superiority in colonisation of a Campylobacter type found in a field trial that was susceptible to phages in in vitro tests. The colonisation factors, i.e. motility and gamma glutamyl transferase activity, were increased in this type. The clustering in phylogenetic comparisons of MALDI-TOF spectra did not match the ST, biochemical phenotype and phage susceptibility. Occurrence of Campylobacter jejuni strains and phage susceptibility types with different colonisation potential seem to play a very important role in the success of phage therapy in commercial broiler houses. Thus, mechanisms of both, phage susceptibility and Campylobacter colonisation should be further investigated and considered when composing phage cocktails.

  6. Primary Isolation Strain Determines Both Phage Type and Receptors Recognised by Campylobacter jejuni Bacteriophages

    DEFF Research Database (Denmark)

    Sørensen, Martine C. Holst; Gencay, Yilmaz Emre; Birk, Tina;

    2015-01-01

    In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated...

  7. Significance of phage-host interactions for biocontrol of Campylobacter jejuni in food

    DEFF Research Database (Denmark)

    Athina, Zampara; Sørensen, Martine Camilla Holst; Elsser-Gravesen, Anne

    2017-01-01

    Poultry meat is the main source of Campylobacter jejuni foodborne disease. Currently, no effective control measures prevent C. jejuni from contaminating poultry meat. However, post-harvest phage treatment is a promising biocontrol strategy that has not yet been explored. Here we identified phages...... capable of reducing C. jejuni at chilled temperature by a systematic screening of unique phages of our collection consisting of flagellotropic phages and phages dependent on capsular polysaccharides (CPSs) for infection. Interestingly, CPS phages showed varied killing efficiencies at 5 °C in vitro......, ranging from insignificant reduction to 0.55 log reduction. In contrast, none of the flagellotropic phages significantly reduced C. jejuni counts at low temperature. Phage adsorption at 5 °C showed that flagellotropic phages bind reversibly and less efficiently to C. jejuni than CPS phages, which may...

  8. Primary Isolation Strain Determines Both Phage Type and Receptors Recognised by Campylobacter jejuni Bacteriophages

    DEFF Research Database (Denmark)

    Sørensen, Martine C. Holst; Gencay, Yilmaz Emre; Birk, Tina

    2015-01-01

    In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated...... using NCTC12662 as the indicator strain, which may have biased the selection of phages. A large group of C. jejuni phages rely on the highly diverse capsular polysaccharide (CPS) for infection and recent work identified the O-methyl phosphoramidate modification (MeOPN) of CPS as a phage receptor. We...... therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages...

  9. Primary Isolation Strain Determines Both Phage Type and Receptors Recognised by Campylobacter jejuni Bacteriophages

    DEFF Research Database (Denmark)

    Sørensen, Martine C. Holst; Gencay, Yilmaz Emre; Birk, Tina;

    2015-01-01

    were identified based on host range analysis and genome restriction profiles. Most phages were isolated using C. jejuni strains NCTC12662 and RM1221 and interestingly phage genome size (140 kb vs. 190 kb), host range and morphological appearance correlated with the isolation strain. Thus, according......In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated...... therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages...

  10. Primary isolation strain determines both phage type and receptors recognised by Campylobacter jejuni bacteriophages.

    Science.gov (United States)

    Sørensen, Martine C Holst; Gencay, Yilmaz Emre; Birk, Tina; Baldvinsson, Signe Berg; Jäckel, Claudia; Hammerl, Jens A; Vegge, Christina S; Neve, Horst; Brøndsted, Lone

    2015-01-01

    In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated using NCTC12662 as the indicator strain, which may have biased the selection of phages. A large group of C. jejuni phages rely on the highly diverse capsular polysaccharide (CPS) for infection and recent work identified the O-methyl phosphoramidate modification (MeOPN) of CPS as a phage receptor. We therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages were identified based on host range analysis and genome restriction profiles. Most phages were isolated using C. jejuni strains NCTC12662 and RM1221 and interestingly phage genome size (140 kb vs. 190 kb), host range and morphological appearance correlated with the isolation strain. Thus, according to C. jejuni phage grouping, NCTC12662 and NCTC12658 selected for CP81-type phages, while RM1221 selected for CP220-type phages. Furthermore, using acapsular ∆kpsM mutants we demonstrated that phages isolated on NCTC12658 and NCTC12662 were dependent on the capsule for infection. In contrast, CP220-type phages isolated on RM1221 were unable to infect non-motile ∆motA mutants, hence requiring motility for successful infection. Hence, the primary phage isolation strain determines both phage type (CP81 or CP220) as well as receptors (CPS or flagella) recognised by the isolated phages.

  11. Primary isolation strain determines both phage type and receptors recognised by Campylobacter jejuni bacteriophages.

    Directory of Open Access Journals (Sweden)

    Martine C Holst Sørensen

    Full Text Available In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated using NCTC12662 as the indicator strain, which may have biased the selection of phages. A large group of C. jejuni phages rely on the highly diverse capsular polysaccharide (CPS for infection and recent work identified the O-methyl phosphoramidate modification (MeOPN of CPS as a phage receptor. We therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages were identified based on host range analysis and genome restriction profiles. Most phages were isolated using C. jejuni strains NCTC12662 and RM1221 and interestingly phage genome size (140 kb vs. 190 kb, host range and morphological appearance correlated with the isolation strain. Thus, according to C. jejuni phage grouping, NCTC12662 and NCTC12658 selected for CP81-type phages, while RM1221 selected for CP220-type phages. Furthermore, using acapsular ∆kpsM mutants we demonstrated that phages isolated on NCTC12658 and NCTC12662 were dependent on the capsule for infection. In contrast, CP220-type phages isolated on RM1221 were unable to infect non-motile ∆motA mutants, hence requiring motility for successful infection. Hence, the primary phage isolation strain determines both phage type (CP81 or CP220 as well as receptors (CPS or flagella recognised by the isolated phages.

  12. Impact of a Single Phage and a Phage Cocktail Application in Broilers on Reduction of Campylobacter jejuni and Development of Resistance

    Science.gov (United States)

    Fischer, Samuel; Kittler, Sophie; Klein, Günter; Glünder, Gerhard

    2013-01-01

    Campylobacteriosis is currently the most frequent foodborne zoonosis in many countries. One main source is poultry. The aim of this study was to enhance the knowledge about the potential of bacteriophages in reducing colonization of broilers with Campylobacter , as there are only a few in vivo studies published. Commercial broilers were inoculated with 104 CFU/bird of a Campylobacter jejuni field strain. Groups of 88 birds each were subsequently treated with a single phage or a four-phage cocktail (107 PFU/bird in CaCO3 buffered SM-Buffer). Control birds received the solvent only. Afterwards, subgroups of eleven birds each were examined for their loads with phages and Campylobacter on day 1, 3, 7, 14, 21, 28, 35 and 42 after phage application. The susceptibility of the Campylobacter population to phage infection was determined using ten isolates per bird. In total 4180 re-isolates were examined. The study demonstrated that the deployed phages persisted over the whole investigation period. The Campylobacter load was permanently reduced by the phage-cocktail as well as by the single phage. The reduction was significant between one and four weeks after treatment and reached a maximum of log10 2.8 CFU/g cecal contents. Phage resistance rates of initially up to 43% in the single phage treated group and 24% in the cocktail treated group later stabilized at low levels. The occurrence of phage resistance influenced but did not override the Campylobacter reducing effect. Regarding the reduction potential, the cocktail treatment had only a small advantage over the singe phage treatment directly after phage administration. However, the cocktail moderated and delayed the emergence of phage resistance. PMID:24205254

  13. The in vivo efficacy of two administration routes of a phage cocktail to reduce numbers of Campylobacter coli and Campylobacter jejuni in chickens

    Directory of Open Access Journals (Sweden)

    Carvalho Carla M

    2010-09-01

    Full Text Available Abstract Background Poultry meat is one of the most important sources of human campylobacteriosis, an acute bacterial enteritis which is a major problem worldwide. Campylobacter coli and Campylobacter jejuni are the most common Campylobacter species associated with this disease. These pathogens live in the intestinal tract of most avian species and under commercial conditions they spread rapidly to infect a high proportion of the flock, which makes their treatment and prevention very difficult. Bacteriophages (phages are naturally occurring predators of bacteria with high specificity and also the capacity to evolve to overcome bacterial resistance. Therefore phage therapy is a promising alternative to antibiotics in animal production. This study tested the efficacy of a phage cocktail composed of three phages for the control of poultry infected with C. coli and C. jejuni. Moreover, it evaluated the effectiveness of two routes of phage administration (by oral gavage and in feed in order to provide additional information regarding their future use in a poultry unit. Results The results indicate that experimental colonisation of chicks was successful and that the birds showed no signs of disease even at the highest dose of Campylobacter administered. The phage cocktail was able to reduce the titre of both C. coli and C. jejuni in faeces by approximately 2 log10 cfu/g when administered by oral gavage and in feed. This reduction persisted throughout the experimental period and neither pathogen regained their former numbers. The reduction in Campylobacter titre was achieved earlier (2 days post-phage administration when the phage cocktail was incorporated in the birds' feed. Campylobacter strains resistant to phage infection were recovered from phage-treated chickens at a frequency of 13%. These resistant phenotypes did not exhibit a reduced ability to colonize the chicken guts and did not revert to sensitive types. Conclusions Our findings provide

  14. Phage-displayed peptides selected for binding to Campylobacter jejuni are antimicrobial.

    Science.gov (United States)

    Bishop-Hurley, Sharon L; Rea, Philippa J; McSweeney, Christopher S

    2010-10-01

    In developed countries, Campylobacter jejuni is a leading cause of zoonotic bacterial gastroenteritis in humans with chicken meat implicated as a source of infection. Campylobacter jejuni colonises the lower gastrointestinal tract of poultry and during processing is spread from the gastrointestinal tract onto the surface of dressed carcasses. Controlling or eliminating C.jejuni on-farm is considered to be one of the best strategies for reducing human infection. Molecules on the cell surface of C.jejuni interact with the host to facilitate its colonisation and persistence in the gastrointestinal tract of poultry. We used a subtractive phage-display protocol to affinity select for peptides binding to the cell surface of a poultry isolate of C.jejuni with the aim of finding peptides that could be used to control this microorganism in chickens. In total, 27 phage peptides, representing 11 unique clones, were found to inhibit the growth of C.jejuni by up to 99.9% in vitro. One clone was bactericidal, reducing the viability of C.jejuni by 87% in vitro. The phage peptides were highly specific. They completely inhibited the growth of two of the four poultry isolates of C.jejuni tested with no activity detected towards other Gram-negative and Gram-positive bacteria.

  15. Campylobacters and their bacteriophages from chicken liver: The prospect for phage biocontrol.

    Science.gov (United States)

    Firlieyanti, Antung S; Connerton, Phillippa L; Connerton, Ian F

    2016-11-21

    Consumption of foods containing chicken liver has been associated with Campylobacter enteritis. Campylobacters can contaminate the surface of livers post-mortem but can also arise through systemic infection of colonising bacteria in live birds. The use of bacteriophage to reduce levels of Campylobacter entering the food chain is a promising intervention approach but most phages have been isolated from chicken excreta. This study examined the incidence and contamination levels of Campylobacter and their bacteriophage in UK retail chicken liver. Using enrichment procedures, 87% of 109 chicken livers were surface contaminated with Campylobacter and 83% contaminated within internal tissues. Direct plating on selective agar allowed enumeration of viable bacteria from 43% of liver samples with counts ranging from 1.8->3.8log10CFU/cm(2) for surface samples, and 3.0->3.8log10CFU/g for internal tissue samples. Three C. jejuni isolates recovered from internal liver tissues were assessed for their ability to colonise the intestines and extra-intestinal organs of broiler chickens following oral infection. All isolates efficiently colonised the chicken intestines but were variable in their abilities to colonise extra-intestinal organs. One isolate, CLB104, could be recovered by enrichment from the livers and kidneys of three of seven chickens. Campylobacter isolates remained viable within fresh livers stored at 4°C over 72h and frozen livers stored at -20°C over 7days in atmospheric oxygen, and therefore constitute a risk to human health. Only three Campylobacter-specific bacteriophages were isolated, and these exhibited a limited host range against the Camplylobacter chicken liver isolates. All were identified as group III virulent bacteriophage based on their genome size of 140kb. The application of broad host range group II virulent phages (8log10PFU/g) to liver homogenates containing C. jejuni strains of diverse origin at 4°C resulted in modest but significant reductions

  16. Identification of antigenic epitopes of the SapA protein of Campylobacter fetus using a phage display peptide library.

    Science.gov (United States)

    Zhao, Hailing; Yu, Shenye; Liu, Huifang; Si, Wei; Wang, Chunlai; Liu, Siguo

    2012-12-01

    In this study, we immunized mice with prokaryotically expressed recombinant surface layer protein, SapA, of Campylobacter fetus, generated hybridomas secreting mouse monoclonal antibodies (mAb) targeting SapA, and purified the mAb A2D5 from mouse ascites using saturated ammonium sulfate solution. The mAb A2D5, coated onto ELISA plates, was used to screen the phage random 12-peptide library through three rounds of panning. Following panning, 15 phage clones were randomly chosen and tested for reactivity with mAb A2D5 by indirect ELISA. Single-stranded DNA from positive clones was sequenced and compared with the sequence of SapA to predict the key epitope. ELISA and/or Western blot analyses further validated that synthetic peptides and recombinant peptide mimotopes all interact with mAb A2D5. Nine of ten positive phage clones identified by screening were sequenced successfully. Seven clones shared the same sequence HYDRHNYHWWHT; one had the sequence LSKNLPLTALGN; and the final one had the sequence SGMKEPELRSYS. These three sequences shared high homology with SapA J05577 in the region GNEKDFVTKIYSIALGNTSDVDGINYW, in which the underlined amino acids may serve as key residues in the epitope. ELISA and/or Western blot analyses showed that mAb A2D5 not only interacted with the four synthetic peptide mimotopes, but also with 14 prokaryotically expressed recombinant peptide mimotopes. The mimotopes identified in this study will aid future studies into the pathological processes and immune mechanisms of the SapA protein of C. fetus. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. Designing phage therapeutics.

    Science.gov (United States)

    Goodridge, Lawrence D

    2010-01-01

    Phage therapy is the application of phages to bodies, substances, or environments to effect the biocontrol of pathogenic or nuisance bacteria. To be effective, phages, minimally, must be capable of attaching to bacteria (adsorption), killing those bacteria (usually associated with phage infection), and otherwise surviving (resisting decay) until they achieve attachment and subsequent killing. While a strength of phage therapy is that phages that possess appropriate properties can be chosen from a large diversity of naturally occurring phages, a more rational approach to phage therapy also can include post-isolation manipulation of phages genetically, phenotypically, or in terms of combining different products into a single formulation. Genetic manipulation, especially in these modern times, can involve genetic engineering, though a more traditional approach involves the selection of spontaneously occurring phage mutants during serial transfer protocols. While genetic modification typically is done to give rise to phenotypic changes in phages, phage phenotype alone can also be modified in vitro, prior to phage application for therapeutic purposes, as for the sake of improving phage lethality (such as by linking phage virions to antibacterial chemicals such as chloramphenicol) or survival capabilities (e.g., via virion PEGylation). Finally, phages, both naturally occurring isolates or otherwise modified constructs, can be combined into cocktails which provide collectively enhanced capabilities such as expanded overall host range. Generally these strategies represent different routes towards improving phage therapy formulations and thereby efficacy through informed design.

  18. Phage therapy pharmacology: calculating phage dosing.

    Science.gov (United States)

    Abedon, Stephen

    2011-01-01

    Phage therapy, which can be described as a phage-mediated biocontrol of bacteria (or, simply, biocontrol), is the application of bacterial viruses-also bacteriophages or phages-to reduce densities of nuisance or pathogenic bacteria. Predictive calculations for phage therapy dosing should be useful toward rational development of therapeutic as well as biocontrol products. Here, I consider the theoretical basis of a number of concepts relevant to phage dosing for phage therapy including minimum inhibitory concentration (but also "inundation threshold"), minimum bactericidal concentration (but also "clearance threshold"), decimal reduction time (D value), time until bacterial eradication, threshold bacterial density necessary to support phage population growth ("proliferation threshold"), and bacterial density supporting half-maximal phage population growth rates (K(B)). I also address the concepts of phage killing titers, multiplicity of infection, and phage peak densities. Though many of the presented ideas are not unique to this chapter, I nonetheless provide variations on derivations and resulting formulae, plus as appropriate discuss relative importance. The overriding goal is to present a variety of calculations that are useful toward phage therapy dosing so that they may be found in one location and presented in a manner that allows facile appreciation, comparison, and implementation. The importance of phage density as a key determinant of the phage potential to eradicate bacterial targets is stressed throughout the chapter.

  19. Complete genome sequence of UV-resistant Campylobacter jejuni RM3194, including an 81.08-kilobase plasmid

    Science.gov (United States)

    Campylobacter jejuni strain RM3194 was originally isolated from a human with enteritis and contains a novel 81,079-bp plasmid. RM3194 has exhibited superior survival compared to other Campylobacter jejuni strains when challenged with UV light. The chromosome of RM3194 was determined to be 1,651,18...

  20. Emended description of Campylobacter sputorum and revision of its infrasubspecific (biovar) divisions, including C-sputorum biovar paraureolyticus, a urease-producing variant from cattle and humans

    DEFF Research Database (Denmark)

    On, S.L.W.; Atabay, H.I.; Corry, J.E.L.;

    1998-01-01

    A polyphasic taxonomic study of 15 bovine and human strains assigned to the catalase-negative, urease-positive campylobacter (CNUPC) group identified these bacteria as a novel, ureolytic biovar of Campylobacter sputorum for which we propose the name C. sputorum bv. paraureolyticus: suitable...... should be revised to include by. sputorum for catalase-negative strains; by. fecalis for catalase-positive strains; and by. paraureolyticus for urease-positive strains. Strains classified previously as by. bubulus should be reclassified as by. sputorum. The species description of C. sputorum is revised...

  1. Sequence variability of Campylobacter temperate bacteriophages

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    Ng Lai-King

    2008-03-01

    Full Text Available Abstract Background Prophages integrated within the chromosomes of Campylobacter jejuni isolates have been demonstrated very recently. Prior work with Campylobacter temperate bacteriophages, as well as evidence from prophages in other enteric bacteria, suggests these prophages might have a role in the biology and virulence of the organism. However, very little is known about the genetic variability of Campylobacter prophages which, if present, could lead to differential phenotypes in isolates carrying the phages versus those that do not. As a first step in the characterization of C. jejuni prophages, we investigated the distribution of prophage DNA within a C. jejuni population assessed the DNA and protein sequence variability within a subset of the putative prophages found. Results Southern blotting of C. jejuni DNA using probes from genes within the three putative prophages of the C. jejuni sequenced strain RM 1221 demonstrated the presence of at least one prophage gene in a large proportion (27/35 of isolates tested. Of these, 15 were positive for 5 or more of the 7 Campylobacter Mu-like phage 1 (CMLP 1, also designated Campylobacter jejuni integrated element 1, or CJIE 1 genes tested. Twelve of these putative prophages were chosen for further analysis. DNA sequencing of a 9,000 to 11,000 nucleotide region of each prophage demonstrated a close homology with CMLP 1 in both gene order and nucleotide sequence. Structural and sequence variability, including short insertions, deletions, and allele replacements, were found within the prophage genomes, some of which would alter the protein products of the ORFs involved. No insertions of novel genes were detected within the sequenced regions. The 12 prophages and RM 1221 had a % G+C very similar to C. jejuni sequenced strains, as well as promoter regions characteristic of C. jejuni. None of the putative prophages were successfully induced and propagated, so it is not known if they were functional or

  2. Genomic comparison of Escherichia coli O104:H4 isolates from 2009 and 2011 reveals plasmid, and prophage heterogeneity, including shiga toxin encoding phage stx2.

    Directory of Open Access Journals (Sweden)

    Sanaa A Ahmed

    Full Text Available In May of 2011, an enteroaggregative Escherichia coli O104:H4 strain that had acquired a Shiga toxin 2-converting phage caused a large outbreak of bloody diarrhea in Europe which was notable for its high prevalence of hemolytic uremic syndrome cases. Several studies have described the genomic inventory and phylogenies of strains associated with the outbreak and a collection of historical E. coli O104:H4 isolates using draft genome assemblies. We present the complete, closed genome sequences of an isolate from the 2011 outbreak (2011C-3493 and two isolates from cases of bloody diarrhea that occurred in the Republic of Georgia in 2009 (2009EL-2050 and 2009EL-2071. Comparative genome analysis indicates that, while the Georgian strains are the nearest neighbors to the 2011 outbreak isolates sequenced to date, structural and nucleotide-level differences are evident in the Stx2 phage genomes, the mer/tet antibiotic resistance island, and in the prophage and plasmid profiles of the strains, including a previously undescribed plasmid with homology to the pMT virulence plasmid of Yersinia pestis. In addition, multiphenotype analysis showed that 2009EL-2071 possessed higher resistance to polymyxin and membrane-disrupting agents. Finally, we show evidence by electron microscopy of the presence of a common phage morphotype among the European and Georgian strains and a second phage morphotype among the Georgian strains. The presence of at least two stx2 phage genotypes in host genetic backgrounds that may derive from a recent common ancestor of the 2011 outbreak isolates indicates that the emergence of stx2 phage-containing E. coli O104:H4 strains probably occurred more than once, or that the current outbreak isolates may be the result of a recent transfer of a new stx2 phage element into a pre-existing stx2-positive genetic background.

  3. Engineered phages for electronics.

    Science.gov (United States)

    Cui, Yue

    2016-11-15

    Phages are traditionally widely studied in biology and chemistry. In recent years, engineered phages have attracted significant attentions for functionalization or construction of electronic devices, due to their specific binding, catalytic, nucleating or electronic properties. To apply the engineered phages in electronics, these are a number of interesting questions: how to engineer phages for electronics? How are the engineered phages characterized? How to assemble materials with engineered phages? How are the engineered phages micro or nanopatterned? What are the strategies to construct electronics devices with engineered phages? This review will highlight the early attempts to address these questions and explore the fundamental and practical aspects of engineered phages in electronics, including the approaches for selection or expression of specific peptides on phage coat proteins, characterization of engineered phages in electronics, assembly of electronic materials, patterning of engineered phages, and construction of electronic devices. It provides the methodologies and opens up ex-cit-ing op-por-tu-ni-ties for the development of a variety of new electronic materials and devices based on engineered phages for future applications.

  4. A longitudinal study of Campylobacter distribution in a turkey production Chain in Finland

    DEFF Research Database (Denmark)

    Perko-Makela, P.; Isohanni, P.; Katzav, M.

    2007-01-01

    Campylobacter is a leading course of food borne bacterial infections worldwide. It is believed that a decline in the occurrence of campylobacteriosis can be achieved by reducing the number of the bacterium in the gastrointestinal tract of poultry. Phage therapy using Campylobacter specific phages...... given to poultry prior to slaughter is a promising control measure. However, the reducing effect of most phages tested so far is rather limited due to the development of phage resistant Campylobacter. An increased knowledge of the molecular mechanisms of importance for development of phage resistant...... Campylobacter is therefore essential for implementation of this method. Recently, a collection of Campylobacter bacteriophages has been established. The phages were isolated from faeces from broilers and ducks and it was shown by transmission electron microscopy that they all belong to the family of Myoviridae...

  5. Methods for initial characterization of Campylobacter jejuni bacteriophages

    DEFF Research Database (Denmark)

    Sørensen, Martine Camilla Holst; Gencay, Yilmaz Emre; Brøndsted, Lone

    2017-01-01

    Here we describe an initial characterization of Campylobacter jejuni bacteriophages by host range analysis, genome size determination by pulsed-field gel electrophoresis, and receptor-type identification by screening mutants for phage sensitivity.......Here we describe an initial characterization of Campylobacter jejuni bacteriophages by host range analysis, genome size determination by pulsed-field gel electrophoresis, and receptor-type identification by screening mutants for phage sensitivity....

  6. Alternative bacteriophage life cycles: the carrier state of Campylobacter jejuni.

    Science.gov (United States)

    Siringan, Patcharin; Connerton, Phillippa L; Cummings, Nicola J; Connerton, Ian F

    2014-03-26

    Members of the genus Campylobacter are frequently responsible for human enteric disease, often through consumption of contaminated poultry products. Bacteriophages are viruses that have the potential to control pathogenic bacteria, but understanding their complex life cycles is key to their successful exploitation. Treatment of Campylobacter jejuni biofilms with bacteriophages led to the discovery that phages had established a relationship with their hosts typical of the carrier state life cycle (CSLC), where bacteria and bacteriophages remain associated in equilibrium. Significant phenotypic changes include improved aerotolerance under nutrient-limited conditions that would confer an advantage to survive in extra-intestinal environments, but a lack in motility eliminated their ability to colonize chickens. Under these circumstances, phages can remain associated with a compatible host and continue to produce free virions to prospect for new hosts. Moreover, we demonstrate that CSLC host bacteria can act as expendable vehicles for the delivery of bacteriophages to new host bacteria within pre-colonized chickens. The CSLC represents an important phase in the ecology of Campylobacter bacteriophage.

  7. Campylobacter infection

    Science.gov (United States)

    ... in the small intestine from a bacteria called Campylobacter jejuni . It is a type of food poisoning. Causes ... testing for white blood cells Stool culture for Campylobacter jejuni Treatment The infection almost always goes away on ...

  8. Campylobacter Infections

    Science.gov (United States)

    Campylobacter infection is a common foodborne illness. You get it from eating raw or undercooked poultry. You ... whether you need to take antibiotics. To prevent campylobacter infection, cook poultry thoroughly. Use a separate cutting ...

  9. Phage therapy pharmacology phage cocktails.

    Science.gov (United States)

    Chan, Benjamin K; Abedon, Stephen T

    2012-01-01

    Phage therapy is the clinical or veterinary application of bacterial viruses (bacteriophages) as antibacterial "drugs." More generally, phages can be used as biocontrol agents against plant as well as foodborne pathogens. In this chapter, we consider the therapeutic use of phage cocktails, which is the combining of two or more phage types to produce more pharmacologically diverse formulations. The primary motivation for the use of cocktails is their broader spectra of activity in comparison to individual phage isolates: they can impact either more bacterial types or achieve effectiveness under a greater diversity of conditions. The combining of phages can also facilitate better targeting of multiple strains making up individual bacterial species or covering multiple species that might be responsible for similar disease states, in general providing, relative to individual phage isolates, a greater potential for presumptive or empirical treatment. Contrasting the use of phage banks, or even phage isolation against specific etiologies that have been obtained directly from patients under treatment, here we consider the utility as well as potential shortcomings associated with the use of phage cocktails as therapeutic antibacterial agents.

  10. Effect of bacteriophage application on Campylobacter jejuni loads in commercial broiler flocks.

    Science.gov (United States)

    Kittler, Sophie; Fischer, Samuel; Abdulmawjood, Amir; Glünder, Gerhard; Klein, Günter

    2013-12-01

    Campylobacteriosis is the most frequent food-borne human enteritis. The major source for infection with Campylobacter spp. is broiler meat. Risk assessments consider the reduction of Campylobacter in primary production to be most beneficial for human health. The aim of this study was to test the efficacy of a bacteriophage application under commercial conditions which had proved to be effective in previous noncommercial studies under controlled experimental conditions. A phage cocktail for Campylobacter reduction was tested on three commercial broiler farms each with a control and an experimental group. Colonization of Campylobacter was confirmed prior to phage application in fecal samples. Subsequently, a phage cocktail was applied via drinking water in the experimental group (log10 5.8 to 7.5 PFU/bird). One day after phage application, Campylobacter counts of one experimental group were reduced under the detection limit (log10 3.2 CFU/g cecal content compared to the control was still detected (P=0.0011). No significant reduction was observed in the experimental groups of the other trials. However, a significant drop in cecal Campylobacter counts occurred in a phage-contaminated control. These results suggest that maximum reduction of Campylobacter at the slaughterhouse might be achieved by phage application 1 to 4 days prior to slaughter.

  11. Bacteriophages to combat foodborne infections caused by food contamination by bacteria of the Campylobacter genus

    Directory of Open Access Journals (Sweden)

    Magdalena Myga-Nowak

    2016-09-01

    Full Text Available It is estimated that each year more than 2 million people suffer from diarrheal diseases, resulting from the consumption of contaminated meat. Foodborne infections are most frequently caused by small Gram-negative rods Campylobacter. The hosts of these bacteria are mainly birds wherein they are part of the normal intestinal flora. During the commercial slaughter, there is a likelihood of contamination of carcasses by the bacteria found in the intestinal content. In Europe, up to 90% of poultry flocks can be a reservoir of the pathogen. According to the European Food Safety Authority report from 2015, the number of reported and confirmed cases of human campylobacteriosis exceeds 200 thousands per year, and such trend remains at constant level for several years. The occurrence of growing antibiotic resistance in bacteria forces the limitation of antibiotic use in the animal production. Therefore, the European Union allows only using stringent preventive and hygienic treatment on farms. Achieving Campylobacter free chickens using these methods is possible, but difficult to implement and expensive. Utilization of bacterial viruses – bacteriophages, can be a path to provide the hygienic conditions of poultry production and food processing. Formulations applied in the food protection should contain strictly lytic bacteriophages, be non-pyrogenic and retain long lasting biological activity. Currently, on the market there are available commercial bacteriophage preparations for agricultural use, but neither includes phages against Campylobacter. However, papers on the application of bacteriophages against Campylobacter in chickens and poultry products were published in the last few years. In accordance with the estimates, 2-logarithm reduction of Campylobacter in poultry carcases will contribute to the 30-fold reduction in the incidence of campylobacteriosis in humans. Research on bacteriophages against Campylobacter have cognitive and economic

  12. Clostridium difficile phages: still difficult?

    Directory of Open Access Journals (Sweden)

    Katherine Rose Hargreaves

    2014-04-01

    Full Text Available Phages that infect Clostridium difficile were first isolated for typing purposes in the 1980s, but their use was short lived. However, the rise of C. difficile epidemics over the last decade has triggered a resurgence of interest in using phages to combat this pathogen. Phage therapy is an attractive treatment option for C. difficile infection, however developing suitable phages is challenging. In this review we summarise the difficulties faced by researchers in this field, and we discuss the solutions and strategies used for the development of C. difficile phages for use as novel therapeutics.Epidemiological data has highlighted the diversity and distribution of C. difficile, and shown that novel strains continue to emerge in clinical settings. In parallel with epidemiological studies, advances in molecular biology have bolstered our understanding of C. difficile biology, and our knowledge of phage-host interactions in other bacterial species. These three fields of biology have therefore paved the way for future work on C. difficile phages to progress and develop. Benefits of using C. difficile phages as therapeutic agents include the fact that they have highly specific interactions with their bacterial hosts. Studies also show that they can reduce bacterial numbers in both in vitro and in vivo systems. Genetic analysis has revealed the genomic diversity among these phages and provided an insight into their taxonomy and evolution.No strictly virulent C. difficile phages have been reported and this contributes to the difficulties with their therapeutic exploitation. Although treatment approaches using the phage-encoded endolysin protein have been explored, the benefits of using whole-phages are such that they remain a major research focus. Whilst we don’t envisage working with C. difficile phages will be problem free, sufficient study should inform future strategies to facilitate their development to combat this problematic pathogen.

  13. Methods for Isolation, Purification, and Propagation of Bacteriophages of Campylobacter jejuni

    DEFF Research Database (Denmark)

    Gencay, Yilmaz Emre; Birk, Tina; Sørensen, Martine Camilla Holst

    2017-01-01

    Here, we describe the methods for isolation, purification, and propagation of Campylobacter jejuni bacteriophages from samples expected to contain high number of phages such as chicken feces. The overall steps are (1) liberation of phages from the sample material; (2) observation of plaque...

  14. Campylobacter serology test

    Science.gov (United States)

    ... time the skin is broken) Images Blood test Campylobacter jejuni organism References Allos BM. Campylobacter infections. In: Goldman ... chap 303. Allos BM, Iovine NM, Blaser MJ. Campylobacter jejuni and related species. In: Bennett JE, Dolin R, ...

  15. Molecular and Statistical Analysis of Campylobacter spp. and Antimicrobial-Resistant Campylobacter Carriage in Wildlife and Livestock from Ontario Farms.

    Science.gov (United States)

    Viswanathan, M; Pearl, D L; Taboada, E N; Parmley, E J; Mutschall, S; Jardine, C M

    2017-05-01

    The objectives of this study were to (i) compare the carriage of Campylobacter and antimicrobial-resistant Campylobacter among livestock and mammalian wildlife on Ontario farms, and (ii) investigate the potential sharing of Campylobacter subtypes between livestock and wildlife. Using data collected from a cross-sectional study of 25 farms in 2010, we assessed associations, using mixed logistic regression models, between Campylobacter and antimicrobial-resistant Campylobacter carriage and the following explanatory variables: animal species (beef, dairy, swine, raccoon, other), farm type (swine, beef, dairy), type of sample (livestock or wildlife) and Campylobacter species (jejuni, coli, other). Models included a random effect to account for clustering by farm where samples were collected. Samples were subtyped using a Campylobacter-specific 40 gene comparative fingerprinting assay. A total of 92 livestock and 107 wildlife faecal samples were collected, and 72% and 27% tested positive for Campylobacter, respectively. Pooled faecal samples from livestock were significantly more likely to test positive for Campylobacter than wildlife samples. Relative to dairy cattle, pig samples were at significantly increased odds of testing positive for Campylobacter. The odds of isolating Campylobacter jejuni from beef cattle samples were significantly greater compared to dairy cattle and raccoon samples. Fifty unique subtypes of Campylobacter were identified, and only one subtype was found in both wildlife and livestock samples. Livestock Campylobacter isolates were significantly more likely to exhibit antimicrobial resistance (AMR) compared to wildlife Campylobacter isolates. Campylobacter jejuni was more likely to exhibit AMR when compared to C. coli. However, C. jejuni isolates were only resistant to tetracycline, and C.  coli isolates exhibited multidrug resistance patterns. Based on differences in prevalence of Campylobacter spp. and resistant Campylobacter between

  16. Phage display-derived binders able to distinguish Listeria monocytogenes from other Listeria species.

    Directory of Open Access Journals (Sweden)

    Josephine Morton

    Full Text Available The objective of this study was to produce phage display-derived binders with the ability to distinguish Listeria monocytogenes from other Listeria spp., which may have potential utility to enhance detection of Listeria monocytogenes. To obtain binders with the desired binding specificity a series of surface and solution phage-display biopannings were performed. Initially, three rounds of surface biopanning against gamma-irradiated L. monocytogenes serovar 4b cells were performed followed by an additional surface biopanning round against L. monocytogenes 4b which included prior subtraction biopanning against gamma-irradiated L. innocua cells. In an attempt to further enhance binder specificity for L. monocytogenes 4b two rounds of solution biopanning were performed, both rounds included initial subtraction solution biopanning against L. innocua. Subsequent evaluations were performed on the phage clones by phage binding ELISA. All phage clones tested from the second round of solution biopanning had higher specificity for L. monocytogenes 4b than for L. innocua and three other foodborne pathogens (Salmonella spp., Escherichia coli and Campylobacter jejuni. Further evaluation with five other Listeria spp. revealed that one phage clone in particular, expressing peptide GRIADLPPLKPN, was highly specific for L. monocytogenes with at least 43-fold more binding capability to L. monocytogenes 4b than to any other Listeria sp. This proof-of-principle study demonstrates how a combination of surface, solution and subtractive biopanning was used to maximise binder specificity. L. monocytogenes-specific binders were obtained which could have potential application in novel detection tests for L. monocytogenes, benefiting both the food and medical industries.

  17. Phage display-derived binders able to distinguish Listeria monocytogenes from other Listeria species.

    Science.gov (United States)

    Morton, Josephine; Karoonuthaisiri, Nitsara; Charlermroj, Ratthaphol; Stewart, Linda D; Elliott, Christopher T; Grant, Irene R

    2013-01-01

    The objective of this study was to produce phage display-derived binders with the ability to distinguish Listeria monocytogenes from other Listeria spp., which may have potential utility to enhance detection of Listeria monocytogenes. To obtain binders with the desired binding specificity a series of surface and solution phage-display biopannings were performed. Initially, three rounds of surface biopanning against gamma-irradiated L. monocytogenes serovar 4b cells were performed followed by an additional surface biopanning round against L. monocytogenes 4b which included prior subtraction biopanning against gamma-irradiated L. innocua cells. In an attempt to further enhance binder specificity for L. monocytogenes 4b two rounds of solution biopanning were performed, both rounds included initial subtraction solution biopanning against L. innocua. Subsequent evaluations were performed on the phage clones by phage binding ELISA. All phage clones tested from the second round of solution biopanning had higher specificity for L. monocytogenes 4b than for L. innocua and three other foodborne pathogens (Salmonella spp., Escherichia coli and Campylobacter jejuni). Further evaluation with five other Listeria spp. revealed that one phage clone in particular, expressing peptide GRIADLPPLKPN, was highly specific for L. monocytogenes with at least 43-fold more binding capability to L. monocytogenes 4b than to any other Listeria sp. This proof-of-principle study demonstrates how a combination of surface, solution and subtractive biopanning was used to maximise binder specificity. L. monocytogenes-specific binders were obtained which could have potential application in novel detection tests for L. monocytogenes, benefiting both the food and medical industries.

  18. Campylobacter bacteriophages and bacteriophage therapy.

    Science.gov (United States)

    Connerton, P L; Timms, A R; Connerton, I F

    2011-08-01

    Members of the genus Campylobacter are frequently responsible for human enteric disease with occasionally very serious outcomes. Much of this disease burden is thought to arise from consumption of contaminated poultry products. More than 80% of poultry in the UK harbour Campylobacter as a part of their intestinal flora. To address this unacceptably high prevalence, various interventions have been suggested and evaluated. Among these is the novel approach of using Campylobacter-specific bacteriophages, which are natural predators of the pathogen. To optimize their use as therapeutic agents, it is important to have a comprehensive understanding of the bacteriophages that infect Campylobacter, and how they can affect their host bacteria. This review will focus on many aspects of Campylobacter-specific bacteriophages including: their first isolation in the 1960s, their use in bacteriophage typing schemes, their isolation from the different biological sources and genomic characterization. As well as their use as therapeutic agents to reduce Campylobacter in poultry their future potential, including their use in bio-sanitization of food, will be explored. The evolutionary consequences of naturally occurring bacteriophage infection that have come to light through investigations of bacteriophages in the poultry ecosystem will also be discussed.

  19. Integrated approach leads to less Campylobacter

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Hald, Birthe; Borck Høg, Birgitte

    2012-01-01

    Methods of reducing the risk of Campylobacter infection during indoor broiler (chicken) production are discussed, including: risk management intiatives; biosecurity measures; scheduled slaughter; hygiene and decontamination; and improving consumer information.......Methods of reducing the risk of Campylobacter infection during indoor broiler (chicken) production are discussed, including: risk management intiatives; biosecurity measures; scheduled slaughter; hygiene and decontamination; and improving consumer information....

  20. Post-genome Analysis of the Foodborne Pathogen Campylobacter jejuni

    Science.gov (United States)

    Kay, Emily J.; Gundogdu, Ozan; Wren, Brendan

    The human pathogen Campylobacter jejuni is part of the genus Campylobacter that lies within the epsilon proteobacteria subclass of bacteria. The nearest family in phylogenetic terms is the Helicobacteraceae which includes the Helicobacter and Wolinella genuses. Campylobacter species are Gram-negative, curved rod shaped or spiral and are motile (via polar flagella).

  1. Two flagellotropic phages and one pilus-specific phage active against Asticcacaulis biprosthecum.

    Science.gov (United States)

    Pate, J L; Petzold, S J; Umbreit, T H

    1979-04-15

    Three phages active against cells of Asticcacaulis biprosthecum attach to receptor sites located at the pole of the cell where pili, flagella, and holdfast are produced. Phage phiAcS2, a large phage with a prolate cylindrical head and flexible, noncontractile tail, attaches to flagella as well as to receptor sites at the pole of the cell. Attachment to flagella occurs at the region where head and tail of the phage are joined, leaving the distal end of the tail free for attachment to receptor sites at the cell surface. Phages phiAcM2 and phiAcM4, are identical in appearance to each other, possessing prolate cylindrical heads and flexible, noncontractile tails, and are smaller than phage phiAcS2. Phage phiAcM4, exhibits the same flagellotropic characteristic as described for phage phiAcS2, including the manner of attachment to flagella. Phage phiAcM2 has no affinity for flagella, but attaches by the distal end of the tail to pili and to receptor sites at the pole of the cell. Mechanical removal of flagella and pili protects against infection by all three phages. Studies with phage-resistant mutants and with KCN-treated cells suggest that pili are required for infection by both flagellotropic and pilus-specific phages.

  2. Global Epidemiology of Campylobacter Infection

    Science.gov (United States)

    Kaakoush, Nadeem O.; Castaño-Rodríguez, Natalia; Mitchell, Hazel M.

    2015-01-01

    SUMMARY Campylobacter jejuni infection is one of the most widespread infectious diseases of the last century. The incidence and prevalence of campylobacteriosis have increased in both developed and developing countries over the last 10 years. The dramatic increase in North America, Europe, and Australia is alarming, and data from parts of Africa, Asia, and the Middle East indicate that campylobacteriosis is endemic in these areas, especially in children. In addition to C. jejuni, there is increasing recognition of the clinical importance of emerging Campylobacter species, including Campylobacter concisus and Campylobacter ureolyticus. Poultry is a major reservoir and source of transmission of campylobacteriosis to humans. Other risk factors include consumption of animal products and water, contact with animals, and international travel. Strategic implementation of multifaceted biocontrol measures to reduce the transmission of this group of pathogens is paramount for public health. Overall, campylobacteriosis is still one of the most important infectious diseases that is likely to challenge global health in the years to come. This review provides a comprehensive overview of the global epidemiology, transmission, and clinical relevance of Campylobacter infection. PMID:26062576

  3. Recombinant phage probes for Listeria monocytogenes

    Energy Technology Data Exchange (ETDEWEB)

    Carnazza, S; Gioffre, G; Felici, F; Guglielmino, S [Department of Microbiological, Genetic and Molecular Sciences, University of Messina, Messina (Italy)

    2007-10-03

    Monitoring of food and environmental samples for biological threats, such as Listeria monocytogenes, requires probes that specifically bind biological agents and ensure their immediate and efficient detection. There is a need for robust and inexpensive affinity probes as an alternative to antibodies. These probes may be recruited from random peptide libraries displayed on filamentous phage. In this study, we selected from two phage peptide libraries phage clones displaying peptides capable of specific and strong binding to the L. monocytogenes cell surface. The ability of isolated phage clones to interact specifically with L. monocytogenes was demonstrated using enzyme-linked immunosorbent assay (ELISA) and confirmed by co-precipitation assay. We also assessed the sensitivity of phage-bacteria binding by PCR on phage-captured Listeria cells, which could be detected at a concentration of 10{sup 4} cells ml{sup -1}. In addition, as proof-of-concept, we tested the possibility of immobilizing the affinity-selected phages to a putative biosensor surface. The quality of phage deposition was monitored by ELISA and fluorescent microscopy. Phage-bacterial binding was confirmed by high power optical phase contrast microscopy. Overall, the results of this work validate the concept of affinity-selected recombinant filamentous phages as probes for detecting and monitoring bacterial agents under any conditions that warrant their recognition, including in food products.

  4. Recombinant phage probes for Listeria monocytogenes

    Science.gov (United States)

    Carnazza, S.; Gioffrè, G.; Felici, F.; Guglielmino, S.

    2007-10-01

    Monitoring of food and environmental samples for biological threats, such as Listeria monocytogenes, requires probes that specifically bind biological agents and ensure their immediate and efficient detection. There is a need for robust and inexpensive affinity probes as an alternative to antibodies. These probes may be recruited from random peptide libraries displayed on filamentous phage. In this study, we selected from two phage peptide libraries phage clones displaying peptides capable of specific and strong binding to the L. monocytogenes cell surface. The ability of isolated phage clones to interact specifically with L. monocytogenes was demonstrated using enzyme-linked immunosorbent assay (ELISA) and confirmed by co-precipitation assay. We also assessed the sensitivity of phage-bacteria binding by PCR on phage-captured Listeria cells, which could be detected at a concentration of 104 cells ml-1. In addition, as proof-of-concept, we tested the possibility of immobilizing the affinity-selected phages to a putative biosensor surface. The quality of phage deposition was monitored by ELISA and fluorescent microscopy. Phage-bacterial binding was confirmed by high power optical phase contrast microscopy. Overall, the results of this work validate the concept of affinity-selected recombinant filamentous phages as probes for detecting and monitoring bacterial agents under any conditions that warrant their recognition, including in food products.

  5. Campylobacter jejuni organism (image)

    Science.gov (United States)

    Campylobacter jejuni infection causes cramping, diarrhea, abdominal pain and fever within 2 to 5 days after a person has been exposed to the organism. Campylobacter jejuni is one of the most common bacterial causes ...

  6. Use of culture, PCR analysis, and DNA microarrays for detection of Campylobacter jejuni and Campylobacter coli from chicken feces

    DEFF Research Database (Denmark)

    Keramas, Georgios; Bang, Dang Duong; Lund, Marianne

    2004-01-01

    A DNA microarray for detection of Campylobacter spp. was recently developed and applied to detect Campylobacter spp. directly from chicken feces. Sixty-five pooled chicken cloacal swab samples from 650 individual broiler chickens were included in the study. The results of Campylobacter sp....... detection obtained with DNA microarrays were compared to those obtained by conventional culture and gel electrophoresis. By conventional culture, 60% of the samples were positive for either Campylobacter jejuni or Campylobacter coli. By PCR and capillary electrophoresis, 95% of the samples were positive...... for Campylobacter spp., whereas with DNA microarrays all samples were positive for Campylobacter spp. By application of DNA microarray analysis, the isolates in 4 samples (6%) could not be identified to the species level, whereas by PCR-capillary electrophoresis, the isolates in 12 samples (19%) remained...

  7. The genome and proteome of a Campylobacter coli bacteriophage vB_CcoM-IBB_35 reveal unusual features

    Directory of Open Access Journals (Sweden)

    Carvalho Carla M

    2012-01-01

    Full Text Available Abstract Background Campylobacter is the leading cause of foodborne diseases worldwide. Bacteriophages (phages are naturally occurring predators of bacteria, ubiquitous in the environment, with high host specificity and thus considered an appealing option to control bacterial pathogens. Nevertheless for an effective use of phages as antimicrobial agents, it is important to understand phage biology which renders crucial the analysis of phage genomes and proteomes. The lack of sequence data from Campylobacter phages adds further importance to these studies. Methods vB_CcoM-IBB_35 is a broad lytic spectrum Myoviridae Campylobacter phage with high potential for therapeutic use. The genome of this phage was obtained by pyrosequencing and the sequence data was further analyzed. The proteomic analysis was performed by SDS-PAGE and Mass spectrometry. Results and conclusions The DNA sequence data of vB_CcoM-IBB_35 consists of five contigs for a total of 172,065 bp with an average GC content of 27%. Attempts to close the gaps between contigs were unsuccessful since the DNA preparations appear to contain substances that inhibited Taq and ϕ29 polymerases. From the 210 identified ORFs, around 60% represent proteins that were not functionally assigned. Homology exists with members of the Teequatrovirinae namely for T4 proteins involved in morphogenesis, nucleotide metabolism, transcription, DNA replication and recombination. Tandem mass spectrometric analysis revealed 38 structural proteins as part of the mature phage particle. Conclusions Genes encoding proteins involved in the carbohydrate metabolism along with several incidences of gene duplications, split genes with inteins and introns have been rarely found in other phage genomes yet are found in this phage. We identified the genes encoding for tail fibres and for the lytic cassette, this later, expressing enzymes for bacterial capsular polysaccharides (CPS degradation, which has not been reported

  8. Campylobacter species in animal, food, and environmental sources, and relevant testing programs in Canada.

    Science.gov (United States)

    Huang, Hongsheng; Brooks, Brian W; Lowman, Ruff; Carrillo, Catherine D

    2015-10-01

    Campylobacter species, particularly thermophilic campylobacters, have emerged as a leading cause of human foodborne gastroenteritis worldwide, with Campylobacter jejuni, Campylobacter coli, and Campylobacter lari responsible for the majority of human infections. Although most cases of campylobacteriosis are self-limiting, campylobacteriosis represents a significant public health burden. Human illness caused by infection with campylobacters has been reported across Canada since the early 1970s. Many studies have shown that dietary sources, including food, particularly raw poultry and other meat products, raw milk, and contaminated water, have contributed to outbreaks of campylobacteriosis in Canada. Campylobacter spp. have also been detected in a wide range of animal and environmental sources, including water, in Canada. The purpose of this article is to review (i) the prevalence of Campylobacter spp. in animals, food, and the environment, and (ii) the relevant testing programs in Canada with a focus on the potential links between campylobacters and human health in Canada.

  9. Quantitative models of in vitro bacteriophage-host dynamics and their application to phage therapy.

    Directory of Open Access Journals (Sweden)

    Benjamin J Cairns

    2009-01-01

    Full Text Available Phage therapy is the use of bacteriophages as antimicrobial agents for the control of pathogenic and other problem bacteria. It has previously been argued that successful application of phage therapy requires a good understanding of the non-linear kinetics of phage-bacteria interactions. Here we combine experimental and modelling approaches to make a detailed examination of such kinetics for the important food-borne pathogen Campylobacter jejuni and a suitable virulent phage in an in vitro system. Phage-insensitive populations of C. jejuni arise readily, and as far as we are aware this is the first phage therapy study to test, against in vitro data, models for phage-bacteria interactions incorporating phage-insensitive or resistant bacteria. We find that even an apparently simplistic model fits the data surprisingly well, and we confirm that the so-called inundation and proliferation thresholds are likely to be of considerable practical importance to phage therapy. We fit the model to time series data in order to estimate thresholds and rate constants directly. A comparison of the fit for each culture reveals density-dependent features of phage infectivity that are worthy of further investigation. Our results illustrate how insight from empirical studies can be greatly enhanced by the use of kinetic models: such combined studies of in vitro systems are likely to be an essential precursor to building a meaningful picture of the kinetic properties of in vivo phage therapy.

  10. [Peptide phage display in biotechnology and biomedicine].

    Science.gov (United States)

    Kuzmicheva, G A; Belyavskaya, V A

    2016-07-01

    To date peptide phage display is one of the most common combinatorial methods used for identifying specific peptide ligands. Phage display peptide libraries containing billions different clones successfully used for selection of ligands with high affinity and selectivity toward wide range of targets including individual proteins, bacteria, viruses, spores, different kind of cancer cells and variety of nonorganic targets (metals, alloys, semiconductors etc.) Success of using filamentous phage in phage display technologies relays on the robustness of phage particles and a possibility to genetically modify its DNA to construct new phage variants with novel properties. In this review we are discussing characteristics of the most known non-commercial peptide phage display libraries of different formats (landscape libraries in particular) and their successful applications in several fields of biotechnology and biomedicine: discovery of peptides with diagnostic values against different pathogens, discovery and using of peptides recognizing cancer cells, trends in using of phage display technologies in human interactome studies, application of phage display technologies in construction of novel nano materials.

  11. Integrated approach leads to less Campylobacter

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Hald, Birthe; Borck Høg, Birgitte

    2012-01-01

    Methods of reducing the risk of Campylobacter infection during indoor broiler (chicken) production are discussed, including: risk management intiatives; biosecurity measures; scheduled slaughter; hygiene and decontamination; and improving consumer information....

  12. Clinical aspects of phage therapy.

    Science.gov (United States)

    Międzybrodzki, Ryszard; Borysowski, Jan; Weber-Dąbrowska, Beata; Fortuna, Wojciech; Letkiewicz, Sławomir; Szufnarowski, Krzysztof; Pawełczyk, Zdzisław; Rogóż, Paweł; Kłak, Marlena; Wojtasik, Elżbieta; Górski, Andrzej

    2012-01-01

    Phage therapy (PT) is a unique method of treatment of bacterial infections using bacteriophages (phages)-viruses that specifically kill bacteria, including their antibiotic-resistant strains. Over the last decade a marked increase in interest in the therapeutic use of phages has been observed, which has resulted from a substantial rise in the prevalence of antibiotic resistance of bacteria, coupled with an inadequate number of new antibiotics. The first, and so far the only, center of PT in the European Union is the Phage Therapy Unit (PTU) established at the Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wrocław, Poland in 2005. This center continues the rich tradition of PT in Poland, which dates from the early 1920s. The main objective of this chapter is to present a detailed retrospective analysis of the results of PT of 153 patients with a wide range of infections resistant to antibiotic therapy admitted for treatment at the PTU between January 2008 and December 2010. Analysis includes the evaluation of both the efficacy and the safety of PT. In general, data suggest that PT can provide good clinical results in a significant cohort of patients with otherwise untreatable chronic bacterial infections and is essentially well tolerated. In addition, the whole complex procedure employed to obtain and characterize therapeutic phage preparations, as well as ethical aspects of PT, is discussed.

  13. Epidemiology of Campylobacter in poultry.

    NARCIS (Netherlands)

    Jacobs-Reitsma, W.

    1994-01-01

    Campylobacter , causing human infections with severe symptoms of diarrhoea, is mainly transmitted by food, especially poultry meat products.Several studies on Campylobacter colonization in breeders, laying hens, and broilers were carried out. Campylobacter isolates were serotyped, using a modificati

  14. Phenotypic resistance and the dynamics of bacterial escape from phage control

    DEFF Research Database (Denmark)

    Bull, James J.; Vegge, Christina Skovgaard; Schmerer, Matthew

    2014-01-01

    The canonical view of phage - bacterial interactions in dense, liquid cultures is that the phage will eliminate most of the sensitive cells; genetic resistance will then ascend to restore high bacterial densities. Yet there are various mechanisms by which bacteria may remain sensitive to phages...... mathematical models of these processes and suggest how different types of this 'phenotypic' resistance may be elucidated. We offer preliminary in vitro studies of a previously characterized E. coli model system and Campylobacter jejuni illustrating apparent phenotypic resistance. As phenotypic resistance may...

  15. Aerosol phage therapy efficacy in Burkholderia cepacia complex respiratory infections.

    Science.gov (United States)

    Semler, Diana D; Goudie, Amanda D; Finlay, Warren H; Dennis, Jonathan J

    2014-07-01

    Phage therapy has been suggested as a potential treatment for highly antibiotic-resistant bacteria, such as the species of the Burkholderia cepacia complex (BCC). To address this hypothesis, experimental B. cenocepacia respiratory infections were established in mice using a nebulizer and a nose-only inhalation device. Following infection, the mice were treated with one of five B. cenocepacia-specific phages delivered as either an aerosol or intraperitoneal injection. The bacterial and phage titers within the lungs were assayed 2 days after treatment, and mice that received the aerosolized phage therapy demonstrated significant decreases in bacterial loads. Differences in phage activity were observed in vivo. Mice that received phage treatment by intraperitoneal injection did not demonstrate significantly reduced bacterial loads, although phage particles were isolated from their lung tissue. Based on these data, aerosol phage therapy appears to be an effective method for treating highly antibiotic-resistant bacterial respiratory infections, including those caused by BCC bacteria.

  16. Phages of Listeria offer novel tools for diagnostics and biocontrol

    Directory of Open Access Journals (Sweden)

    Martin J Loessner

    2014-04-01

    Full Text Available Historically, bacteriophages infecting their hosts have perhaps been best known and even notorious for being a nuisance in dairy-fermentation processes. However, with the rapid progress in molecular microbiology and microbial ecology, a new dawn has risen for phages. This review will provide an overview on possible uses and applications of Listeria phages, including phage-typing, reporter phage for bacterial diagnostics, and use of phage as biocontrol agents for food safety. The use of phage-encoded enzymes such as endolysins for the detection and as antimicrobial will also be addressed. Desirable properties of candidate phages for biocontrol will be discussed. While emphasizing the enormous future potential for applications, we will also consider some of the intrinsic limitations dictated by both phage and bacterial ecology.

  17. Bacteria, phages and septicemia.

    Directory of Open Access Journals (Sweden)

    Ausra Gaidelyte

    Full Text Available The use of phages is an attractive option to battle antibiotic resistant bacteria in certain bacterial infections, but the role of phage ecology in bacterial infections is obscure. Here we surveyed the phage ecology in septicemia, the most severe type of bacterial infection. We observed that the majority of the bacterial isolates from septicemia patients spontaneously secreted phages active against other isolates of the same bacterial strain, but not to the strain causing the disease. Such phages were also detected in the initial blood cultures, indicating that phages are circulating in the blood at the onset of sepsis. The fact that most of the septicemic bacterial isolates carry functional prophages suggests an active role of phages in bacterial infections. Apparently, prophages present in sepsis-causing bacterial clones play a role in clonal selection during bacterial invasion.

  18. Phage as a modulator of immune responses: practical implications for phage therapy.

    Science.gov (United States)

    Górski, Andrzej; Międzybrodzki, Ryszard; Borysowski, Jan; Dąbrowska, Krystyna; Wierzbicki, Piotr; Ohams, Monika; Korczak-Kowalska, Grażyna; Olszowska-Zaremba, Natasza; Łusiak-Szelachowska, Marzena; Kłak, Marlena; Jończyk, Ewa; Kaniuga, Ewelina; Gołaś, Aneta; Purchla, Sylwia; Weber-Dąbrowska, Beata; Letkiewicz, Sławomir; Fortuna, Wojciech; Szufnarowski, Krzysztof; Pawełczyk, Zdzisław; Rogóż, Paweł; Kłosowska, Danuta

    2012-01-01

    Although the natural hosts for bacteriophages are bacteria, a growing body of data shows that phages can also interact with some populations of mammalian cells, especially with cells of the immune system. In general, these interactions include two main aspects. The first is the phage immunogenicity, that is, the capacity of phages to induce specific immune responses, in particular the generation of specific antibodies against phage antigens. The other aspect includes the immunomodulatory activity of phages, that is, the nonspecific effects of phages on different functions of major populations of immune cells involved in both innate and adaptive immune responses. These functions include, among others, phagocytosis and the respiratory burst of phagocytic cells, the production of cytokines, and the generation of antibodies against nonphage antigens. The aim of this chapter is to discuss the interactions between phages and cells of the immune system, along with their implications for phage therapy. These topics are presented based on the results of experimental studies and unique data on immunomodulatory effects found in patients with bacterial infections treated with phage preparations.

  19. Flying insects and Campylobacter

    DEFF Research Database (Denmark)

    Hald, Birthe; Sommer, Helle Mølgaard; Skovgård, Henrik

    to broiler houses may be counted in thousands per broiler rotation during summer periods, even a low prevalence of Campylobacter positive flies constitute a risk of introduction of Campylobacter to the chickens. M. domestica – the house fly is the most important vector fly for Campylobacter transmission...... organisms, which may collect on their bodies or survive passage through the fly gut. Campylobacter and other pathogens are then easily transferred to other surfaces, for instance peoples food – or to broiler houses where they may be swallowed by chickens or contaminate the environment. On a large material...... of several species of flies collected outside broiler houses, merely ~1% of the flies were found Campylobacter positive. However, the prevalence varied considerably with fly species, time of the year, and availability of Campylobacter sources. Influx of flies to broiler houses As the influx of flies...

  20. Phage display: concept, innovations, applications and future.

    Science.gov (United States)

    Pande, Jyoti; Szewczyk, Magdalena M; Grover, Ashok K

    2010-01-01

    Phage display is the technology that allows expression of exogenous (poly)peptides on the surface of phage particles. The concept is simple in principle: a library of phage particles expressing a wide diversity of peptides is used to select those that bind the desired target. The filamentous phage M13 is the most commonly used vector to create random peptide display libraries. Several methods including recombinant techniques have been developed to increase the diversity of the library. On the other extreme, libraries with various biases can be created for specific purposes. For instance, when the sequence of the peptide that binds the target is known, its affinity and selectivity can be increased by screening libraries created with limited mutagenesis of the peptide. Phage libraries are screened for binding to synthetic or native targets. The initial screening of library by basic biopanning has been extended to column chromatography including negative screening and competition between selected phage clones to identify high affinity ligands with greater target specificity. The rapid isolation of specific ligands by phage display is advantageous in many applications including selection of inhibitors for the active and allosteric sites of the enzymes, receptor agonists and antagonists, and G-protein binding modulatory peptides. Phage display has been used in epitope mapping and analysis of protein-protein interactions. The specific ligands isolated from phage libraries can be used in therapeutic target validation, drug design and vaccine development. Phage display can also be used in conjunction with other methods. The past innovations and those to come promise a bright future for this field.

  1. Campylobacter jejuni motility is required for infection of the flagellotropic bacteriophage F341

    DEFF Research Database (Denmark)

    Baldvinsson, Signe Berg; Sørensen, Martine Camilla Holst; Vegge, Christina Skovgaard;

    2014-01-01

    Previous studies have identified a specific modification of the capsular polysaccharide as receptor for phages that infect Campylobacter jejuni. Using acapsular kpsM mutants of C. jejuni strains NCTC11168 and NCTC12658, we found that bacteriophage F341 infects C. jejuni independently of the capsule...

  2. The effect of reducing numbers of Campylobacter in broiler intestines on human health risk

    DEFF Research Database (Denmark)

    Nauta, Maarten; Johannessen, Gro; Laureano Adame, Laura

    2016-01-01

    One option for Campylobacter control in broiler chickens is to reduce the concentration in the intestinal content of the birds prior to slaughter, for example by vaccination or phage therapy. It is however unsure how such a reduction in concentration can be translated into a reduction...

  3. Phage-Antibiotic Synergy (PAS): beta-lactam and quinolone antibiotics stimulate virulent phage growth.

    Science.gov (United States)

    Comeau, André M; Tétart, Françoise; Trojet, Sabrina N; Prère, Marie-Françoise; Krisch, H M

    2007-08-29

    Although the multiplication of bacteriophages (phages) has a substantial impact on the biosphere, comparatively little is known about how the external environment affects phage production. Here we report that sub-lethal concentrations of certain antibiotics can substantially stimulate the host bacterial cell's production of some virulent phage. For example, a low dosage of cefotaxime, a cephalosporin, increased an uropathogenic Escherichia coli strain's production of the phage PhiMFP by more than 7-fold. We name this phenomenon Phage-Antibiotic Synergy (PAS). A related effect was observed in diverse host-phage systems, including the T4-like phages, with beta-lactam and quinolone antibiotics, as well as mitomycin C. A common characteristic of these antibiotics is that they inhibit bacterial cell division and trigger the SOS system. We therefore examined the PAS effect within the context of the bacterial SOS and filamentation responses. We found that the PAS effect appears SOS-independent and is primarily a consequence of cellular filamentation; it is mimicked by cells that constitutively filament. The fact that completely unrelated phages manifest this phenomenon suggests that it confers an important and general advantage to the phages.

  4. Phage-Antibiotic Synergy (PAS: beta-lactam and quinolone antibiotics stimulate virulent phage growth.

    Directory of Open Access Journals (Sweden)

    André M Comeau

    Full Text Available Although the multiplication of bacteriophages (phages has a substantial impact on the biosphere, comparatively little is known about how the external environment affects phage production. Here we report that sub-lethal concentrations of certain antibiotics can substantially stimulate the host bacterial cell's production of some virulent phage. For example, a low dosage of cefotaxime, a cephalosporin, increased an uropathogenic Escherichia coli strain's production of the phage PhiMFP by more than 7-fold. We name this phenomenon Phage-Antibiotic Synergy (PAS. A related effect was observed in diverse host-phage systems, including the T4-like phages, with beta-lactam and quinolone antibiotics, as well as mitomycin C. A common characteristic of these antibiotics is that they inhibit bacterial cell division and trigger the SOS system. We therefore examined the PAS effect within the context of the bacterial SOS and filamentation responses. We found that the PAS effect appears SOS-independent and is primarily a consequence of cellular filamentation; it is mimicked by cells that constitutively filament. The fact that completely unrelated phages manifest this phenomenon suggests that it confers an important and general advantage to the phages.

  5. Phage typing of Staphylococcus saprophyticus.

    Science.gov (United States)

    Torres Pereira, A.; Melo Cristino, J. A.

    1991-01-01

    This study included 502 staphylococcus strains; Staphylococcus saprophyticus (297 strains) S. cohnii (47), S. xylosus (10), S. epidermidis (67) and S. aureus (81). Mitomycin C induction was performed on 100 isolates of S. saprophyticus and all induced strains were reacted with each other. Twenty-six strains proved to be lysogenic. Phages were propagated and titrated. With 12 of the phages there were three frequent associations, named lytic groups A, B and C, which included 75% of all typable strains. Typability of the system was 45% and reproducibility was between 94.2% and 100%. Phages did not lyse S. aureus and S. epidermidis strains, but they lysed S. saprophyticus and only rare strains of other novobiocin resistant species. Effective S. saprophyticus typing serves ecological purposes and tracing the origin of urinary strains from the skin or mucous membranes. Phage typing in association with plasmid profiling previously described, are anticipated as complementary methods with strong discriminatory power for differentiating among S. saprophyticus strains. PMID:1752305

  6. Major structural differences and novel potential virulence mechanisms from the genomes of multiple campylobacter species.

    Directory of Open Access Journals (Sweden)

    Derrick E Fouts

    2005-01-01

    Full Text Available Sequencing and comparative genome analysis of four strains of Campylobacter including C. lari RM2100, C. upsaliensis RM3195, and C. coli RM2228 has revealed major structural differences that are associated with the insertion of phage- and plasmid-like genomic islands, as well as major variations in the lipooligosaccharide complex. Poly G tracts are longer, are greater in number, and show greater variability in C. upsaliensis than in the other species. Many genes involved in host colonization, including racR/S, cadF, cdt, ciaB, and flagellin genes, are conserved across the species, but variations that appear to be species specific are evident for a lipooligosaccharide locus, a capsular (extracellular polysaccharide locus, and a novel Campylobacter putative licABCD virulence locus. The strains also vary in their metabolic profiles, as well as their resistance profiles to a range of antibiotics. It is evident that the newly identified hypothetical and conserved hypothetical proteins, as well as uncharacterized two-component regulatory systems and membrane proteins, may hold additional significant information on the major differences in virulence among the species, as well as the specificity of the strains for particular hosts.

  7. Fluoroquinolone resistance in Campylobacter

    Science.gov (United States)

    Fluoroquinolone-resistant Campylobacter jejuni and C. coli are common in animals because of the use of fluoroquinolones as therapeutic agents in animal husbandry, particularly in chickens and other poultry. Campylobacter is a commensal in poultry, and therefore, poultry and poultry products are the...

  8. Campylobacter virulence and survival factors.

    Science.gov (United States)

    Bolton, Declan J

    2015-06-01

    Despite over 30 years of research, campylobacteriosis is the most prevalent foodborne bacterial infection in many countries including in the European Union and the United States of America. However, relatively little is known about the virulence factors in Campylobacter or how an apparently fragile organism can survive in the food chain, often with enhanced pathogenicity. This review collates information on the virulence and survival determinants including motility, chemotaxis, adhesion, invasion, multidrug resistance, bile resistance and stress response factors. It discusses their function in transition through the food processing environment and human infection. In doing so it provides a fundamental understanding of Campylobacter, critical for improved diagnosis, surveillance and control. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Flying insects and Campylobacter

    DEFF Research Database (Denmark)

    Hald, Birthe; Sommer, Helle Mølgaard; Skovgård, Henrik

    Campylobacter in flies Flies of the Muscidae family forage on all kind of faeces – various fly species have different preferences. M domestica prefer pigs, horses and cattle faeces, animals which are all known to frequently excrete Campylobacter. As a result, the insects pick up pathogenic micro...... organisms, which may collect on their bodies or survive passage through the fly gut. Campylobacter and other pathogens are then easily transferred to other surfaces, for instance peoples food – or to broiler houses where they may be swallowed by chickens or contaminate the environment. On a large material...... of several species of flies collected outside broiler houses, merely ~1% of the flies were found Campylobacter positive. However, the prevalence varied considerably with fly species, time of the year, and availability of Campylobacter sources. Influx of flies to broiler houses As the influx of flies...

  10. Phage lytic enzymes: a history

    Institute of Scientific and Technical Information of China (English)

    David; Trudil

    2015-01-01

    There are many recent studies regarding the efficacy of bacteriophage-related lytic enzymes: the enzymes of ‘bacteria-eaters’ or viruses that infect bacteria. By degrading the cell wall of the targeted bacteria, these lytic enzymes have been shown to efficiently lyse Gram-positive bacteria without affecting normal flora and non-related bacteria. Recent studies have suggested approaches for lysing Gram-negative bacteria as well(Briersa Y, et al., 2014). These enzymes include: phage-lysozyme, endolysin, lysozyme, lysin, phage lysin, phage lytic enzymes, phageassociated enzymes, enzybiotics, muralysin, muramidase, virolysin and designations such as Ply, PAE and others. Bacteriophages are viruses that kill bacteria, do not contribute to antimicrobial resistance, are easy to develop, inexpensive to manufacture and safe for humans, animals and the environment. The current focus on lytic enzymes has been on their use as anti-infectives in humans and more recently in agricultural research models. The initial translational application of lytic enzymes, however, was not associated with treating or preventing a specifi c disease but rather as an extraction method to be incorporated in a rapid bacterial detection assay(Bernstein D, 1997).The current review traces the translational history of phage lytic enzymes–from their initial discovery in 1986 for the rapid detection of group A streptococcus in clinical specimens to evolving applications in the detection and prevention of disease in humans and in agriculture.

  11. Control of campylobacter in poultry industry from farm to poultry processing unit: A review.

    Science.gov (United States)

    Umaraw, Pramila; Prajapati, A; Verma, Akhilesh K; Pathak, V; Singh, V P

    2017-03-04

    Campylobacter is an emerging zoonotic bacterial threat in the poultry industry. Most of the human cases of campylobacteriosis recorded have revealed their poultry origins. Various control measures have been employed both at the farm and processing levels to combat with it. The antibiotic treatment, phage therapy, competitive exclusion, and vaccination have been adapted at the farm level to reduce colonization of Campylobacter in poultry gut. While prevention of intestinal spillage, scheduled slaughter, logistic slaughter, chemical decontamination of carcasses are recommended to reduce contamination during processing. The postharvest interventions such as heat treatment, freezing, irradiation of contaminated carcass can effectively reduce Campylobacter contamination. Thus, integrated approaches are required to tackle infection of Campylobacter in humans.

  12. Effect of Noradrenaline on the Virulence Properties of Campylobacter Species

    Directory of Open Access Journals (Sweden)

    Sree V. Aroori

    2014-01-01

    Full Text Available Campylobacter species cause a spectrum of illnesses in humans. The type of illness and the outcome is dependent on the virulence of the infecting pathogen strain and host immune status. Acute stress can seriously compromise host immunity and increase susceptibility to infection. Noradrenaline (NA is a stress hormone. Several studies have shown that it stimulated growth and increased the pathogenicity of organisms including E. coli and Campylobacter jejuni. However, the effect of NA on other Campylobacter species is unknown. We have examined the effect of NA on growth rate, motility, invasion of T84 epithelial cells, and colonisation of chickens by diverse Campylobacter species. Campylobacter cultures grown with NA had reduced lag phases, increased growth rates, and higher final optical densities than controls. The motility of Campylobacter was also significantly increased in the presence of noradrenaline. Some of the Campylobacter strains tested also showed increased invasion of T84 epithelial cells, greater breakdown of tight junctions, and an enhanced potential to colonise chickens. Our results show that noradrenaline-induced enhancement of virulence of Campylobacter can influence the outcome of infection.

  13. Phage therapy: present and future

    Science.gov (United States)

    Kolesnikova, S. G.; Tulyakova, E. N.; Moiseeva, I. Y.

    2017-01-01

    In recent years, bacteriophages are known to have become an effective alternative to antibiotic drugs. The article describes the current and potential applications of bacteriophages and phage endolysins. Also of interest is the devastating effect of phages on biofilms. The development of phage resistance is touched upon as well. Furthermore, the authors discuss the issue of laying down the rules of rational phage therapy.

  14. Estimating richness from phage metagenomes

    Science.gov (United States)

    Bacteriophages are important drivers of ecosystem functions, yet little is known about the vast majority of phages. Phage metagenomics, or the study of the collective genome of an assemblage of phages, enables the investigation of broad ecological questions in phage communities. One ecological cha...

  15. Phage choice, isolation, and preparation for phage therapy.

    Science.gov (United States)

    Gill, Jason J; Hyman, Paul

    2010-01-01

    Phage therapy is the use of bacteriophages--viruses that use bacteria as their host cells--as biocontrol agents of bacteria. Currently, phage therapy is garnering renewed interest as bacterial resistance to antibiotics becomes widespread. Historically, phage therapy was largely abandoned in the West in the 1940s due to the advent of chemical antibiotics, and the unreliability of phage-based treatments when compared to antibiotics. The choice of phage strain and the methods of phage preparation are now thought to have been critical to the success or failure of phage therapy trials. Insufficiently virulent phages, especially against actual target bacteria, allow bacteria to survive treatment while poorly prepared phage stocks, even if of sufficiently virulent phages, lack the numbers of viable phages required for adequate treatment. In this review we discuss the factors that determine the methods of isolation, analysis, and identification of phage species for phage therapy. We go on to discuss the various methods available for purifying phages as well as considerations of the degree of purification which is sufficient for various applications. Lastly, we review the current practices used to prepare commercial phage therapy products.

  16. Media for Campylobacter jejuni and other campylobacters

    NARCIS (Netherlands)

    Mossel, D.A.A.

    1985-01-01

    Despite their recent elaboration and the many variations in antibiotic combinations designed to attain selectivity, highly selective liquid and solid culture media for Campylobacter jejuni have proved satisfactory provided they are incubated at about 42°C and in a microaerophilic atmosphere such as

  17. Media for Campylobacter jejuni and other campylobacters

    NARCIS (Netherlands)

    Mossel, D.A.A.

    1985-01-01

    Despite their recent elaboration and the many variations in antibiotic combinations designed to attain selectivity, highly selective liquid and solid culture media for Campylobacter jejuni have proved satisfactory provided they are incubated at about 42°C and in a microaerophilic atmosphere such as

  18. Supersize me: Cronobacter sakazakii phage GAP32

    Energy Technology Data Exchange (ETDEWEB)

    Abbasifar, Reza; Griffiths, Mansel W. [Canadian Research Institute for Food Safety, University of Guelph, Guelph, ON, Canada N1G 2W1 (Canada); Sabour, Parviz M. [Agriculture and Agri-Food Canada, Guelph Food Research Centre, Guelph, ON, Canada N1G 5C9 (Canada); Ackermann, Hans-Wolfgang [Department of Microbiology-Infectiology and Immunology, Faculty of Medicine, Université Laval, Quebec, QC (Canada); Vandersteegen, Katrien; Lavigne, Rob [Laboratory of Gene Technology, Katholieke Universiteit Leuven, Leuven (Belgium); Noben, Jean-Paul [Biomedical Research Institute and Transnational University Limburg, School of Life Sciences, Hasselt University, Diepenbeek (Belgium); Alanis Villa, Argentina; Abbasifar, Arash [Canadian Research Institute for Food Safety, University of Guelph, Guelph, ON, Canada N1G 2W1 (Canada); Nash, John H.E. [Public Health Agency of Canada, Laboratory for Foodborne Zoonoses, Guelph, ON, Canada N1G 3W4 (Canada); Kropinski, Andrew M., E-mail: akropins@uoguelph.ca [Public Health Agency of Canada, Laboratory for Foodborne Zoonoses, Guelph, ON, Canada N1G 3W4 (Canada); Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada N1G 2W1 (Canada)

    2014-07-15

    Cronobacter sakazakii is a Gram-negative pathogen found in milk-based formulae that causes infant meningitis. Bacteriophages have been proposed to control bacterial pathogens; however, comprehensive knowledge about a phage is required to ensure its safety before clinical application. We have characterized C. sakazakii phage vB{sub C}saM{sub G}AP32 (GAP32), which possesses the second largest sequenced phage genome (358,663 bp). A total of 571 genes including 545 protein coding sequences and 26 tRNAs were identified, thus more genes than in the smallest bacterium, Mycoplasma genitalium G37. BLASTP and HHpred searches, together with proteomic analyses reveal that only 23.9% of the putative proteins have defined functions. Some of the unique features of this phage include: a chromosome condensation protein, two copies of the large subunit terminase, a predicted signal-arrest-release lysin; and an RpoD-like protein, which is possibly involved in the switch from immediate early to delayed early transcription. Its closest relatives are all extremely large myoviruses, namely coliphage PBECO4 and Klebsiella phage vB{sub K}leM-RaK2, with whom it shares approximately 44% homologous proteins. Since the homologs are not evenly distributed, we propose that these three phages belong to a new subfamily. - Highlights: • Cronobacter sakazakii phage vB{sub C}saM{sub G}AP32 has a genome of 358,663 bp. • It encodes 545 proteins which is more than Mycoplasma genitalium G37. • It is a member of the Myoviridae. • It is peripherally related to coliphage PBECO4 and Klebsiella phage vB{sub K}leM-RaK2. • GAP32 encodes a chromosome condensation protein.

  19. Convergent evolution of pathogenicity islands in helper cos phage interference

    Science.gov (United States)

    Manning, Keith A.; Dokland, Terje; Marina, Alberto

    2016-01-01

    Staphylococcus aureus pathogenicity islands (SaPIs) are phage satellites that exploit the life cycle of their helper phages for their own benefit. Most SaPIs are packaged by their helper phages using a headful (pac) packaging mechanism. These SaPIs interfere with pac phage reproduction through a variety of strategies, including the redirection of phage capsid assembly to form small capsids, a process that depends on the expression of the SaPI-encoded cpmA and cpmB genes. Another SaPI subfamily is induced and packaged by cos-type phages, and although these cos SaPIs also block the life cycle of their inducing phages, the basis for this mechanism of interference remains to be deciphered. Here we have identified and characterized one mechanism by which the SaPIs interfere with cos phage reproduction. This mechanism depends on a SaPI-encoded gene, ccm, which encodes a protein involved in the production of small isometric capsids, compared with the prolate helper phage capsids. As the Ccm and CpmAB proteins are completely unrelated in sequence, this strategy represents a fascinating example of convergent evolution. Moreover, this result also indicates that the production of SaPI-sized particles is a widespread strategy of phage interference conserved during SaPI evolution. This article is part of the themed issue ‘The new bacteriology’. PMID:27672154

  20. Convergent evolution of pathogenicity islands in helper cos phage interference.

    Science.gov (United States)

    Carpena, Nuria; Manning, Keith A; Dokland, Terje; Marina, Alberto; Penadés, José R

    2016-11-05

    Staphylococcus aureus pathogenicity islands (SaPIs) are phage satellites that exploit the life cycle of their helper phages for their own benefit. Most SaPIs are packaged by their helper phages using a headful (pac) packaging mechanism. These SaPIs interfere with pac phage reproduction through a variety of strategies, including the redirection of phage capsid assembly to form small capsids, a process that depends on the expression of the SaPI-encoded cpmA and cpmB genes. Another SaPI subfamily is induced and packaged by cos-type phages, and although these cos SaPIs also block the life cycle of their inducing phages, the basis for this mechanism of interference remains to be deciphered. Here we have identified and characterized one mechanism by which the SaPIs interfere with cos phage reproduction. This mechanism depends on a SaPI-encoded gene, ccm, which encodes a protein involved in the production of small isometric capsids, compared with the prolate helper phage capsids. As the Ccm and CpmAB proteins are completely unrelated in sequence, this strategy represents a fascinating example of convergent evolution. Moreover, this result also indicates that the production of SaPI-sized particles is a widespread strategy of phage interference conserved during SaPI evolution.This article is part of the themed issue 'The new bacteriology'.

  1. Bacteriophage receptor binding protein based assays for the simultaneous detection of Campylobacter jejuni and Campylobacter coli.

    Science.gov (United States)

    Javed, Muhammad A; Poshtiban, Somayyeh; Arutyunov, Denis; Evoy, Stephane; Szymanski, Christine M

    2013-01-01

    Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of foodborne gastroenteritis which is occasionally followed by a debilitating neuropathy known as Guillain-Barré syndrome. Rapid and specific detection of these pathogens is very important for effective control and quick treatment of infection. Most of the diagnostics available for these organisms are time consuming and require technical expertise with expensive instruments and reagents to perform. Bacteriophages bind to their host specifically through their receptor binding proteins (RBPs), which can be exploited for pathogen detection. We recently sequenced the genome of C. jejuni phage NCTC12673 and identified its putative host receptor binding protein, Gp047. In the current study, we localized the receptor binding domain to the C-terminal quarter of Gp047. CC-Gp047 could be produced recombinantly and was capable of agglutinating both C. jejuni and C. coli cells unlike the host range of the parent phage which is limited to a subset of C. jejuni isolates. The agglutination procedure could be performed within minutes on a glass slide at room temperature and was not hindered by the presence of buffers or nutrient media. This agglutination assay showed 100% specificity and the sensitivity was 95% for C. jejuni (n = 40) and 90% for C. coli (n = 19). CC-Gp047 was also expressed as a fusion with enhanced green fluorescent protein (EGFP). Chimeric EGFP_CC-Gp047 was able to specifically label C. jejuni and C. coli cells in mixed cultures allowing for the detection of these pathogens by fluorescent microscopy. This study describes a simple and rapid method for the detection of C. jejuni and C. coli using engineered phage RBPs and offers a promising new diagnostics platform for healthcare and surveillance laboratories.

  2. Bacteriophage receptor binding protein based assays for the simultaneous detection of Campylobacter jejuni and Campylobacter coli.

    Directory of Open Access Journals (Sweden)

    Muhammad A Javed

    Full Text Available Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of foodborne gastroenteritis which is occasionally followed by a debilitating neuropathy known as Guillain-Barré syndrome. Rapid and specific detection of these pathogens is very important for effective control and quick treatment of infection. Most of the diagnostics available for these organisms are time consuming and require technical expertise with expensive instruments and reagents to perform. Bacteriophages bind to their host specifically through their receptor binding proteins (RBPs, which can be exploited for pathogen detection. We recently sequenced the genome of C. jejuni phage NCTC12673 and identified its putative host receptor binding protein, Gp047. In the current study, we localized the receptor binding domain to the C-terminal quarter of Gp047. CC-Gp047 could be produced recombinantly and was capable of agglutinating both C. jejuni and C. coli cells unlike the host range of the parent phage which is limited to a subset of C. jejuni isolates. The agglutination procedure could be performed within minutes on a glass slide at room temperature and was not hindered by the presence of buffers or nutrient media. This agglutination assay showed 100% specificity and the sensitivity was 95% for C. jejuni (n = 40 and 90% for C. coli (n = 19. CC-Gp047 was also expressed as a fusion with enhanced green fluorescent protein (EGFP. Chimeric EGFP_CC-Gp047 was able to specifically label C. jejuni and C. coli cells in mixed cultures allowing for the detection of these pathogens by fluorescent microscopy. This study describes a simple and rapid method for the detection of C. jejuni and C. coli using engineered phage RBPs and offers a promising new diagnostics platform for healthcare and surveillance laboratories.

  3. Precisely modulated pathogenicity island interference with late phage gene transcription.

    Science.gov (United States)

    Ram, Geeta; Chen, John; Ross, Hope F; Novick, Richard P

    2014-10-07

    Having gone to great evolutionary lengths to develop resistance to bacteriophages, bacteria have come up with resistance mechanisms directed at every aspect of the bacteriophage life cycle. Most genes involved in phage resistance are carried by plasmids and other mobile genetic elements, including bacteriophages and their relatives. A very special case of phage resistance is exhibited by the highly mobile phage satellites, staphylococcal pathogenicity islands (SaPIs), which carry and disseminate superantigen and other virulence genes. Unlike the usual phage-resistance mechanisms, the SaPI-encoded interference mechanisms are carefully crafted to ensure that a phage-infected, SaPI-containing cell will lyse, releasing the requisite crop of SaPI particles as well as a greatly diminished crop of phage particles. Previously described SaPI interference genes target phage functions that are not required for SaPI particle production and release. Here we describe a SaPI-mediated interference system that affects expression of late phage gene transcription and consequently is required for SaPI and phage. Although when cloned separately, a single SaPI gene totally blocks phage production, its activity in situ is modulated accurately by a second gene, achieving the required level of interference. The advantage for the host bacteria is that the SaPIs curb excessive phage growth while enhancing their gene transfer activity. This activity is in contrast to that of the clustered regularly interspaced short palindromic repeats (CRISPRs), which totally block phage growth at the cost of phage-mediated gene transfer. In staphylococci the SaPI strategy seems to have prevailed during evolution: The great majority of Staphylococcus aureus strains carry one or more SaPIs, whereas CRISPRs are extremely rare.

  4. Campylobacter jejuni motility is required for infection of the flagellotropic bacteriophage F341.

    Science.gov (United States)

    Baldvinsson, Signe Berg; Sørensen, Martine C Holst; Vegge, Christina S; Clokie, Martha R J; Brøndsted, Lone

    2014-11-01

    Previous studies have identified a specific modification of the capsular polysaccharide as receptor for phages that infect Campylobacter jejuni. Using acapsular kpsM mutants of C. jejuni strains NCTC11168 and NCTC12658, we found that bacteriophage F341 infects C. jejuni independently of the capsule. In contrast, phage F341 does not infect C. jejuni NCTC11168 mutants that either lack the flagellar filaments (ΔflaAB) or that have paralyzed, i.e., nonrotating, flagella (ΔmotA and ΔflgP). Complementing flgP confirmed that phage F341 requires rotating flagella for successful infection. Furthermore, adsorption assays demonstrated that phage F341 does not adsorb to these nonmotile C. jejuni NCTC11168 mutants. Taken together, we propose that phage F341 uses the flagellum as a receptor. Phage-host interactions were investigated using fluorescence confocal and transmission electron microscopy. These data demonstrate that F341 binds to the flagellum by perpendicular attachment with visible phage tail fibers interacting directly with the flagellum. Our data are consistent with the movement of the C. jejuni flagellum being required for F341 to travel along the filament to reach the basal body of the bacterium. The initial binding to the flagellum may cause a conformational change of the phage tail that enables DNA injection after binding to a secondary receptor.

  5. Campylobacter jejuni Motility Is Required for Infection of the Flagellotropic Bacteriophage F341

    Science.gov (United States)

    Baldvinsson, Signe Berg; Sørensen, Martine C. Holst; Vegge, Christina S.; Clokie, Martha R. J.

    2014-01-01

    Previous studies have identified a specific modification of the capsular polysaccharide as receptor for phages that infect Campylobacter jejuni. Using acapsular kpsM mutants of C. jejuni strains NCTC11168 and NCTC12658, we found that bacteriophage F341 infects C. jejuni independently of the capsule. In contrast, phage F341 does not infect C. jejuni NCTC11168 mutants that either lack the flagellar filaments (ΔflaAB) or that have paralyzed, i.e., nonrotating, flagella (ΔmotA and ΔflgP). Complementing flgP confirmed that phage F341 requires rotating flagella for successful infection. Furthermore, adsorption assays demonstrated that phage F341 does not adsorb to these nonmotile C. jejuni NCTC11168 mutants. Taken together, we propose that phage F341 uses the flagellum as a receptor. Phage-host interactions were investigated using fluorescence confocal and transmission electron microscopy. These data demonstrate that F341 binds to the flagellum by perpendicular attachment with visible phage tail fibers interacting directly with the flagellum. Our data are consistent with the movement of the C. jejuni flagellum being required for F341 to travel along the filament to reach the basal body of the bacterium. The initial binding to the flagellum may cause a conformational change of the phage tail that enables DNA injection after binding to a secondary receptor. PMID:25261508

  6. Campylobacter spp. as a foodborne pathogen: a review

    Directory of Open Access Journals (Sweden)

    Joana eSilva

    2011-09-01

    Full Text Available Campylobacter is well recognized as the leading cause of bacterial foodborne diarrheal disease worldwide, causing mild to severe symptoms including serious infections of the extremities and permanent neurological symptoms. The organism is a cytochrome oxidase positive, microaerophilic, curved Gram-negative rod exhibiting corkscrew motility and is carried in the intestine of many wild and domestic animals, particularly avian species including poultry, where the intestine is colonized resulting in healthy animals as carriers. This review aims to elucidate and discuss the i genus Campylobacter, growth and survival characteristics; ii detection, isolation and confirmation of Campylobacter; iii campylobacteriosis and presence of virulence factors and iv colonization of poultry and control strategies.

  7. Recovery of Campylobacter jejuni and Campylobacter coli from inoculated foods by selective enrichment.

    Science.gov (United States)

    Doyle, M P; Roman, D J

    1982-06-01

    A direct enrichment procedure was developed to selectively recover small numbers of Campylobacter jejuni, C. coli, and nalidixic acid-resistant thermophilic Campylobacter from foods. The procedure includes an enrichment medium composed of brucella broth, 7% lysed horse blood, 0.3% sodium succinate, 0.01% cysteine hydrochloride, vancomycin (15 micrograms/ml), trimethoprim (5 micrograms/ml), polymyxin B (20 IU/ml), and cycloheximide (50 micrograms/ml) that is inoculated with 10 or 25 g of food and incubated with agitation under microaerophilic conditions at 42 degrees C for 16 to 18 h. After incubation, the medium is plated directly onto Campy-BAP agar plates (M. J. Blaser et al., Ann. Intern. Med. 91:179-185, 1979), and resulting colonies that resemble Campylobacter are identified by conventional tests. The foods evaluated included raw milk, hamburger, and chicken skin which had aerobic plate counts of 10(5) to 10(9) bacteria/g. The procedure was effective in recovering as few as 0.1 cell of Campylobacter per g of food. Of the 50 isolates of Campylobacter evaluated, all were recovered from raw milk and hamburger at a level of 1 to 4 cells/g, and 41 and 40 isolaes were recovered from the hamburger and milk, respectively, at 0.1 to 0.4 cell/g. The enrichment was least effective for recovering campylobacters from chicken skin, as 7 and 26 of 50 isolates were not recovered at 1 to 4 and 0.1 to 0.4 cell/g, respectively. This new procedure is more rapid, direct, and effective than other enrichment or direct plating procedures for recovering small numbers of campylobacters from foods.

  8. Participation of some campylobacter species in the etiology of enterocolitis

    Directory of Open Access Journals (Sweden)

    Otašević Marica M.

    2004-01-01

    Full Text Available Background. In recent decades, medical community has increasingly been calling attention to the importance of Campylobacter as an disease-causing agent in humans. Nowdays, Campylobacter jejuni (C. jejuni is known as the most frequent bacterial cause of diarrhea worldwide. Epidemiological differences of the infections caused by Campylobacter, present in the developed and the developing countries, are attributed to the differences of the types of virulence. Due to the specificity, and the demanding features of Campylobacter, as well as poorly equipped microbiological laboratories, campylobacteriosis is insufficiently studied in our country. This investigation aimed to determine the participation of some Campylobacter species in the etiology of diarrheal diseases in our population. Methods. The four-years continuous monitoring of Campylobacter presence was performed in the faeces of 12 605 patients with enterocolitis. The control group included 5 774 examinees of healthy children and youth. Faeces samples were cultivated on Skirrow's selective medium, and further incubated according to effective methodology for Campylobacter. Identification of strains was based on morphological, cultural and physiologic features of strains (oxidase test, catalase test, susceptibility to nalidixic acid, and hypurate hydrolysis. As a statistical method, for data processing, c2 test and Fisher’s exact test were used. Results. Campylobacter was proven in 3.86% of enterocolitis patients, and in 0.71% of healthy population. Out of 518 Campylobacter isolates, 86.48% belonged to enterocolitis outpatients, and 13,51% to inpatients. Predominant symptoms of the disease were diarrhea (81.83%, increased temperature (34.71%, vomiting (19.77%, and stomach pain (15.17%. The diseased were predominantly infants in the first year of life. Out of 300 Campylobacter isolates, 75% were identified as Campylobacer jejuni, 23% as Campylobacter coli (C. coli, and 2% as Campylobacter lari

  9. Campylobacter fetus infections in humans : exposure and disease

    NARCIS (Netherlands)

    Wagenaar, Jaap A; van Bergen, Marcel A P; Blaser, Martin J; Tauxe, Robert V; Newell, Diane G; van Putten, Jos P M

    2014-01-01

    Campylobacter fetus can cause intestinal illness and, occasionally, severe systemic infections. Infections mainly affect persons at higher risk, including elderly and immunocompromised individuals and those with occupational exposure to infected animals. Outbreaks are infrequent but have provided in

  10. Quality and safety requirements for sustainable phage therapy products.

    Science.gov (United States)

    Pirnay, Jean-Paul; Blasdel, Bob G; Bretaudeau, Laurent; Buckling, Angus; Chanishvili, Nina; Clark, Jason R; Corte-Real, Sofia; Debarbieux, Laurent; Dublanchet, Alain; De Vos, Daniel; Gabard, Jérôme; Garcia, Miguel; Goderdzishvili, Marina; Górski, Andrzej; Hardcastle, John; Huys, Isabelle; Kutter, Elizabeth; Lavigne, Rob; Merabishvili, Maia; Olchawa, Ewa; Parikka, Kaarle J; Patey, Olivier; Pouilot, Flavie; Resch, Gregory; Rohde, Christine; Scheres, Jacques; Skurnik, Mikael; Vaneechoutte, Mario; Van Parys, Luc; Verbeken, Gilbert; Zizi, Martin; Van den Eede, Guy

    2015-07-01

    The worldwide antibiotic crisis has led to a renewed interest in phage therapy. Since time immemorial phages control bacterial populations on Earth. Potent lytic phages against bacterial pathogens can be isolated from the environment or selected from a collection in a matter of days. In addition, phages have the capacity to rapidly overcome bacterial resistances, which will inevitably emerge. To maximally exploit these advantage phages have over conventional drugs such as antibiotics, it is important that sustainable phage products are not submitted to the conventional long medicinal product development and licensing pathway. There is a need for an adapted framework, including realistic production and quality and safety requirements, that allows a timely supplying of phage therapy products for 'personalized therapy' or for public health or medical emergencies. This paper enumerates all phage therapy product related quality and safety risks known to the authors, as well as the tests that can be performed to minimize these risks, only to the extent needed to protect the patients and to allow and advance responsible phage therapy and research.

  11. Microplate-test for the rapid determination of bacteriophage-susceptibility of Campylobacter isolates-development and validation.

    Directory of Open Access Journals (Sweden)

    Samuel Fischer

    Full Text Available A simple susceptibility test using 800 isolates of one Campylobacter strain with different degrees of susceptibility and four bacteriophages of the British phage typing scheme was developed and examined for its suitability. The test presented is economically cheaper and less time consuming than the conventional agar overlay plate assay and therefore enables the monitoring of changes in the susceptibility pattern during phage therapy under practical field conditions. The main objective of this study was to compare the simplified test with the conventional agar overlay plate assay. The conventional test describes for a population of Campylobacter: i. the rate of resistant isolates (0 plaques and ii. the degree of susceptibility, also called relative efficiency of plating (EOP, for the remaining isolates. The simplified test divides the isolates into four susceptibility ranks, which are easily distinguishable to the naked eye. Ten Campylobacter isolates out of each rank were subjected to the conventional method for validation of the simplified test. Each resistance rank contained isolates showing certain degrees of susceptibility, reflecting decreasing susceptibility by an increase of the rank. Thus, the simplified test correlated well with the conventional method. Nevertheless, it can be suggested for a clear cut to summarise the first thee ranks as "high susceptible" and to mark out the fourth rank as reduced susceptible. Further test improvements will enable the monitoring of the degree of susceptibility and potentially also of resistance during phage therapy in the field. To ensure a long-lasting successful use of phage therapy, further studies on both the loss of susceptibility and the development of resistance of Campylobacter against phages combined with their impact on phage therapy will be necessary.

  12. Genome characteristics of a novel phage from Bacillus thuringiensis showing high similarity with phage from Bacillus cereus.

    Directory of Open Access Journals (Sweden)

    Yihui Yuan

    Full Text Available Bacillus thuringiensis is an important entomopathogenic bacterium belongs to the Bacillus cereus group, which also includes B. anthracis and B. cereus. Several genomes of phages originating from this group had been sequenced, but no genome of Siphoviridae phage from B. thuringiensis has been reported. We recently sequenced and analyzed the genome of a novel phage, BtCS33, from a B. thuringiensis strain, subsp. kurstaki CS33, and compared the gneome of this phage to other phages of the B. cereus group. BtCS33 was the first Siphoviridae phage among the sequenced B. thuringiensis phages. It produced small, turbid plaques on bacterial plates and had a narrow host range. BtCS33 possessed a linear, double-stranded DNA genome of 41,992 bp with 57 putative open reading frames (ORFs. It had a typical genome structure consisting of three modules: the "late" region, the "lysogeny-lysis" region and the "early" region. BtCS33 exhibited high similarity with several phages, B. cereus phage Wβ and some variants of Wβ, in genome organization and the amino acid sequences of structural proteins. There were two ORFs, ORF22 and ORF35, in the genome of BtCS33 that were also found in the genomes of B. cereus phage Wβ and may be involved in regulating sporulation of the host cell. Based on these observations and analysis of phylogenetic trees, we deduced that B. thuringiensis phage BtCS33 and B. cereus phage Wβ may have a common distant ancestor.

  13. Detection of Campylobacter species using monoclonal antibodies

    Science.gov (United States)

    Young, Colin R.; Lee, Alice; Stanker, Larry H.

    1999-01-01

    A panel of species specific monoclonal antibodies were raised to Campylobacter coli, Campylobacter jejuni and Campylobacter lari. The isotypes, and cross-reactivity profiles of each monoclonal antibody against an extensive panel of micro- organisms, were determined.

  14. Campylobacter Risk Analysis

    DEFF Research Database (Denmark)

    Nauta, Maarten

    In several countries quantitative microbiological risk assessments (QMRAs) have been performed for Campylobacter in chicken meat. The models constructed for this purpose provide a good example of the development of QMRA in general and illustrate the diversity of available methods. Despite...... the differences between the models, the most prominent conclusions of the QMRAs are similar. These conclusions for example relate to the large risk of highly contaminated meat products and the insignificance of contamination from Campylobacter positive flocks to negative flocks during slaughter and processing...

  15. Computational models of populations of bacteria and lytic phage.

    Science.gov (United States)

    Krysiak-Baltyn, Konrad; Martin, Gregory J O; Stickland, Anthony D; Scales, Peter J; Gras, Sally L

    2016-11-01

    The use of phages to control and reduce numbers of unwanted bacteria can be traced back to the early 1900s, when phages were explored as a tool to treat infections before the wide scale use of antibiotics. Recently, phage therapy has received renewed interest as a method to treat multiresistant bacteria. Phages are also widely used in the food industry to prevent the growth of certain bacteria in foods, and are currently being explored as a tool for use in bioremediation and wastewater treatment. Despite the large body of biological research on phages, relatively little attention has been given to computational modeling of the population dynamics of phage and bacterial interactions. The earliest model was described by Campbell in the 1960s. Subsequent modifications to this model include partial or complete resistance, multiple phage binding sites, and spatial heterogeneity. This review provides a general introduction to modeling of the population dynamics of bacteria and phage. The review introduces the basic model and relevant concepts and evaluates more complex variations of the basic model published to date, including a model of disease epidemics caused by infectious bacteria. Finally, the shortcomings and potential ways to improve the models are discussed.

  16. Nieuw vaccin tegen campylobacter

    NARCIS (Netherlands)

    Wagenaar, J.A.

    2008-01-01

    Het vaccin dat de kip moet beschermen tegen de bacterie Campylobacter werkt in het laboratorium. Dat wil bacterioloog Jaap Wagenaar wel kwijt. Wanneer het er komt en zelfs of het er komt, daarover laat Wagenaar zich niet uit. "Het is een hele klus om het immuunsysteem van kippen effectief op te

  17. Nieuw vaccin tegen campylobacter

    NARCIS (Netherlands)

    Wagenaar, J.A.

    2008-01-01

    Het vaccin dat de kip moet beschermen tegen de bacterie Campylobacter werkt in het laboratorium. Dat wil bacterioloog Jaap Wagenaar wel kwijt. Wanneer het er komt en zelfs of het er komt, daarover laat Wagenaar zich niet uit. "Het is een hele klus om het immuunsysteem van kippen effectief op te late

  18. Campylobacter-Acanthamoeba interactions.

    Science.gov (United States)

    Vieira, Ana; Seddon, Alan M; Karlyshev, Andrey V

    2015-05-01

    Campylobacter jejuni is a foodborne pathogen recognized as the major cause of human bacterial enteritis. Undercooked poultry products and contaminated water are considered as the most important sources of infection. Some studies suggest transmission and survival of this bacterial pathogen may be assisted by the free-living protozoa Acanthamoeba. The latter is known to play the role of a host for various pathogenic bacteria, protecting them from harsh environmental conditions. Importantly, there is a similarity between the mechanisms of bacterial survival within amoebae and macrophages, making the former a convenient tool for the investigation of the survival of pathogenic bacteria in the environment. However, the molecular mechanisms involved in the interaction between Campylobacter and Acanthamoeba are not well understood. Whilst some studies suggest the ability of C. jejuni to survive within the protozoa, the other reports support an extracellular mode of survival only. In this review, we focus on the studies investigating the interaction between Campylobacter and Acanthamoeba, address some reasons for the contradictory results, and discuss possible implications of these results for epidemiology. Additionally, as the molecular mechanisms involved remain unknown, we also suggest possible factors that may be involved in this process. Deciphering the molecular mechanisms of pathogen-protozoa interaction will assist in a better understanding of Campylobacter lifestyle and in the development of novel antibacterial drugs.

  19. Campylobacter spp. as a Foodborne Pathogen: A Review

    Science.gov (United States)

    Silva, Joana; Leite, Daniela; Fernandes, Mariana; Mena, Cristina; Gibbs, Paul Anthony; Teixeira, Paula

    2011-01-01

    Campylobacter is well recognized as the leading cause of bacterial foodborne diarrheal disease worldwide. Symptoms can range from mild to serious infections of the children and the elderly and permanent neurological symptoms. The organism is a cytochrome oxidase positive, microaerophilic, curved Gram-negative rod exhibiting corkscrew motility and is carried in the intestine of many wild and domestic animals, particularly avian species including poultry. Intestinal colonization results in healthy animals as carriers. In contrast with the most recent published reviews that cover specific aspects of Campylobacter/campylobacteriosis, this broad review aims at elucidating and discussing the (i) genus Campylobacter, growth and survival characteristics; (ii) detection, isolation and confirmation of Campylobacter; (iii) campylobacteriosis and presence of virulence factors; and (iv) colonization of poultry and control strategies. PMID:21991264

  20. Safety analysis of a Russian phage cocktail: from metagenomic analysis to oral application in healthy human subjects.

    Science.gov (United States)

    McCallin, Shawna; Alam Sarker, Shafiqul; Barretto, Caroline; Sultana, Shamima; Berger, Bernard; Huq, Sayeda; Krause, Lutz; Bibiloni, Rodrigo; Schmitt, Bertrand; Reuteler, Gloria; Brüssow, Harald

    2013-09-01

    Phage therapy has a long tradition in Eastern Europe, where preparations are comprised of complex phage cocktails whose compositions have not been described. We investigated the composition of a phage cocktail from the Russian pharmaceutical company Microgen targeting Escherichia coli/Proteus infections. Electron microscopy identified six phage types, with numerically T7-like phages dominating over T4-like phages. A metagenomic approach using taxonomical classification, reference mapping and de novo assembly identified 18 distinct phage types, including 7 genera of Podoviridae, 2 established and 2 proposed genera of Myoviridae, and 2 genera of Siphoviridae. De novo assembly yielded 7 contigs greater than 30 kb, including a 147-kb Myovirus genome and a 42-kb genome of a potentially new phage. Bioinformatic analysis did not reveal undesired genes and a small human volunteer trial did not associate adverse effects with oral phage exposure.

  1. Phylogenetic diversity and position of the genus Campylobacter

    Science.gov (United States)

    Lau, P. P.; DeBrunner-Vossbrinck, B.; Dunn, B.; Miotto, K.; MacDonnell, M. T.; Rollins, D. M.; Pillidge, C. J.; Hespell, R. B.; Colwell, R. R.; Sogin, M. L.; hide

    1987-01-01

    RNA sequence analysis has been used to examine the phylogenetic position and structure of the genus Campylobacter. A complete 5S rRNA sequence was determined for two strains of Campylobacter jejuni and extensive partial sequences of the 16S rRNA were obtained for several strains of C. jejuni and Wolinella succinogenes. In addition limited partial sequence data were obtained from the 16S rRNAs of isolates of C. coli, C. laridis, C. fetus, C. fecalis, and C. pyloridis. It was found that W. succinogenes is specifically related to, but not included, in the genus Campylobacter as presently constituted. Within the genus significant diversity was noted. C. jejuni, C. coli and C. laridis are very closely related but the other species are distinctly different from one another. C. pyloridis is without question the most divergent of the Campylobacter isolates examined here and is sufficiently distinct to warrant inclusion in a separate genus. In terms of overall position in bacterial phylogeny, the Campylobacter/Wolinella cluster represents a deep branching most probably located within an expanded version of the Division containing the purple photosynthetic bacteria and their relatives. The Campylobacter/Wolinella cluster is not specifically includable in either the alpha, beta or gamma subdivisions of the purple bacteria.

  2. Phylogenetic diversity and position of the genus Campylobacter

    Science.gov (United States)

    Lau, P. P.; DeBrunner-Vossbrinck, B.; Dunn, B.; Miotto, K.; MacDonnell, M. T.; Rollins, D. M.; Pillidge, C. J.; Hespell, R. B.; Colwell, R. R.; Sogin, M. L.; Fox, G. E.

    1987-01-01

    RNA sequence analysis has been used to examine the phylogenetic position and structure of the genus Campylobacter. A complete 5S rRNA sequence was determined for two strains of Campylobacter jejuni and extensive partial sequences of the 16S rRNA were obtained for several strains of C. jejuni and Wolinella succinogenes. In addition limited partial sequence data were obtained from the 16S rRNAs of isolates of C. coli, C. laridis, C. fetus, C. fecalis, and C. pyloridis. It was found that W. succinogenes is specifically related to, but not included, in the genus Campylobacter as presently constituted. Within the genus significant diversity was noted. C. jejuni, C. coli and C. laridis are very closely related but the other species are distinctly different from one another. C. pyloridis is without question the most divergent of the Campylobacter isolates examined here and is sufficiently distinct to warrant inclusion in a separate genus. In terms of overall position in bacterial phylogeny, the Campylobacter/Wolinella cluster represents a deep branching most probably located within an expanded version of the Division containing the purple photosynthetic bacteria and their relatives. The Campylobacter/Wolinella cluster is not specifically includable in either the alpha, beta or gamma subdivisions of the purple bacteria.

  3. Campylobacter jejuni and Campylobacter coli in Children With Acute Diarrhea in Health Centers of Hamadan, Iran

    Directory of Open Access Journals (Sweden)

    Rastyani

    2015-11-01

    Full Text Available Background Enteritis caused by Campylobacter is considered as the most common acute bacterial diarrhea around the world. In most cases, infection occurs as a result of consuming contaminated water or food, especially raw meat of fowls. Objectives The purpose of the present study was to determine the prevalence and antibiotic resistance of campylobacter species among pediatrics of Hamadan city, Iran. Patients and Methods A total of 120 stool samples from children less than 10 years old were examined from January 2013 to December 2014 in Hamadan, Iran. The samples were incubated in Campy-Thio enrichment medium for 1 - 2 hours and then cultured on a specific medium; after that, the suspected colonies were analyzed for Campylobacter spp. identification by conventional tests. The identified species by biochemical methods were confirmed by polymerase chain reaction (PCR. Antimicrobial susceptibility testing was performed by disk agar diffusion (DAD method. Results Twelve (10% Campylobacter spp. from 120 stool samples were isolated including C. coli and C. jejuni. In the antibiotic susceptibility test, the most frequent resistance was observed to ciprofloxacin 8 (88.8%, followed by 7 (77.7% resistant strains to tetracycline, 7 (77.7% to erythromycin, 6 (66.6% to clindamycin, 5 (55.5% to meropenem, 4 (44.4% to gentamicin, 3 (33.3% to nalidixicacid and only 1 (11.1% to chloramphenicol. Conclusions Campylobacter is responsible for some important clinical problems such as enteritis and is also associated with meningitis and hemolytic-uremic syndrome. It is imperative to monitor the prevalence and antibiotic resistance of Campylobacter spp. as well as other the zoonotic bacteria.

  4. Antimicrobial susceptibility profiles and molecular typing of Campylobacter jejuni and Campylobacter coli isolates from ducks in South Korea.

    Science.gov (United States)

    Wei, Bai; Cha, Se-Yeoun; Kang, Min; Roh, Jae-Hee; Seo, Hye-Suk; Yoon, Ran-Hee; Jang, Hyung-Kwan

    2014-12-01

    Campylobacter is a food-borne zoonotic pathogen that causes human gastroenteritis worldwide. Campylobacter bacteria are commensal in the intestines of many food production animals, including ducks and chickens. The objective of the study was to determine the prevalence of Campylobacter species in domestic ducks, and the agar dilution method was used to determine resistance of the isolates to eight antibiotics. In addition, multilocus sequence typing (MLST) was performed to determine the sequence types (STs) of selected Campylobacter isolates. Between May and September 2012, 58 duck farms were analyzed, and 56 (96.6%) were positive for Campylobacter. Among the isolates, 82.1% were Campylobacter jejuni, 16.1% were C. coli, and one was unidentified by PCR. Of the 46 C. jejuni isolates, 87.0%, 10.9%, and 21.7% were resistant to ciprofloxacin, erythromycin, and azithromycin, respectively. Among the C. coli isolates, all 9 strains were resistant to ampicillin, and 77.8% and 33.3% were resistant to ciprofloxacin and azithromycin, respectively. The majority of the Campylobacter isolates were classified as multidrug resistant. Twenty-eight STs were identified, including 20 STs for C. jejuni and 8 STs for C. coli. The most common clonal complexes in C. jejuni were the ST-21 complex and the ST-45 complex, while the ST-828 complex predominated in C. coli. The majority of isolates were of STs noted in ducks and humans from earlier studies, along with seven STs previously associated only with human disease. These STs overlapped between duck and human isolates, indicating that Campylobacter isolates from ducks should be considered potential sources of human infection.

  5. Phage neutralization by sera of patients receiving phage therapy.

    Science.gov (United States)

    Łusiak-Szelachowska, Marzanna; Zaczek, Maciej; Weber-Dąbrowska, Beata; Międzybrodzki, Ryszard; Kłak, Marlena; Fortuna, Wojciech; Letkiewicz, Sławomir; Rogóż, Paweł; Szufnarowski, Krzysztof; Jończyk-Matysiak, Ewa; Owczarek, Barbara; Górski, Andrzej

    2014-08-01

    The aim of our investigation was to verify whether phage therapy (PT) can induce antiphage antibodies. The antiphage activity was determined in sera from 122 patients from the Phage Therapy Unit in Wrocław with bacterial infections before and during PT, and in sera from 30 healthy volunteers using a neutralization test. Furthermore, levels of antiphage antibodies were investigated in sera of 19 patients receiving staphylococcal phages and sera of 20 healthy volunteers using enzyme-linked immunosorbent assay. The phages were administered orally, locally, orally/locally, intrarectally, or orally/intrarectally. The rate of phage inactivation (K) estimated the level of phages' neutralization by human sera. Low K rates were found in sera of healthy volunteers (K ≤ 1.73). Low K rates were detected before PT (K ≤ 1.64). High antiphage activity of sera K > 18 was observed in 12.3% of examined patients (n = 15) treated with phages locally (n = 13) or locally/orally (n = 2) from 15 to 60 days of PT. High K rates were found in patients treated with some Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus faecalis phages. Low K rates were observed during PT in sera of patients using phages orally (K ≤ 1.04). Increased inactivation of phages by sera of patients receiving PT decreased after therapy. These results suggest that the antiphage activity in patients' sera depends on the route of phage administration and phage type. The induction of antiphage activity of sera during or after PT does not exclude a favorable result of PT.

  6. Importance of Campylobacter jejuni for Food Safety and Public Health

    Directory of Open Access Journals (Sweden)

    Omer Cakmak

    2010-04-01

    Full Text Available Campylobacter spp. are microorganisms that can be found in nature in the entire domestic and wild animal’s intestinal flora including the poultry and the sea animals. Campylobacter can better colonize in the poultry than the other animals. Campylobacter jejuni is an important pathogen among the thermophilic Campylobacter spp. whose growth temperature’s are different than the other Campylobacter spp. and can cause serious gastroenteritis in human beings which in some cases ended up with death. Human beings are generally infected with C. jejuni mainly because of the poultry meat and products and rarely because of the red meat which are contaminated during preparation and serving stages. Inadequate cooking, consumption of poorly chlorinated drinking water or unpasteurized milk are other infection sources of C. jejuni. Campylobacteriosis especially affect children under 5 years of age and reported to be a zoonotic illness that cause acute gastroenteritis in human. In many countries, food sourced C. jejuni infections were reported to occur more frequently than Salmonella spp. infections. In order to avoid Campylobacter infections, it is very important to enforce food security programmes and HACCP like systems during growth, slaughterhouses and point of sales stages. Also adequate cooking of the products, hygiene of the kitchen and personnel are important. [TAF Prev Med Bull 2010; 9(2.000: 157-166

  7. Kip, consument en Campylobacter: infectieziektebestrijding met consumentenvoorlichting

    NARCIS (Netherlands)

    Jonge, de R.; Fischer, A.R.H.; Nauta, M.J.

    2007-01-01

    Pluimveevlees is een belangrijke bron van Campylobacter en Campylobacter is een belangrijke verwekker van gastro-enteritis in Nederland. Maatregelen die gericht zijn op het terugbrengen van het aantal Campylobacter-bacteriën in pluimveevlees, bieden geen garantie op Campylobacter-vrij vlees.

  8. Staphylococcus aureus phage types and their correlation to antibiotic resistance

    Directory of Open Access Journals (Sweden)

    Mehndiratta P

    2010-10-01

    Full Text Available Context: Staphylococcus aureus is one of the most devastating human pathogen. The organism has a differential ability to spread and cause outbreak of infections. Characterization of these strains is important to control the spread of infection in the hospitals as well as in the community. Aim: To identify the currently existing phage groups of Staphylococcus aureus, their prevalence and resistance to antibiotics. Materials and Methods: Study was undertaken on 252 Staphylococcus aureus strains isolated from clinical samples. Strains were phage typed and their resistance to antibiotics was determined following standard microbiological procedures. Statistical Analysis: Chi square test was used to compare the antibiotic susceptibility between methicillin resistant Staph. aureus (MRSA and methicillin sensitive S. aureus (MSSA strains. Results: Prevalence of MRSA and MSSA strains was found to be 29.36% and 70.65% respectively. Of these 17.56% of MRSA and 40.44% of MSSA strains were community acquired. All the MSSA strains belonging to phage type 81 from the community were sensitive to all the antibiotics tested including clindamycin and were resistant to penicillin. Forty five percent strains of phage group III and 39% of non-typable MRSA strains from the hospital were resistant to multiple antibiotics. Conclusion: The study revealed that predominant phage group amongst MRSA strains was phage group III and amongst MSSA from the community was phage group NA (phage type 81. MSSA strains isolated from the community differed significantly from hospital strains in their phage type and antibiotic susceptibility. A good correlation was observed between community acquired strains of phage type 81 and sensitivity to gentamycin and clindamycin.

  9. Development of a multiplex PCR assay for identification of Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis subsp. hyointestinalis, Campylobacter jejuni, Campylobacter lari and Campylobacter upsaliensis

    National Research Council Canada - National Science Library

    Yamazaki-Matsune, Wataru; Taguchi, Masumi; Seto, Kazuko; Kawahara, Ryuji; Kawatsu, Kentaro; Kumeda, Yuko; Kitazato, Miyoshi; Nukina, Masafumi; Misawa, Naoaki; Tsukamoto, Teizo

    2007-01-01

    ...{at}iph.pref.osaka.jp Received 26 April 2007 Accepted 9 July 2007 A multiplex PCR assay has been developed for the identification of the six common Campylobacter taxa associated with human gastroenteritis...

  10. Campylobacter hyointestinalis subsp hyointestinalis, a common Campylobacter species in reindeer

    DEFF Research Database (Denmark)

    Hanninen, M.L.; Sarelli, L.; Sukura, A.

    2002-01-01

    Aims: To study the prevalence of Campylobacter spp. in the faecal material of reindeer, and to identify the isolates by means of a polyphasic approach. In addition, to study the genetic diversity of Camp. hyointestinalis subsp. hyointestinalis reindeer isolates by pulsed-field gel electrophoresis...... slaughterhouses. Samples were cultured by methods suitable for isolation of fastidious Campylobacter species. Of all samples, 6% (24/399) were Campylobacter-positive. Phenotypic characteristics, SDS-PAGE protein patterns, dot blot DNA-DNA hybridization, 23S rDNA restriction fragment polymorphism analysis and PFGE...... identified the isolates as Camp. hyointestinalis subsp. kyointestinalis. Conclusions: Campylobacter hyointestinalis subsp. hyointestinalis was the only Campylobacter species isolated from reindeer in this study. The isolates showed high genomic diversity in PFGE with the restriction enzymes SmaI and Kpn...

  11. Phage “delay” towards enhancing bacterial escape from biofilms: a more comprehensive way of viewing resistance to bacteriophages

    Directory of Open Access Journals (Sweden)

    Stephen T. Abedon

    2017-03-01

    Full Text Available In exploring bacterial resistance to bacteriophages, emphasis typically is placed on those mechanisms which completely prevent phage replication. Such resistance can be detected as extensive reductions in phage ability to form plaques, that is, reduced efficiency of plating. Mechanisms include restriction-modification systems, CRISPR/Cas systems, and abortive infection systems. Alternatively, phages may be reduced in their “vigor” when infecting certain bacterial hosts, that is, with phages displaying smaller burst sizes or extended latent periods rather than being outright inactivated. It is well known, as well, that most phages poorly infect bacteria that are less metabolically active. Extracellular polymers such as biofilm matrix material also may at least slow phage penetration to bacterial surfaces. Here I suggest that such “less-robust” mechanisms of resistance to bacteriophages could serve bacteria by slowing phage propagation within bacterial biofilms, that is, delaying phage impact on multiple bacteria rather than necessarily outright preventing such impact. Related bacteria, ones that are relatively near to infected bacteria, e.g., roughly 10+ µm away, consequently may be able to escape from biofilms with greater likelihood via standard dissemination-initiating mechanisms including erosion from biofilm surfaces or seeding dispersal/central hollowing. That is, given localized areas of phage infection, so long as phage spread can be reduced in rate from initial points of contact with susceptible bacteria, then bacterial survival may be enhanced due to bacteria metaphorically “running away” to more phage-free locations. Delay mechanisms—to the extent that they are less specific in terms of what phages are targeted—collectively could represent broader bacterial strategies of phage resistance versus outright phage killing, the latter especially as require specific, evolved molecular recognition of phage presence. The

  12. Safety and efficacy of phage therapy via the intravenous route.

    Science.gov (United States)

    Speck, Peter; Smithyman, Anthony

    2016-02-01

    Increasing development of antimicrobial resistance is driving a resurgence in interest in phage therapy: the use of bacteriophages to treat bacterial infections. As the lytic action of bacteriophages is unaffected by the antibiotic resistance status of their bacterial target, it is thought that phage therapy may have considerable potential in the treatment of a wide range of topical and localized infections. As yet this interest has not extended to intravenous (IV) use, which is surprising given that the historical record shows that phages are likely to be safe and effective when delivered by this route. Starting almost 100 years ago, phages were administered intravenously in treatment of systemic infections including typhoid, and Staphylococcal bacteremia. There was extensive IV use of phages in the 1940s to treat typhoid, reportedly with outstanding efficacy and safety. The safety of IV phage administration is also underpinned by the detailed work of Ochs and colleagues in Seattle who have over four decades' experience with IV injection into human subjects of large doses of highly purified coliphage PhiX174. Though these subjects included a large number of immune-deficient children, no serious side effects were observed over this extended time period. The large and continuing global health problems of typhoid and Staphylococcus aureus are exacerbated by the increasing antibiotic resistance of these pathogens. We contend that these infections are excellent candidates for use of IV phage therapy.

  13. Detection of Campylobacter jejuni in raw meat

    OpenAIRE

    Z Noori; SH Saadati; A. Mirsalehian; SH Shoeibi; N Rahimifard; Mehrangiz Mehdizadeh; M Pirali- Hamedani

    2009-01-01

    Background & Objectives: Campylobacter jejuni is a Gram negative, microaerophilic, non-spore-forming and a small"ncurved bacillus which is able to cause foodborne infection in human. In this study the occurrence of C. jejuni in poultry and"nbeef meat was investigated."nMaterials & Methods: Forty raw meat samples including 22 poultry samples and 18 beef samples were investigated for the"npresence of C. jejuni. To isolate the bacterium, the samples were initially enriched in Preston Bro...

  14. PRESENCE OF RESISTANCE IN CAMPYLOBACTER JEJUNI AND CAMPYLOBACTER COLI

    Directory of Open Access Journals (Sweden)

    Branislava Kocić

    2009-04-01

    Full Text Available There are 18 species belonging to the genus of Campylobacter (rRNK group I, of which thermophilic ones are the following: Campylobacter jejuni, Campylobacter coli, Campylobacter lari and Campylobacter upsaliensis. The aim of our research was to determine the sensitivity of Campylobacter species, isolated from human feces, to antibiotics being used in practice. The study involved 50 human strains of C. jejuni/coli isolated from feces in the Center for Microbiology in the Public Health Institute Nis. Sensitivity was tested by applying the disk diffusion method on seven antibiotics (erythromycin, gentamicin, tetracycline, ciprofloxacin, hloramphenicol, cephalexin and nalidixic acid. Our results showed low resistance to erythromycin, gentamicin and tetracycline (2%, which corresponds to the studies conducted in the world. Moreover, these findings indicate that erythromycin may be considered the drug of choice in the treatment of Campylobacter diarrhea in this region. Resistance to fluoroquinolone and nalidixic acid was 44%, and C. coli showed higher resistance compared to C. jejuni, though statistical significance was not proved.

  15. Phage cocktails and the future of phage therapy.

    Science.gov (United States)

    Chan, Benjamin K; Abedon, Stephen T; Loc-Carrillo, Catherine

    2013-06-01

    Viruses of bacteria, known as bacteriophages or phages, were discovered nearly 100 years ago. Their potential as antibacterial agents was appreciated almost immediately, with the first 'phage therapy' trials predating Fleming's discovery of penicillin by approximately a decade. In this review, we consider phage therapy that can be used for treating bacterial infections in humans, domestic animals and even biocontrol in foods. Following an overview of the topic, we explore the common practice - both experimental and, in certain regions of the world, clinical - of mixing therapeutic phages into cocktails consisting of multiple virus types. We conclude with a discussion of the commercial and medical context of phage cocktails as therapeutic agents. In comparing off-the-shelf versus custom approaches, we consider the merits of a middle ground, which we deem 'modifiable'. Finally, we explore a regulatory framework for such an approach based on an influenza vaccine model.

  16. Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine

    Science.gov (United States)

    Nobrega, Franklin L.; Costa, Ana Rita; Santos, José F.; Siliakus, Melvin F.; van Lent, Jan W. M.; Kengen, Servé W. M.; Azeredo, Joana; Kluskens, Leon D.

    2016-01-01

    Orally administered phages to control zoonotic pathogens face important challenges, mainly related to the hostile conditions found in the gastrointestinal tract (GIT). These include temperature, salinity and primarily pH, which is exceptionally low in certain compartments. Phage survival under these conditions can be jeopardized and undermine treatment. Strategies like encapsulation have been attempted with relative success, but are typically complex and require several optimization steps. Here we report a simple and efficient alternative, consisting in the genetic engineering of phages to display lipids on their surfaces. Escherichia coli phage T7 was used as a model and the E. coli PhoE signal peptide was genetically fused to its major capsid protein (10 A), enabling phospholipid attachment to the phage capsid. The presence of phospholipids on the mutant phages was confirmed by High Performance Thin Layer Chromatography, Dynamic Light Scattering and phospholipase assays. The stability of phages was analysed in simulated GIT conditions, demonstrating improved stability of the mutant phages with survival rates 102–107 pfu.mL−1 higher than wild-type phages. Our work demonstrates that phage engineering can be a good strategy to improve phage tolerance to GIT conditions, having promising application for oral administration in veterinary medicine. PMID:27976713

  17. Biofilm control with natural and genetically-modified phages.

    Science.gov (United States)

    Motlagh, Amir Mohaghegh; Bhattacharjee, Ananda Shankar; Goel, Ramesh

    2016-04-01

    Bacteriophages, as the most dominant and diverse entities in the universe, have the potential to be one of the most promising therapeutic agents. The emergence of multidrug-resistant bacteria and the antibiotic crisis in the last few decades have resulted in a renewed interest in phage therapy. Furthermore, bacteriophages, with the capacity to rapidly infect and overcome bacterial resistance, have demonstrated a sustainable approach against bacterial pathogens-particularly in biofilm. Biofilm, as complex microbial communities located at interphases embedded in a matrix of bacterial extracellular polysaccharide substances (EPS), is involved in health issues such as infections associated with the use of biomaterials and chronic infections by multidrug resistant bacteria, as well as industrial issues such as biofilm formation on stainless steel surfaces in food industry and membrane biofouling in water and wastewater treatment processes. In this paper, the most recent studies on the potential of phage therapy using natural and genetically-modified lytic phages and their associated enzymes in fighting biofilm development in various fields including engineering, industry, and medical applications are reviewed. Phage-mediated prevention approaches as an indirect phage therapy strategy are also explored in this review. In addition, the limitations of these approaches and suggestions to overcome these constraints are discussed to enhance the efficiency of phage therapy process. Finally, future perspectives and directions for further research towards a better understanding of phage therapy to control biofilm are recommended.

  18. Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar.

    Science.gov (United States)

    Gonsalves, Camila Cristina; Borsoi, Anderlise; Perdoncini, Gustavo; Rodrigues, Laura Beatriz; do Nascimento, Vladimir Pinheiro

    2016-01-01

    Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  19. Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar

    Directory of Open Access Journals (Sweden)

    Camila Cristina Gonsalves

    Full Text Available ABSTRACT Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples. Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products.

  20. The epic of phage therapy

    OpenAIRE

    Alain Dublanchet; Shawna Bourne

    2007-01-01

    The present report describes the presentation given by Dr Alain Dublanchet at the Stanier/Oxford Hygiene Symposium, held in Oxford, England, on November 10, 2004. Dr Dublanchet's lecture, entitled ‘The epic of phage therapy’, provided a sequential account of the use of phage as an antimicrobial from its discovery to its rise and fall and current rediscovery.

  1. Interference with phage lambda development by the small subunit of the phage 21 terminase, gp1.

    OpenAIRE

    1991-01-01

    Bacteriophage lambda development is blocked in cells carrying a plasmid that expresses the terminase genes of phage 21. The interference is caused by the small subunit of phage 21 terminase, gp1. Mutants of lambda able to form plaques in the presence of gp1 include sti mutants. One such mutation, sti30, is an A. T-to-G.C transition mutation at base pair 184 on the lambda chromosome. The sti30 mutation extends the length of the ribosome-binding sequence of the Nul gene that is complementary to...

  2. Clinical relevance of infections with zoonotic and human oral species of Campylobacter.

    Science.gov (United States)

    Lee, Soomin; Lee, Jeeyeon; Ha, Jimyeong; Choi, Yukyung; Kim, Sejeong; Lee, Heeyoung; Yoon, Yohan; Choi, Kyoung-Hee

    2016-07-01

    Genus Campylobacter has been recognized as a causative bacterial agent of animal and human diseases. Human Campylobacter infections have caused more concern. Campylobacters can be classified into two groups in terms of their original host: zoonotic and human oral species. The major zoonotic species are Campylobacter jejuni and Campylobacter coli, which mostly reside in the intestines of avian species and are transmitted to humans via consumption of contaminated poultry products, thus causing human gastroenteritis and other diseases as sequelae. The other campylobacters, human oral species, include C. concisus, C. showae, C. gracilis, C. ureolyticus, C. curvus, and C. rectus. These species are isolated from the oral cavity, natural colonization site, but have potential clinical relevance in the periodontal region to varying extent. Two species, C. jejuni and C. coli, are believed to be mainly associated with intestinal diseases, but recent studies suggested that oral Campylobacter species also play a significant role in intestinal diseases. This review offers an outline of the two Campylobacter groups (zoonotic and human oral), their virulence traits, and the associated illnesses including gastroenteritis.

  3. Detection of Campylobacter jejuni in raw meat

    Directory of Open Access Journals (Sweden)

    Z Noori

    2009-12-01

    Full Text Available Background & Objectives: Campylobacter jejuni is a Gram negative, microaerophilic, non-spore-forming and a small"ncurved bacillus which is able to cause foodborne infection in human. In this study the occurrence of C. jejuni in poultry and"nbeef meat was investigated."nMaterials & Methods: Forty raw meat samples including 22 poultry samples and 18 beef samples were investigated for the"npresence of C. jejuni. To isolate the bacterium, the samples were initially enriched in Preston Broth medium and subsequently"ntransferred to Campylobacter selective Agar containing defibrinated sheep blood and antibiotics. The biochemical tests were"nused for identification of isolated bacteria at species level."nResults: Three poultry samples were positive for C. jejuni."nConclusion: Alimentary tract of chickens contain high numbers of C. jejuni therefore, this bacterium can be easily found in"ntheir feces. It is recommended to use chlorinated water in birds’ feed and to perform slaughtering, skinning and evisceration"nunder aseptic conditions to prevent campylobacteriosis in human from poultry meat. None of the beef samples yielded any"nCampylobacter, this may be due to limited number of samples of beef meat analyzed in this study.

  4. Current and Potential Treatments for Reducing Campylobacter Colonization in Animal Hosts and Disease in Humans

    Science.gov (United States)

    Johnson, Tylor J.; Shank, Janette M.; Johnson, Jeremiah G.

    2017-01-01

    Campylobacter jejuni is the leading cause of bacteria-derived gastroenteritis worldwide. In the developed world, Campylobacter is usually acquired by consuming under-cooked poultry, while in the developing world it is often obtained through drinking contaminated water. Once consumed, the bacteria adhere to the intestinal epithelium or mucus layer, causing toxin-mediated inhibition of fluid reabsorption from the intestine and invasion-induced inflammation and diarrhea. Traditionally, severe or prolonged cases of campylobacteriosis have been treated with antibiotics; however, overuse of these antibiotics has led to the emergence of antibiotic-resistant strains. As the incidence of antibiotic resistance, emergence of post-infectious diseases, and economic burden associated with Campylobacter increases, it is becoming urgent that novel treatments are developed to reduce Campylobacter numbers in commercial poultry and campylobacteriosis in humans. The purpose of this review is to provide the current status of present and proposed treatments to combat Campylobacter infection in humans and colonization in animal reservoirs. These treatments include anti-Campylobacter compounds, probiotics, bacteriophage, vaccines, and anti-Campylobacter bacteriocins, all of which may be successful at reducing the incidence of campylobacteriosis in humans and/or colonization loads in poultry. In addition to reviewing treatments, we will also address several proposed targets that may be used in future development of novel anti-Campylobacter treatments. PMID:28386253

  5. Phages targeting infected tissues: novel approach to phage therapy.

    Science.gov (United States)

    Górski, Andrzej; Dąbrowska, Krystyna; Hodyra-Stefaniak, Katarzyna; Borysowski, Jan; Międzybrodzki, Ryszard; Weber-Dąbrowska, Beata

    2015-01-01

    While the true efficacy of phage therapy still requires formal confirmation in clinical trials, it continues to offer realistic potential treatment in patients in whom antibiotics have failed. Novel developments and approaches are therefore needed to ascertain that future clinical trials would evaluate the therapy in its optimal form thus allowing for reliable conclusions regarding the true value of phage therapy. In this article, we present our vision to develop and establish a bank of phages specific to most threatening pathogens and armed with homing peptides enabling their localization in infected tissues in densities assuring efficient and stable eradication of infection.

  6. The use of probabilistic graphical models (PGMs) to develop a cost-effective vaccination strategy against Campylobacter in poultry

    DEFF Research Database (Denmark)

    Garcia Clavero, Ana Belén; Madsen, A.; Vigre, Håkan

    , epidemiological and economic factors (cost-reward functions) have been included in the models. The final outcome of the models is presented in probabilities of expected level of Campylobacter and financial terms influenced by the decision on vaccination. For example, if the best decision seems to be to vaccinate......Human campylobacteriosis represents an important economic and public health problem. Campylobacter originating from feces of infected chickens will contaminate chicken meat posing a risk to the consumer. Vaccination against Campylobacter in broilers is one probable measure to reduce consumers......’ exposure to Campylobacter.In this presentation we focus on the development of a computerized decision support system to aid management decisions on Campylobacter vaccination of commercial broilers. Broilers should be vaccinated against Campylobacter in the first 2 weeks of age. Therefore, the decision...

  7. Comparative genomic and morphological analyses of Listeria phages isolated from farm environments.

    Science.gov (United States)

    Denes, Thomas; Vongkamjan, Kitiya; Ackermann, Hans-Wolfgang; Moreno Switt, Andrea I; Wiedmann, Martin; den Bakker, Henk C

    2014-08-01

    The genus Listeria is ubiquitous in the environment and includes the globally important food-borne pathogen Listeria monocytogenes. While the genomic diversity of Listeria has been well studied, considerably less is known about the genomic and morphological diversity of Listeria bacteriophages. In this study, we sequenced and analyzed the genomes of 14 Listeria phages isolated mostly from New York dairy farm environments as well as one related Enterococcus faecalis phage to obtain information on genome characteristics and diversity. We also examined 12 of the phages by electron microscopy to characterize their morphology. These Listeria phages, based on gene orthology and morphology, together with previously sequenced Listeria phages could be classified into five orthoclusters, including one novel orthocluster. One orthocluster (orthocluster I) consists of large genome (~135-kb) myoviruses belonging to the genus “Twort-like viruses,” three orthoclusters (orthoclusters II to IV) contain small-genome (36- to 43-kb) siphoviruses with icosahedral heads, and the novel orthocluster V contains medium-sized-genome (~66-kb) siphoviruses with elongated heads. A novel orthocluster (orthocluster VI) of E. faecalis phages, with medium-sized genomes (~56 kb), was identified, which grouped together and shares morphological features with the novel Listeria phage orthocluster V. This new group of phages (i.e., orthoclusters V and VI) is composed of putative lytic phages that may prove to be useful in phage-based applications for biocontrol, detection, and therapeutic purposes.

  8. Survival and resuscitation of ten strains of Campylobacter jejuni and Campylobacter coli under acid conditions

    NARCIS (Netherlands)

    Chaveerach, P.; Huurne, ter A.A.H.M.; Lipman, L.J.A.; Knapen, van F.

    2003-01-01

    The culturability of 10 strains of Campylobacter jejuni and Campylobacter coli was studied after the bacteria were exposed to acid conditions for various periods of time. Campylobacter cells could not survive 2 h under acid conditions (formic acid at pH 4). The 10 Campylobacter strains could not be

  9. Survival and resuscitation of ten strains of Campylobacter jejuni and Campylobacter coli under acid conditions

    NARCIS (Netherlands)

    Chaveerach, P.; Huurne, ter A.A.H.M.; Lipman, L.J.A.; Knapen, van F.

    2003-01-01

    The culturability of 10 strains of Campylobacter jejuni and Campylobacter coli was studied after the bacteria were exposed to acid conditions for various periods of time. Campylobacter cells could not survive 2 h under acid conditions (formic acid at pH 4). The 10 Campylobacter strains could not be

  10. Persistence of bacteria and phages in a chemostat.

    Science.gov (United States)

    Smith, Hal L; Thieme, Horst R

    2012-05-01

    The model of bacteriophage predation on bacteria in a chemostat formulated by Levin et al. (Am Nat 111:3-24, 1977) is generalized to include a distributed latent period, distributed viral progeny release from infected bacteria, unproductive adsorption of phages to infected cells, and possible nutrient uptake by infected cells. Indeed, two formulations of the model are given: a system of delay differential equations with infinite delay, and a more general infection-age model that leads to a system of integro-differential equations. It is shown that the bacteria persist, and sharp conditions for persistence and extinction of phages are determined by the reproductive ratio for phage relative to the phage-free equilibrium. A novel feature of our analysis is the use of the Laplace transform.

  11. Advances in Campylobacter biology and implications for biotechnological applications

    Science.gov (United States)

    Jeon, Byeonghwa; Muraoka, Wayne T.; Zhang, Qijing

    2010-01-01

    Summary Campylobacter jejuni is a major foodborne pathogen of animal origin and a leading cause of bacterial gastroenteritis in humans. During the past decade, especially since the publication of the first C. jejuni genome sequence, major advances have been made in understanding the pathobiology and physiology of this organism. It is apparent that C. jejuni utilizes sophisticated mechanisms for effective colonization of the intestinal tracts in various animal species. Although Campylobacter is fragile in the environment and requires fastidious growth conditions, it exhibits great flexibility in the adaptation to various habitats including the gastrointestinal tract. This high adaptability is attributable to its genetically, metabolically and phenotypically diverse population structure and its ability to change in response to various challenges. Unlike other enteric pathogens, such as Escherichia coli and Salmonella, Campylobacter is unable to utilize exogenous glucose and mainly depends on the catabolism of amino acids as a carbon source. Campylobacter proves highly mutable in response to antibiotic treatments and possesses eukaryote‐like dual protein glycosylation systems, which modify flagella and other surface proteins with specific sugar structures. In this review we will summarize the distinct biological traits of Campylobacter and discuss the potential biotechnological approaches that can be developed to control this enteric pathogen. PMID:21255325

  12. [Campylobacter and Salmonella acute gastroenteritis: epidemiology and health care utilization].

    Science.gov (United States)

    Sala Farré, Maria Rosa; Osorio Sánchez, Dimelza; Arias Varela, Cesar; Simó Sanahuja, Maria; Recasens Recasens, Assumpta; Pérez Jové, Josefa

    2015-10-05

    In Catalonia the current surveillance systems do not allow to know the true incidence or the health care utilization of acute gastroenteritis (AGE) caused by Campylobacter and Salmonella infections. The aim of this study is to analyze these characteristics. Descriptive study of Campylobacter and Salmonella infections reported in 2002 and 2012 in Catalonia, Spain. We included cases isolated and reported by the laboratory to a regional Surveillance Unit. The estimated incidence of Salmonella and Campylobacter AGE decreased by almost 50% and 20% respectively in 2012. Children between one and 4 years old were the most affected in both years. Significant differences in the clinical characteristics and disease duration were observed between Campylobacter and Salmonella. Visits to the Emergency Department and hospitalization rates were 63.7% and 15%, being more frequent among salmonellosis cases. The estimated incidence of Campylobacter and Salmonella infections has decreased, however rates are still important, as well as it is the health care utilization in both diseases. Current surveillance systems need appropriateness improvements to reach a better control of these infections. Copyright © 2014 Elsevier España, S.L.U. All rights reserved.

  13. Information Phage Therapy Research Should Report

    OpenAIRE

    Stephen T. Abedon

    2017-01-01

    Bacteriophages, or phages, are viruses which infect bacteria. A large subset of phages infect bactericidally and, consequently, for nearly one hundred years have been employed as antibacterial agents both within and outside of medicine. Clinically these applications are described as phage or bacteriophage therapy. Alternatively, and especially in the treatment of environments, this practice instead may be described as a phage-mediated biocontrol of bacteria. Though the history of phage therap...

  14. Antimicrobial Resistance Mechanisms among Campylobacter

    Science.gov (United States)

    2013-01-01

    Campylobacter jejuni and Campylobacter coli are recognized as the most common causative agents of bacterial gastroenteritis in the world. Humans most often become infected by ingesting contaminated food, especially undercooked chicken, but also other sources of bacteria have been described. Campylobacteriosis is normally a self-limiting disease. Antimicrobial treatment is needed only in patients with more severe disease and in those who are immunologically compromised. The most common antimicrobial agents used in the treatment of Campylobacter infections are macrolides, such as erythromycin, and fluoroquinolones, such as ciprofloxacin. Tetracyclines have been suggested as an alternative choice in the treatment of clinical campylobacteriosis but in practice are not often used. However, during the past few decades an increasing number of resistant Campylobacter isolates have developed resistance to fluoroquinolones and other antimicrobials such as macrolides, aminoglycosides, and beta-lactams. Trends in antimicrobial resistance have shown a clear correlation between use of antibiotics in the veterinary medicine and animal production and resistant isolates of Campylobacter in humans. In this review, the patterns of emerging resistance to the antimicrobial agents useful in treatment of the disease are presented and the mechanisms of resistance to these drugs in Campylobacter are discussed. PMID:23865047

  15. Antimicrobial resistance mechanisms among Campylobacter.

    Science.gov (United States)

    Wieczorek, Kinga; Osek, Jacek

    2013-01-01

    Campylobacter jejuni and Campylobacter coli are recognized as the most common causative agents of bacterial gastroenteritis in the world. Humans most often become infected by ingesting contaminated food, especially undercooked chicken, but also other sources of bacteria have been described. Campylobacteriosis is normally a self-limiting disease. Antimicrobial treatment is needed only in patients with more severe disease and in those who are immunologically compromised. The most common antimicrobial agents used in the treatment of Campylobacter infections are macrolides, such as erythromycin, and fluoroquinolones, such as ciprofloxacin. Tetracyclines have been suggested as an alternative choice in the treatment of clinical campylobacteriosis but in practice are not often used. However, during the past few decades an increasing number of resistant Campylobacter isolates have developed resistance to fluoroquinolones and other antimicrobials such as macrolides, aminoglycosides, and beta-lactams. Trends in antimicrobial resistance have shown a clear correlation between use of antibiotics in the veterinary medicine and animal production and resistant isolates of Campylobacter in humans. In this review, the patterns of emerging resistance to the antimicrobial agents useful in treatment of the disease are presented and the mechanisms of resistance to these drugs in Campylobacter are discussed.

  16. Phage sensitivity and prophage carriage in Staphylococcus aureus isolated from foods in Spain and New Zealand.

    Science.gov (United States)

    Gutiérrez, Diana; Rodríguez-Rubio, Lorena; García, Pilar; Billington, Craig; Premarante, Aruni; Rodríguez, Ana; Martínez, Beatriz

    2016-08-02

    Bacteriophages (phages) are a promising tool for the biocontrol of pathogenic bacteria, including those contaminating food products and causing infectious diseases. However, the success of phage preparations is limited by the host ranges of their constituent phages. The phage resistance/sensitivity profile of eighty seven Staphylococcus aureus strains isolated in Spain and New Zealand from dairy, meat and seafood sources was determined for six phages (Φ11, K, ΦH5, ΦA72, CAPSa1 and CAPSa3). Most of the S. aureus strains were sensitive to phage K (Myoviridae) and CAPSa1 (Siphoviridae) regardless of their origin. There was a higher sensitivity of New Zealand S. aureus strains to phages isolated from both Spain (ΦH5 and ΦA72) and New Zealand (CAPSa1 and CAPSa3). Spanish phages had a higher infectivity on S. aureus strains of Spanish dairy origin, while Spanish strains isolated from other environments were more sensitive to New Zealand phages. Lysogeny was more prevalent in Spanish S. aureus compared to New Zealand strains. A multiplex PCR reaction, which detected ΦH5 and ΦA72 sequences, indicated a high prevalence of these prophages in Spanish S. aureus strains, but were infrequently detected in New Zealand strains. Overall, the correlation between phage resistance and lysogeny in S. aureus strains was found to be weak.

  17. Phage encoded H-NS: a potential achilles heel in the bacterial defence system.

    Directory of Open Access Journals (Sweden)

    Connor T Skennerton

    Full Text Available The relationship between phage and their microbial hosts is difficult to elucidate in complex natural ecosystems. Engineered systems performing enhanced biological phosphorus removal (EBPR, offer stable, lower complexity communities for studying phage-host interactions. Here, metagenomic data from an EBPR reactor dominated by Candidatus Accumulibacter phosphatis (CAP, led to the recovery of three complete and six partial phage genomes. Heat-stable nucleoid structuring (H-NS protein, a global transcriptional repressor in bacteria, was identified in one of the complete phage genomes (EPV1, and was most similar to a homolog in CAP. We infer that EPV1 is a CAP-specific phage and has the potential to repress up to 6% of host genes based on the presence of putative H-NS binding sites in the CAP genome. These genes include CRISPR associated proteins and a Type III restriction-modification system, which are key host defense mechanisms against phage infection. Further, EPV1 was the only member of the phage community found in an EBPR microbial metagenome collected seven months prior. We propose that EPV1 laterally acquired H-NS from CAP providing it with a means to reduce bacterial defenses, a selective advantage over other phage in the EBPR system. Phage encoded H-NS could constitute a previously unrecognized weapon in the phage-host arms race.

  18. Phage Encoded H-NS: A Potential Achilles Heel in the Bacterial Defence System

    Science.gov (United States)

    Skennerton, Connor T.; Angly, Florent E.; Breitbart, Mya; Bragg, Lauren; He, Shaomei; McMahon, Katherine D.; Hugenholtz, Philip; Tyson, Gene W.

    2011-01-01

    The relationship between phage and their microbial hosts is difficult to elucidate in complex natural ecosystems. Engineered systems performing enhanced biological phosphorus removal (EBPR), offer stable, lower complexity communities for studying phage-host interactions. Here, metagenomic data from an EBPR reactor dominated by Candidatus Accumulibacter phosphatis (CAP), led to the recovery of three complete and six partial phage genomes. Heat-stable nucleoid structuring (H-NS) protein, a global transcriptional repressor in bacteria, was identified in one of the complete phage genomes (EPV1), and was most similar to a homolog in CAP. We infer that EPV1 is a CAP-specific phage and has the potential to repress up to 6% of host genes based on the presence of putative H-NS binding sites in the CAP genome. These genes include CRISPR associated proteins and a Type III restriction-modification system, which are key host defense mechanisms against phage infection. Further, EPV1 was the only member of the phage community found in an EBPR microbial metagenome collected seven months prior. We propose that EPV1 laterally acquired H-NS from CAP providing it with a means to reduce bacterial defenses, a selective advantage over other phage in the EBPR system. Phage encoded H-NS could constitute a previously unrecognized weapon in the phage-host arms race. PMID:21625595

  19. Aspects of epidemiology of Campylobacter in poultry

    NARCIS (Netherlands)

    Jacobs-Reitsma, W.F.

    1997-01-01

    Campylobacter bacteria, which in humans cause infections with severe symptoms of diarrhoea, are mainly transmitted by food, especially poultry meat products. Several studies on Campylobacter colonization in breeders, laying hens, and broilers were carried out. Isolates were serotyped, using a

  20. Campylobacter as a venereal disease in cattle

    NARCIS (Netherlands)

    Dijkstra, T.; Wagenaar, J.A.; Visser, I.J.; Bergen, van M.A.P.; Pastoor, P.W.; Strampel, J.; Kock, P.A.

    2005-01-01

    Since the introduction of AI, venereal diseases caused by Tritrichomonas fetus and Campylobacter fetus subsp. venerealis haved been eradicated in The Netherlands. Campylobacter fetus subsp. fetus can cause sporadic abortion and early embryonic death. When natural breeding is practised, venereal

  1. Methicillin-resistant Staphylococcus aureus phage plaque size enhancement using sublethal concentrations of antibiotics.

    Science.gov (United States)

    Kaur, Sandeep; Harjai, Kusum; Chhibber, Sanjay

    2012-12-01

    Phage therapy presents an alternative approach against the emerging methicillin-resistant Staphylococcus aureus (MRSA) threat. Some of the problems encountered during isolation of MRSA phages include the high prevalence of enteric phages in natural sources, nonspecific absorption of viable phage, and the formation of pinpoint or tiny plaques. The phage isolated in this study, MR-5, also formed tiny plaques against its host S. aureus ATCC 43300 (MRSA), making its detection and enumeration difficult. An improved method of increasing the plaque size of MRSA phage by incorporating sublethal concentrations of three different classes of antibiotics (inhibitors of protein synthesis) in the classical double-layer agar (DLA) method was investigated. The β-lactam and quinolone antibiotics commonly employed in earlier studies for increasing the plaque size did not show any significant effect on the plaque size of isolated MR-5 phage. Linezolid (oxazolidinone class), tetracycline, and ketolide antibiotics brought significant enhancements (3 times the original size) in the plaque size of MR-5 phage. Prior treatment with these antibiotics resulted in significant reductions in the time of adsorption and the latent period of MR-5 phage. To rule out whether the action of linezolid (which brought the maximum increase in plaque size) was specific for a single phage only, its effect on the plaque size of seven other S. aureus-specific phages was also assessed. Significant enhancements in the plaque size of these phages were observed. These results indicate that this modification can therefore safely be incorporated in the traditional DLA overlay method to search for new MRSA-virulent phages.

  2. Properties of Klebsiella Phage P13 and Associated Exopolysaccharide Depolymerase

    Institute of Scientific and Technical Information of China (English)

    LIU Yang; LI Guiyang; MO Zhaolan; CHAI Zihan; SHANG Anqi; MOU Haijin

    2014-01-01

    The bacteriophage P13 that infects Klebsiella serotype K13 contains a heat-stable depolymerase capable of effective degradation of exopolysaccharide (EPS) produced by this microorganism. In this study, the titer of phage P13, initially 2.0 × 107 pfu mL-1, was found increasing 20 min after infection and reached 5.0 × 109 pfu mL-1 in 60 min. Accordingly, the enzyme activity of de-polymerase approached the maximum 60 min after infection. Treatment at 70℃for 30 min inactivated all the phage, but retained over 90%of the depolymerase activity. Addition of acetone into the crude phage lysate led to precipitation of the protein, with a marked increase in bacterial EPS degradation activity and a rapid drop in the titer of phage. After partial purification by acetone precipitation and ultrafiltration centrifugation, the enzyme was separated from the phage particles, showing two components with enzyme activity on Q-Sepharose Fast Flow. The soluble enzyme had an optimum degradation activity at 60℃and pH 6.5. Transmission electron mi-croscopy demonstrated that the phage P13 particles were spherical with a diameter of 50 nm and a short stumpy tail. It was a dou-ble-strand DNA virus consisting of a nucleic acid molecule of 45976 bp. This work provides an efficient purification operation in-cluding thermal treatment and ultrafiltration centrifugation, to dissociate depolymerase from phage particles. The characterization of phage P13 and associated EPS depolymerase is beneficial for further application of this enzyme.

  3. Phage therapy against Enterococcus faecalis in dental root canals

    Directory of Open Access Journals (Sweden)

    Leron Khalifa

    2016-09-01

    Full Text Available Antibiotic resistance is an ever-growing problem faced by all major sectors of health care, including dentistry. Recurrent infections related to multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus, carbapenem-resistant Enterobacteriaceae, and vancomycin-resistant enterococci (VRE in hospitals are untreatable and question the effectiveness of notable drugs. Two major reasons for these recurrent infections are acquired antibiotic resistance genes and biofilm formation. None of the traditionally known effective techniques have been able to efficiently resolve these issues. Hence, development of a highly effective antibacterial practice has become inevitable. One example of a hard-to-eradicate pathogen in dentistry is Enterococcus faecalis, which is one of the most common threats observed in recurrent root canal treatment failures, of which the most problematic to treat are its biofilm-forming VRE strains. An effective response against such infections could be the use of bacteriophages (phages. Phage therapy was found to be highly effective against biofilm and multidrug-resistant bacteria and has other advantages like ease of isolation and possibilities for genetic manipulations. The potential of phage therapy in dentistry, in particular against E. faecalis biofilms in root canals, is almost unexplored. Here we review the efforts to develop phage therapy against biofilms. We also focus on the phages isolated against E. faecalis and discuss the possibility of using phages against E. faecalis biofilm in root canals.

  4. Phage therapy against Enterococcus faecalis in dental root canals

    Science.gov (United States)

    Khalifa, Leron; Shlezinger, Mor; Beyth, Shaul; Houri-Haddad, Yael; Coppenhagen-Glazer, Shunit; Beyth, Nurit; Hazan, Ronen

    2016-01-01

    Antibiotic resistance is an ever-growing problem faced by all major sectors of health care, including dentistry. Recurrent infections related to multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus, carbapenem-resistant Enterobacteriaceae, and vancomycin-resistant enterococci (VRE) in hospitals are untreatable and question the effectiveness of notable drugs. Two major reasons for these recurrent infections are acquired antibiotic resistance genes and biofilm formation. None of the traditionally known effective techniques have been able to efficiently resolve these issues. Hence, development of a highly effective antibacterial practice has become inevitable. One example of a hard-to-eradicate pathogen in dentistry is Enterococcus faecalis, which is one of the most common threats observed in recurrent root canal treatment failures, of which the most problematic to treat are its biofilm-forming VRE strains. An effective response against such infections could be the use of bacteriophages (phages). Phage therapy was found to be highly effective against biofilm and multidrug-resistant bacteria and has other advantages like ease of isolation and possibilities for genetic manipulations. The potential of phage therapy in dentistry, in particular against E. faecalis biofilms in root canals, is almost unexplored. Here we review the efforts to develop phage therapy against biofilms. We also focus on the phages isolated against E. faecalis and discuss the possibility of using phages against E. faecalis biofilm in root canals. PMID:27640530

  5. Helicobacter marmotae and novel Helicobacter and Campylobacter species isolated from the livers and intestines of prairie dogs

    Science.gov (United States)

    Beisele, Maike; Shen, Zeli; Parry, Nicola; Mobley, Melissa; Taylor, Nancy S.; Buckley, Ellen; Abedin, Mohammad Z.; Dewhirst, Floyd E.

    2011-01-01

    Prairie dogs (Cynomys ludovicianus) are used to study the aetiology and prevention of gallstones because of the similarities of prairie dog and human bile gallstone composition. Epidemiological and experimental studies have suggested a connection between infection with Helicobacter species and cholesterol cholelithiasis, cholecystis and gallbladder cancer. Ten of the 34 prairie dogs in this study had positive Helicobacter species identified by PCR using Helicobacter genus-specific primers. Ten of 34 prairie dogs had positive Campylobacter species identified in the intestine by PCR with Campylobacter genus-specific primers. Six Helicobacter sp. isolates and three Campylobacter sp. isolates were identified taxonomically by 16S rRNA gene analysis. The prairie dog helicobacters fell into three clusters adjacent to Helicobacter marmotae. On the basis of 16S rRNA gene sequence analysis, three strains in two adjacent clusters were included in the species H. marmotae. Three strains were only 97.1 % similar to the sequence of H. marmotae and can be considered a novel species with the provisional designation Helicobacter sp. Prairie Dog 3. The prairie dog campylobacters formed a single novel cluster and represent a novel Campylobacter sp. with the provisional designation Campylobacter sp. Prairie Dog. They branched with Campylobacter cuniculorum at 96.3 % similarity and had the greatest sequence similarity to Campylobacter helveticus at 97.1 % similarity. Whether H. marmotae or the novel Helicobacter sp. and Campylobacter sp. identified in prairie dogs play a role in cholesterol gallstones or hepatobiliary disease requires further studies. PMID:21546560

  6. Enterite da Campylobacter upsaliensis: un paradigma della microbiologia clinica

    Directory of Open Access Journals (Sweden)

    Maria Letizia D’Annibale

    2006-12-01

    Full Text Available The Authors describe a case of acute enteritis in a adult woman with severe hepatitis C infection caused by a strain of Campylobacter upsaliensis.The clinical isolate was obtained only on blood agar because filter membrane technique was performed. In fact for this strain it was no growth on selective Campy agar, which include cephalotin too; C. upsaliensis is sensitive to this antibiotic molecule. So, the use of filter membrane on blood agar or charcoal agar at 37°C in microaerophilic atmosphere is recommended for isolation of all campylobacters responsible of human enteritis.

  7. Human Volunteers Receiving Escherichia coli Phage T4 Orally: a Safety Test of Phage Therapy

    OpenAIRE

    Bruttin, Anne; Brüssow, Harald

    2005-01-01

    Fifteen healthy adult volunteers received in their drinking water a lower Escherichia coli phage T4 dose (103 PFU/ml), a higher phage dose (105 PFU/ml), and placebo. Fecal coliphage was detected in a dose-dependent way in volunteers orally exposed to phage. All volunteers receiving the higher phage dose showed fecal phage 1 day after exposure; this prevalence was only 50% in subjects receiving the lower phage dose. No fecal phage was detectable a week after a 2-day course of oral phage applic...

  8. Intra- and inter-generic transfer of pathogenicity island-encoded virulence genes by cos phages.

    Science.gov (United States)

    Chen, John; Carpena, Nuria; Quiles-Puchalt, Nuria; Ram, Geeta; Novick, Richard P; Penadés, José R

    2015-05-01

    Bacteriophage-mediated horizontal gene transfer is one of the primary driving forces of bacterial evolution. The pac-type phages are generally thought to facilitate most of the phage-mediated gene transfer between closely related bacteria, including that of mobile genetic elements-encoded virulence genes. In this study, we report that staphylococcal cos-type phages transferred the Staphylococcus aureus pathogenicity island SaPIbov5 to non-aureus staphylococcal species and also to different genera. Our results describe the first intra- and intergeneric transfer of a pathogenicity island by a cos phage, and highlight a gene transfer mechanism that may have important implications for pathogen evolution.

  9. The effect of presence of infected neighbouring farms for the Campylobacter infection status in Danish broiler farms

    DEFF Research Database (Denmark)

    Chowdhury, Sharmin; Sandberg, Marianne; Themudo, Gonçalo E;

    2012-01-01

    Neighbourhood risk factors for Campylobacter infection in Danish broilers were evaluated. Campylobacter infection status of a flock was identified by PCR analysis of cloacal swab samples collected as a part of national surveillance program. Included into the study were, in total, 10,876 broiler...

  10. Extended multilocus sequence typing system for Campylobacter coli, C lari, C-upsaliensis, and C-helveticus

    DEFF Research Database (Denmark)

    Miller, W.G.; On, Stephen L.W.; Wang, G.L.;

    2005-01-01

    method to include three clinically relevant Campylobacter species, C. coli, C. lari, and C. upsaliensis, and a fourth Campylobacter species, C. helveticus. The C. coli and C. helveticus methods use the same seven C. jejuni loci (aspA, atpA, glnA, gltA, glyA, pgm, and tkt); however, adk and pgi were...

  11. Safety analysis of a Russian phage cocktail: From MetaGenomic analysis to oral application in healthy human subjects

    Energy Technology Data Exchange (ETDEWEB)

    McCallin, Shawna, E-mail: semccallin@yahoo.com [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland); Alam Sarker, Shafiqul, E-mail: sasarker@icddrb.org [International Centre for Diarrhoeal Diseases Research, Bangladesh (icddr,b), 68 Shaheed Tajuddin Ahmed Sharani, Mohakhali, Dhaka 1212 (Bangladesh); Barretto, Caroline, E-mail: Caroline.Barretto@rdls.nestle.com [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland); Sultana, Shamima, E-mail: shamima@icddrb.org [International Centre for Diarrhoeal Diseases Research, Bangladesh (icddr,b), 68 Shaheed Tajuddin Ahmed Sharani, Mohakhali, Dhaka 1212 (Bangladesh); Berger, Bernard, E-mail: bernard.berger@rdls.nestle.com [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland); Huq, Sayeda, E-mail: sayeeda@mail.icddrb.org [International Centre for Diarrhoeal Diseases Research, Bangladesh (icddr,b), 68 Shaheed Tajuddin Ahmed Sharani, Mohakhali, Dhaka 1212 (Bangladesh); Krause, Lutz, E-mail: ltz.krause@gmail.com [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland); Bibiloni, Rodrigo, E-mail: Rodrigo.Bibiloni@agresearch.co.nz [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland); Schmitt, Bertrand, E-mail: bertrand.schmitt@rdls.nestle.com [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland); Reuteler, Gloria, E-mail: gloria.reuteler@rdls.nestle.com [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland); Brüssow, Harald, E-mail: harald.bruessow@rdls.nestle.com [Nestlé Research Centre, Nestec Ltd, Vers-chez-les-Blanc, CH-1000 Lausanne 26 (Switzerland)

    2013-09-01

    Phage therapy has a long tradition in Eastern Europe, where preparations are comprised of complex phage cocktails whose compositions have not been described. We investigated the composition of a phage cocktail from the Russian pharmaceutical company Microgen targeting Escherichia coli/Proteus infections. Electron microscopy identified six phage types, with numerically T7-like phages dominating over T4-like phages. A metagenomic approach using taxonomical classification, reference mapping and de novo assembly identified 18 distinct phage types, including 7 genera of Podoviridae, 2 established and 2 proposed genera of Myoviridae, and 2 genera of Siphoviridae. De novo assembly yielded 7 contigs greater than 30 kb, including a 147-kb Myovirus genome and a 42-kb genome of a potentially new phage. Bioinformatic analysis did not reveal undesired genes and a small human volunteer trial did not associate adverse effects with oral phage exposure. - Highlights: • We analyzed the composition of a commercial Russian phage cocktail. • The cocktail consists of at least 10 different phage genera. • No undesired genes were detected. • No adverse effects were seen upon oral application in a small human clinical trial.

  12. Comparative characterization of the virulence gene clusters (lipooligosacharide [LOS] and capsular polysaccharide [CPS]) for Campylobacter coli, Campylobacter jejuni subsp. jejuni and related Campylobacter species

    Science.gov (United States)

    Richards, Vincent P.; Lefébure, Tristan; Pavinski Bitar, Paulina D.; Stanhope, Michael J.

    2013-01-01

    Campylobacter jejuni subsp. jejuni and Campylobacter coli are leading causes of gastroenteritis, with virulence linked to cell surface carbohydrate diversity. Although the associated gene clusters are well studied for C. jejuni subsp. jejuni, C. coli has been largely neglected. Here we provide comparative analysis of the lipooligosacharide (LOS) and capsular polysaccharide (CPS) gene clusters, using genome and cluster sequence data for 36 C. coli strains, 67 C. jejuni subsp. jejuni strains and ten additional Campylobacter species. Similar to C. jejuni subsp. jejuni, C. coli showed high LOS/CPS gene diversity, with each cluster delineated into eight gene content classes. This diversity was predominantly due to extensive gene gain/loss, with the lateral transfer of genes likely occurring both within and between species and also between the LOS and CPS. Additional mechanisms responsible for LOS/CPS diversity included phase-variable homopolymeric repeats, gene duplication/inactivation, and possibly host environment selection pressure. Analyses also showed that (i) strains of C. coli and Campylobacter upsaliensis possessed genes homologous to the sialic acid genes implicated in the neurological disorder Guillain Barré syndrome (GBS), and (ii) C. coli LOS classes were differentiated between bovine and poultry hosts, potentially aiding post infection source tracking. PMID:23279811

  13. Comparative characterization of the virulence gene clusters (lipooligosaccharide [LOS] and capsular polysaccharide [CPS]) for Campylobacter coli, Campylobacter jejuni subsp. jejuni and related Campylobacter species.

    Science.gov (United States)

    Richards, Vincent P; Lefébure, Tristan; Pavinski Bitar, Paulina D; Stanhope, Michael J

    2013-03-01

    Campylobacter jejuni subsp. jejuni and Campylobacter coli are leading causes of gastroenteritis, with virulence linked to cell surface carbohydrate diversity. Although the associated gene clusters are well studied for C. jejuni subsp. jejuni, C. coli has been largely neglected. Here we provide comparative analysis of the lipooligosaccharide (LOS) and capsular polysaccharide (CPS) gene clusters, using genome and cluster sequence data for 36 C. coli strains, 67 C. jejuni subsp. jejuni strains and ten additional Campylobacter species. Similar to C. jejuni subsp. jejuni, C. coli showed high LOS/CPS gene diversity, with each cluster delineated into eight gene content classes. This diversity was predominantly due to extensive gene gain/loss, with the lateral transfer of genes likely occurring both within and between species and also between the LOS and CPS. Additional mechanisms responsible for LOS/CPS diversity included phase-variable homopolymeric repeats, gene duplication/inactivation, and possibly host environment selection pressure. Analyses also showed that (i) strains of C. coli and Campylobacter upsaliensis possessed genes homologous to the sialic acid genes implicated in the neurological disorder Guillain-Barré syndrome (GBS), and (ii) C. coli LOS classes were differentiated between bovine and poultry hosts, potentially aiding post infection source tracking. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Spatial analysis of campylobacter infection in the Canadian province of Manitoba

    Directory of Open Access Journals (Sweden)

    Krause Dennis O

    2006-01-01

    Full Text Available Abstract Background The study describes population level variations in campylobacter incidence within the Canadian province of Manitoba, and the relationship to sociodemographic and landscape related characteristics. Using data derived from the Manitoba Health Public Health Branch communicable disease surveillance database, the study applied a number of spatial and ecological techniques to visualize, explore and model campylobacter incidence for the years 1996 to 2004. Analytical techniques used in the study included spatial smoothing, the spatial scan statistic, the Gini coefficient, and Poisson regression analysis. Results The study demonstrated marked and statistically significant geographic variability in the rates of campylobacter incidence in Manitoba.. The incidence of campylobacter was observed to be significantly higher in populations living in rural and agricultural areas of the province, with the highest rates occurring in populations living in proximity to high densities of farm animals (cows, pigs, chickens. The study also observed that the age specific pattern of campylobacter incidence in rural Manitoba was very different than the urban pattern, with the incidence rate in the 0–4 year age group seven times higher in rural Manitoba than in the City of Winnipeg. Conclusion The study demonstrates the value of a deploying a diverse set of spatial techniques to better understand the dynamics of an enteric disease such as campylobacter infection. The study concludes that there may be three distinct mechanisms for the transmission of campylobacter in Manitoba which are operating simultaneously. These include broad population exposure to a centralized food system endemically infected with the campylobacter organism, exposure to local level factors such as farm animals or contaminated water, and exposure to campylobacter infection through foreign travel.

  15. Genomic Evidence for the Emergence and Evolution of Pathogenicity and Niche Preferences in the Genus Campylobacter

    Science.gov (United States)

    Iraola, Gregorio; Pérez, Ruben; Naya, Hugo; Paolicchi, Fernando; Pastor, Eugenia; Valenzuela, Sebastián; Calleros, Lucía; Velilla, Alejandra; Hernández, Martín; Morsella, Claudia

    2014-01-01

    The genus Campylobacter includes some of the most relevant pathogens for human and animal health; the continuous effort in their characterization has also revealed new species putatively involved in different kind of infections. Nowadays, the available genomic data for the genus comprise a wide variety of species with different pathogenic potential and niche preferences. In this work, we contribute to enlarge this available information presenting the first genome for the species Campylobacter sputorum bv. sputorum and use this and the already sequenced organisms to analyze the emergence and evolution of pathogenicity and niche preferences among Campylobacter species. We found that campylobacters can be unequivocally distinguished in established and putative pathogens depending on their repertory of virulence genes, which have been horizontally acquired from other bacteria because the nonpathogenic Campylobacter ancestor emerged, and posteriorly interchanged between some members of the genus. Additionally, we demonstrated the role of both horizontal gene transfers and diversifying evolution in niche preferences, being able to distinguish genetic features associated to the tropism for oral, genital, and gastrointestinal tissues. In particular, we highlight the role of nonsynonymous evolution of disulphide bond proteins, the invasion antigen B (CiaB), and other secreted proteins in the determination of niche preferences. Our results arise from assessing the previously unmet goal of considering the whole available Campylobacter diversity for genome comparisons, unveiling notorious genetic features that could explain particular phenotypes and set the basis for future research in Campylobacter biology. PMID:25193310

  16. Genomic evidence for the emergence and evolution of pathogenicity and niche preferences in the genus Campylobacter.

    Science.gov (United States)

    Iraola, Gregorio; Pérez, Ruben; Naya, Hugo; Paolicchi, Fernando; Pastor, Eugenia; Valenzuela, Sebastián; Calleros, Lucía; Velilla, Alejandra; Hernández, Martín; Morsella, Claudia

    2014-09-04

    The genus Campylobacter includes some of the most relevant pathogens for human and animal health; the continuous effort in their characterization has also revealed new species putatively involved in different kind of infections. Nowadays, the available genomic data for the genus comprise a wide variety of species with different pathogenic potential and niche preferences. In this work, we contribute to enlarge this available information presenting the first genome for the species Campylobacter sputorum bv. sputorum and use this and the already sequenced organisms to analyze the emergence and evolution of pathogenicity and niche preferences among Campylobacter species. We found that campylobacters can be unequivocally distinguished in established and putative pathogens depending on their repertory of virulence genes, which have been horizontally acquired from other bacteria because the nonpathogenic Campylobacter ancestor emerged, and posteriorly interchanged between some members of the genus. Additionally, we demonstrated the role of both horizontal gene transfers and diversifying evolution in niche preferences, being able to distinguish genetic features associated to the tropism for oral, genital, and gastrointestinal tissues. In particular, we highlight the role of nonsynonymous evolution of disulphide bond proteins, the invasion antigen B (CiaB), and other secreted proteins in the determination of niche preferences. Our results arise from assessing the previously unmet goal of considering the whole available Campylobacter diversity for genome comparisons, unveiling notorious genetic features that could explain particular phenotypes and set the basis for future research in Campylobacter biology.

  17. Phage morphology recapitulates phylogeny: the comparative genomics of a new group of myoviruses.

    Directory of Open Access Journals (Sweden)

    André M Comeau

    Full Text Available Among dsDNA tailed bacteriophages (Caudovirales, members of the Myoviridae family have the most sophisticated virion design that includes a complex contractile tail structure. The Myoviridae generally have larger genomes than the other phage families. Relatively few "dwarf" myoviruses, those with a genome size of less than 50 kb such as those of the Mu group, have been analyzed in extenso. Here we report on the genome sequencing and morphological characterization of a new group of such phages that infect a diverse range of Proteobacteria, namely Aeromonas salmonicida phage 56, Vibrio cholerae phages 138 and CP-T1, Bdellovibrio phage φ1422, and Pectobacterium carotovorum phage ZF40. This group of dwarf myoviruses shares an identical virion morphology, characterized by usually short contractile tails, and have genome sizes of approximately 45 kb. Although their genome sequences are variable in their lysogeny, replication, and host adaption modules, presumably reflecting differing lifestyles and hosts, their structural and morphogenesis modules have been evolutionarily constrained by their virion morphology. Comparative genomic analysis reveals that these phages, along with related prophage genomes, form a new coherent group within the Myoviridae. The results presented in this communication support the hypothesis that the diversity of phages may be more structured than generally believed and that the innumerable phages in the biosphere all belong to discrete lineages or families.

  18. Complete Genome Sequence of Vibrio anguillarum Phage CHOED Successfully Used for Phage Therapy in Aquaculture

    OpenAIRE

    Romero, Jaime; Higuera, Gastón; Gajardo,Felipe; Castillo, Daniel; Middleboe, Mathias; García, Katherine; Ramírez, Carolina; Espejo, Romilio T.

    2014-01-01

    Vibrio anguillarum phage CHOED was isolated from Chilean mussels. It is a virulent phage showing effective inhibition of V. anguillarum. CHOED has potential in phage therapy, because it can protect fish from vibriosis in fish farms. Here, we announce the completely sequenced genome of V. anguillarum phage CHOED.

  19. Survey of Campylobacter spp. in owned and unowned dogs and cats in Northern Italy.

    Science.gov (United States)

    Giacomelli, M; Follador, N; Coppola, L M; Martini, M; Piccirillo, A

    2015-06-01

    Campylobacteriosis is among the most common bacterial causes of human gastroenteritis worldwide and pet ownership has been identified as a risk factor for Campylobacter infection in humans. Since canine and feline prevalence data are scarce in Italy, the present study was carried out to assess the prevalence, species distribution and risk factors for Campylobacter infection in dogs and cats under different husbandry conditions. Rectal swabs were collected from 171 dogs (household pets, n = 100; shelter-housed dogs, n = 50; dogs from breeding kennels, n = 21) and 102 cats (household pets, n = 52; shelter-housed cats, n = 21; free-roaming cats n = 29) in Northern Italy. Campylobacter was isolated from 17% (n = 29) of dogs and 14.7% (n = 15) of cats. C. jejuni was the most common isolate in both species (Campylobacter spp.-positive dogs, 55.2%; Campylobacter spp.-positive cats, 53.3%), followed by C. upsaliensis (Campylobacter spp.-positive dogs, 27.6%; Campylobacter spp.-positive cats, 40%). Other Campylobacter species were rarely detected, but included C. hyointestinalis subsp. hyointestinalis, C. lari and C. coli in dogs and C. coli and C. helveticus in cats. Among considered variables (sex, age, origin, diarrhoea, season of sampling), origin was identified as a risk factor for dogs, with shelter-housed dogs at higher risk than household dogs (odds ratio, 2.84; 95% CI 1.17, 6.92; P = 0.021). The results of this study, particularly the high prevalence of C. jejuni in Campylobacter-positive animals, demonstrated that household and stray dogs and cats in Northern Italy might pose a zoonotic risk for humans. Moreover, biosecurity measures should be improved in dog shelters. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Information Phage Therapy Research Should Report

    National Research Council Canada - National Science Library

    Stephen T Abedon

    2017-01-01

    .... Clinically these applications are described as phage or bacteriophage therapy. Alternatively, and especially in the treatment of environments, this practice instead may be described as a phage-mediated biocontrol of bacteria...

  1. Campylobacter bacteremia: A rare and under-reported event?

    NARCIS (Netherlands)

    Louwen, R.; Baarlen, van P.; Vliet, van A.H.M.; Belkum, van A.; Hays, J.P.; Endtz, H.P.

    2012-01-01

    Bacteria belonging to the species Campylobacter are the most common cause of bacterial diarrhoea in humans. The clinical phenotype associated with Campylobacter infections ranges from asymptomatic conditions to severe colitis and bacteremia. In susceptible patients, Campylobacter infections are

  2. Campylobacter bacteremia: A rare and under-reported event?

    NARCIS (Netherlands)

    Louwen, R.; Baarlen, van P.; Vliet, van A.H.M.; Belkum, van A.; Hays, J.P.; Endtz, H.P.

    2012-01-01

    Bacteria belonging to the species Campylobacter are the most common cause of bacterial diarrhoea in humans. The clinical phenotype associated with Campylobacter infections ranges from asymptomatic conditions to severe colitis and bacteremia. In susceptible patients, Campylobacter infections are asso

  3. Novel Campylobacter isolation method using hydrophobic grid membrane filter and semisolid medium.

    Science.gov (United States)

    Valdivieso-Garcia, Alfonso; Harris, Kathleen; Riche, Edward; Campbell, Stephanie; Jarvie, Anne; Popa, Maria; Deckert, Anne; Reid-Smith, Richard; Rahn, Kris

    2007-02-01

    Culture procedures for isolation of thermophilic campylobacters from food matrices are complex, labor intensive, and time-consuming. Most available methods include the use of antibiotics as selective agents to prevent the growth of competing microflora. A simple procedure for isolation of thermophilic campylobacters after enrichment in Rosef's enrichment broth was developed using a hydrophobic grid membrane filter (HGMF) on semisolid medium (SSM). SSM contains no antibiotics, and the HGMF physically separates Campylobacter from the enrichment broth, allowing isolation based on differential motility. The HGMF-SSM method was compared to the Agriculture and Agri-Food Canada Food Safety Procedures Manual (FSPM-10) method (Isolation of Thermophilic Campylobacters from Fresh Pork, Beef Veal, Poultry and Ready-to-Eat Meat Products), which includes the use of selective antibiotics. During the initial study, after enrichment the HGMF-SSM method yielded pure cultures of campylobacters after 16 to 18 h (overnight) compared with 48 h for the FSPM-10 method. Ninety-four turkey samples collected at local retail stores and 38 frozen pig fecal samples were processed by both methods. Thirty-five samples (26.5%) were positive by the HGMF-SSM method; 24 (18.2%) of these positive samples contained Campylobacter jejuni and 11 (8.3%) contained Campylobacter coli. With the FSPM-10 method, 25 samples (18.9%) were positive: 21 (15.9%) with C. jejuni and 4 (3%) with C. coli. For a subsequent field study, only the HGMF-SSM method was used to isolate Campylobacter from 1,200 chicken samples and 454 turkey samples sold at retail. Analysis of five subisolates from various samples indicated that only one type of Campylobacter was recovered by the HGMF-SSM method, as ascertained by MICs for 10 antimicrobials, sequencing of the short variable region of the flaA gene, and fingerprinting based on amplified fragment length polymorphism. The absence of antibiotics in the SSM may explain the higher

  4. Antimicrobial resistance of thermophilic Campylobacter

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Engberg, J.

    2001-01-01

    Campylobacter has become the leading cause of zoonotic enteric infections in developed and developing countries world-wide. Antimicrobial resistance has emerged among Campylobacter mainly as a consequence of the use of antimicrobial agents in food animal production. Resistance to drugs of choice...... for the treatment of infections, macrolides and fluoroquinolones has emerged as a clinical problem and interventions to reduce this are recommended. Resistance to fluoroquinolones and macrolides is mediated by chromosomal mutations. Resistance to other relevant antimicrobial agents, mediated by acquired resistance...

  5. The ability of flagellum-specific Proteus vulgaris bacteriophage PV22 to interact with Campylobacter jejuni flagella in culture

    Directory of Open Access Journals (Sweden)

    Stern NJ

    2006-06-01

    Full Text Available Abstract Background There has been a recent resurgent interest in bacteriophage biology. Research was initiated to examine Campylobacter jejuni-specific bacteriophage in the Russian Federation to develop alternative control measures for this pathogen. Results A C. jejuni flagellum-specific phage PV22 from Proteus vulgaris was identified in sewage drainage. This phage interacted with C. jejuni by attachment to flagella followed by translocation of the phage to the polar region of the bacterium up to the point of DNA injection. Electron microscopic examination revealed adsorption of PV22 on C. jejuni flagella after a five minute incubation of the phage and bacteria. A different phenomenon was observed after incubating the mix under the same conditions, but for twenty minutes or longer. Phage accumulated primarily on the surface of cells at sites where flagella originated. Interestingly, PV22 did not inject DNA into C. jejuni and PV22 did not produce lytic plaques on medium containing C. jejuni cells. The constant of velocity for PV22 adsorption on cells was 7 × 10-9 ml/min. Conclusion It was demonstrated that a bacteriophage that productively infects P. vulgaris was able to bind C. jejuni and by a spot test that the growth of C. jejuni was reduced relative to control bacteria in the region of phage application. There may be two interesting applications of this effect. First, it may be possible to test phage PV22 as an antimicrobial agent to decrease C. jejuni colonization of the chicken intestine. Second, the phage could potentially be utilized for investigating biogenesis of C. jejuni flagella.

  6. Phage Therapy: Eco-Physiological Pharmacology

    OpenAIRE

    Abedon, Stephen T.

    2014-01-01

    Bacterial virus use as antibacterial agents, in the guise of what is commonly known as phage therapy, is an inherently physiological, ecological, and also pharmacological process. Physiologically we can consider metabolic properties of phage infections of bacteria and variation in those properties as a function of preexisting bacterial states. In addition, there are patient responses to pathogenesis, patient responses to phage infections of pathogens, and also patient responses to phage virio...

  7. Phage as a Genetically Modifiable Supramacromolecule in Chemistry, Materials and Medicine.

    Science.gov (United States)

    Cao, Binrui; Yang, Mingying; Mao, Chuanbin

    2016-06-21

    differentiation; (4) discovered that phage could induce angiogenesis and osteogenesis for MSC-based vascularized bone regeneration; (5) identified novel breast cancer cell-targeting and MSC-targeting peptides and used them to significantly improve the efficiency of targeted cancer therapy and MSC-based gene delivery, respectively; (6) employed engineered phage as a probe to achieve ultrasensitive detection of biomarkers from serum of human patients for disease diagnosis; and (7) constructed centimeter-scale 3D multilayered phage assemblies with the potential application as scaffolds for bone regeneration and functional device fabrication. Our findings demonstrated that phage is indeed a very powerful supramacromolecule suitable for not only developing novel nanostructures and biomaterials but also advancing important fields in biomedicine, including molecular targeting, cancer diagnosis and treatment, drug and gene delivery, stem cell fate direction, and tissue regeneration. Our successes in exploiting phage in chemistry, materials, and medicine suggest that phage itself is nontoxic at the cell level and can be safely used for detecting biomarkers in vitro. Moreover, although we have demonstrated successful in vivo tissue regeneration induced by phage, we believe future studies are needed to evaluate the in vivo biodistribution and potential risks of the phage-based biomaterials.

  8. The complete genome sequence and analysis of the human pathogen Campylobacter lari

    DEFF Research Database (Denmark)

    Miller, WG; Wang, G; Binnewies, Tim Terence

    2008-01-01

    Campylobacter lari is a member of the epsilon subdivision of the Proteobacteria and is part of the thermotolerant Campylobacter group, a clade that includes the human pathogen C. jejuni. Here we present the complete genome sequence of the human clinical isolate, C. lari RM2100. The genome of strain...... RM2100 is approximately 1.53 Mb and includes the 46 kb megaplasmid pCL2100. Also present within the strain RM2100 genome is a 36 kb putative prophage, termed CLIE1, which is similar to CJIE4, a putative prophage present within the C. jejuni RM1221 genome. Nearly all (90%) of the gene content...... in strain RM2100 is similar to genes present in the genomes of other characterized thermotolerant campylobacters. However, several genes involved in amino acid biosynthesis and energy metabolism, identified previously in other Campylobacter genomes, are absent from the C. lari RM2100 genome. Therefore, C...

  9. Phage therapy--constraints and possibilities.

    Science.gov (United States)

    Nilsson, Anders S

    2014-05-01

    The rise of antibiotic-resistant bacterial strains, causing intractable infections, has resulted in an increased interest in phage therapy. Phage therapy preceded antibiotic treatment against bacterial infections and involves the use of bacteriophages, bacterial viruses, to fight bacteria. Virulent phages are abundant and have proven to be very effective in vitro, where they in most cases lyse any bacteria within the hour. Clinical trials on animals and humans show promising results but also that the treatments are not completely effective. This is partly due to the studies being carried out with few phages, and with limited experimental groups, but also the fact that phage therapy has limitations in vivo. Phages are large compared with small antibiotic molecules, and each phage can only infect one or a few bacterial strains. A very large number of different phages are needed to treat infections as these are caused by genetically different strains of bacteria. Phages are effective only if enough of them can reach the bacteria and increase in number in situ. Taken together, this entails high demands on resources for the construction of phage libraries and the testing of individual phages. The effectiveness and host range must be characterized, and immunological risks must be assessed for every single phage.

  10. Synthetic Phage for Tissue Regeneration

    Directory of Open Access Journals (Sweden)

    So Young Yoo

    2014-01-01

    Full Text Available Controlling structural organization and signaling motif display is of great importance to design the functional tissue regenerating materials. Synthetic phage, genetically engineered M13 bacteriophage has been recently introduced as novel tissue regeneration materials to display a high density of cell-signaling peptides on their major coat proteins for tissue regeneration purposes. Structural advantages of their long-rod shape and monodispersity can be taken together to construct nanofibrous scaffolds which support cell proliferation and differentiation as well as direct orientation of their growth in two or three dimensions. This review demonstrated how functional synthetic phage is designed and subsequently utilized for tissue regeneration that offers potential cell therapy.

  11. Campylobacter hyoilei Alderton et al. 1995 and Campylobacter coli Veron and Chatelain 1973 are subjective synonyms

    DEFF Research Database (Denmark)

    Vandamme, P.; VanDoorn, L.J.; AlRashid, S.T.

    1997-01-01

    The taxonomic affiliation of Campylobacter hyoilei was reevaluated by examining a variety of phenotypic and genotypic criteria. Whole cell protein electrophoresis and a comparison of 66 phenotypic characters revealed that reference strains of C. hyoilei were indistinguishable from Campylobacter c...

  12. Sequence analysis of the phage 21 genes for prohead assembly and head completion.

    Science.gov (United States)

    Smith, M P; Feiss, M

    1993-04-15

    Phage 21 is a temperate lambdoid coliphage, and its head-encoding genes, as well as those of phage lambda, are descended from a common ancestral phage. The head protein-encoding genes of phage 21 have been sequenced, confirming earlier genetic studies indicating that the head-encoding genes of 21 and lambda are analogous in location, size, and function. The phage 21 head-encoding genes identified (and their lambda analogues) include: 3(W), 4(B), 5(C), 6(Nu3), shp (D), 7(E), and 8(FII), respectively. An open reading frame, orf1, is analogous in position and shares some sequence identity with FI, a phage lambda gene involved in DNA packaging. The phage 21 major head protein, gp7, is predicted to have strong sequence identity (65%) with the lambda major capsid protein, gpE, including amino acids known to be important for capsid form determination. The nested genes 5/6 of phage 21 and C/Nu3 of lambda differ by several rearrangements including deletions and a triplication. The possibility that lambda genes C/Nu3 evolved from ancestal nested genes containing a triplication is discussed.

  13. Occurrence of Campylobacter jejuni and Campylobacter coli and their biotypes in beef and dairy cattle from the south of Chile

    OpenAIRE

    Heriberto Fernández; Marianne Hitschfeld

    2009-01-01

    The prevalence of Campylobacter jejuni and Campylobacter coli and their biotypes in beef and dairy cattle from the South of Chile was established. Campylobacter were statistically more prevalent among beef cattle (35.9%) than among dairy cattle (21.3%), being C. jejuni the species most frequently isolated.Foi estabelecida a prevalência de Campylobacter jejuni e Campylobacter coli e seus biotipos, em bovinos de corte e de leite do sul do Chile. Campylobacter foi estatisticamente mais prevalent...

  14. Distribution and Polymorphism of the Flagellin Genes from Isolates of Campylobacter coli and Campylobacter jejuni

    Science.gov (United States)

    1993-05-01

    American Society for Microbioloc% Distribution and Polymorphism of the Flagellin Genes from Isolates of Campylobacter coli and Campylobacter jejuni RICHARD...in Campylobacter jejuni . serogroups both the flaA and flaB genes are extremely Mol. M;crobiol. 5:1151-1158. z homologous. Within most LIO heat-labile...irllwn hungatei. J1. Bacteriol. 123:-28 proteins of Campylobacter jejuni 81116. Infect. Immun. 59: 42. Thomashow, L S., and S. C. Rittenberg. 198

  15. Specific pathogen-free pig herds also free from Campylobacter?

    Science.gov (United States)

    Kolstoe, E M; Iversen, T; Østensvik, Ø; Abdelghani, A; Secic, I; Nesbakken, T

    2015-03-01

    As Specific Pathogen-Free (SPF) pig herds are designed and managed to prevent specific pig diseases, it might be feasible to expand the list of micro-organisms also including zoonotic pathogens such as Campylobacter coli as this agent has its origin in pigs. In a previous survey, 15 of 16 of SPF herds were found free from human pathogenic Yersinia enterocolitica. Accordingly, three nucleus and seven multiplying herds were surveyed for Campylobacter to investigate whether the Norwegian SPF pig pyramid also might be free from this agent. In conclusion, the intervention of Campylobacter at the herd level might be possible as four of 10 SPF herds tested negative in two sets of samples from both autumn 2008 and summer/early autumn 2010. The four negative herds were all located in remote areas several kilometres away from conventional pig farming while the positive SPF farms were all situated in neighbourhoods with conventional pig production. It seems more difficult to control Campylobacter than some specific animal disease agents and another significant zoonotic agent, Y. enterocolitica, in pig herds. © 2014 Blackwell Verlag GmbH.

  16. Role of emerging Campylobacter species in inflammatory bowel diseases.

    Science.gov (United States)

    Kaakoush, Nadeem O; Mitchell, Hazel M; Man, Si Ming

    2014-11-01

    The gut microbiota is a central player in the etiology of inflammatory bowel diseases. As such, there is intense scientific interest in elucidating the specific group/s of bacteria responsible for driving barrier damage and perpetuating the chronic inflammation that results in disease. Because of their ability to colonize close to the surface of the host intestinal epithelium, mucosa-associated bacteria are considered key players in the initiation and development of both Crohn's disease and ulcerative colitis. The leading bacterial candidates include adherent and invasive Escherichia coli, Helicobacter, Fusobacteria, Mycobacteria, and Campylobacter species. Of these, a member of the Campylobacter genus, Campylobacter concisus, has recently emerged as a putative player in the pathogenesis of inflammatory bowel diseases. Current research indicates that this bacterium possesses extraordinarily diverse pathogenic capacities as well as unique genetic and functional signatures that are defined by their ability to adhere to and invade host cells, secrete toxins, and the presence of a virulence-associated restriction-modification system. These characteristics enable the potential classification of C. concisus into distinct pathotypes, which we have named adherent and invasive C. concisus and adherent and toxinogenic C. concisus. In this review, we evaluate evidence for the role of emerging Campylobacter species in the pathogenesis of inflammatory bowel diseases.

  17. Nongenetic individuality in the host-phage interaction.

    Directory of Open Access Journals (Sweden)

    Sivan Pearl

    2008-05-01

    Full Text Available Isogenic bacteria can exhibit a range of phenotypes, even in homogeneous environmental conditions. Such nongenetic individuality has been observed in a wide range of biological processes, including differentiation and stress response. A striking example is the heterogeneous response of bacteria to antibiotics, whereby a small fraction of drug-sensitive bacteria can persist under extensive antibiotic treatments. We have previously shown that persistent bacteria enter a phenotypic state, identified by slow growth or dormancy, which protects them from the lethal action of antibiotics. Here, we studied the effect of persistence on the interaction between Escherichia coli and phage lambda. We used long-term time-lapse microscopy to follow the expression of green fluorescent protein (GFP under the phage lytic promoter, as well as cellular fate, in single infected bacteria. Intriguingly, we found that, whereas persistent bacteria are protected from prophage induction, they are not protected from lytic infection. Quantitative analysis of gene expression reveals that the expression of lytic genes is suppressed in persistent bacteria. However, when persistent bacteria switch to normal growth, the infecting phage resumes the process of gene expression, ultimately causing cell lysis. Using mathematical models for these two host-phage interactions, we found that the bacteria's nongenetic individuality can significantly affect the population dynamics, and might be relevant for understanding the coevolution of bacterial hosts and phages.

  18. Inflammasome activation by Campylobacter jejuni

    NARCIS (Netherlands)

    Bouwman, Lieneke I; de Zoete, Marcel R; Bleumink-Pluym, Nancy M C; Flavell, Richard A; van Putten, Jos P M

    2014-01-01

    The Gram-negative pathogen Campylobacter jejuni is the most common cause of bacterial foodborne disease worldwide. The mechanisms that lead to bacterial invasion of eukaryotic cells and massive intestinal inflammation are still unknown. In this study, we report that C. jejuni infection of mouse macr

  19. Lectin typing of Campylobacter concisus

    DEFF Research Database (Denmark)

    Aabenhus, Rune Munck; Hynes, Sean O; Permin, Henrik

    2002-01-01

    A total of 44 clinical isolates and the type strain of the putative pathogen Campylobacter concisus were grouped based on their reactions with plant lectins. The optimized lectin typing system used C. concisus strains proteolytically pretreated and subsequently typed by using a panel of four...

  20. Protozoa: a novel Campylobacter reservoir?

    Science.gov (United States)

    In previous in vitro studies we found that Campylobacter jejuni remained viable for longer periods of time when they were cultivated in the presence of Tetrahymena pyriformis (ciliate) and Acanthamoeba castellanii (amoeba) than when they were in an independent planktonic state. Increased survival t...

  1. Evaluation of 11 PCR assays for species-level identification of Campylobacter jejuni and Campylobacter coli

    DEFF Research Database (Denmark)

    On, Stephen L.W.; Jordan, Penelope J.

    2003-01-01

    We examined the sensitivity and specificity of 11 PCR assays described for the species identification of Campylobacter jejuni and Campylobacter coli by using 111 type, reference, and field strains of C. jejuni, C. coli, and Campylobacter lari. For six assays, an additional 21 type strains...

  2. Serotypes and typability of Campylobacter jejuni and Campylobacter coli isolated from poultry products

    DEFF Research Database (Denmark)

    Nielsen, Eva Møller; Nielsen, Niels Ladefoged

    1999-01-01

    Campylobacter infection is one of the most common bacterial enteric pathogens. Campylobacter jejuni and Campylobacter coli infections are mostly food- and waterborne and especially poultry is often assumed to be an important source. The heat-stable serotyping system (the 'Penner' scheme) was used...

  3. Quantitative risk assessment of human campylobacteriosis associated with thermophilic Campylobacter species in chickens

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Nielsen, N. L.; Sommer, Helle Mølgaard

    2003-01-01

    A quantitative risk assessment comprising the elements hazard identification, hazard characterization, exposure assessment, and risk characterization has been prepared to assess the effect of different mitigation strategies on the number of human cases in Denmark associated with thermophilic...... covers the transfer of Campylobacter during food handling in private kitchens. The age and sex of consumers were included in this module to introduce variable hygiene levels during food preparation and variable sizes and compositions of meals. Finally, the outcome of the exposure assessment modules...... Campylobacter spp. in chickens. To estimate the human exposure to Campylobacter from a chicken meal and the number of human cases associated with this exposure, a mathematical risk model was developed. The model details the spread and transfer of Campylobacter in chickens from slaughter to consumption...

  4. Effect of climate and farm environment on Campylobacter spp. colonisation in Norwegian broiler flocks

    DEFF Research Database (Denmark)

    Jonsson, Malin E.; Chriél, Mariann; Norström, Madelaine

    2012-01-01

    of Campylobacter spp. in Norwegian broiler flocks and factors related to the climate and the farm environment. Data from 18,488 broiler flocks from 623 different farms during 2002–2007 were included in the study. A logistic regression analysis was conducted where Campylobacter spp. status of a broiler flock...... at the time of slaughter was defined as the dependent variable and farm was modelled as a random effect. The following factors were found to increase the probability for a broiler flock to test positive for Campylobacter spp.: daily mean temperature above 6°C during the rearing period, private water supply......, presence of other livestock farms within a distance of 2km, presence of other broiler farms within a distance of 4km with flocks positive for Campylobacter spp. within 30 days prior to slaughter, heavy rainfall 11–30 days prior to slaughter, region and year. Daily mean temperature below 0°C reduced...

  5. Detection of Campylobacter jejuni and Campylobacter coli in environmental waters by PCR enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Sails, A D; Bolton, F J; Fox, A J; Wareing, D R A; Greenway, D L A

    2002-03-01

    A PCR enzyme-linked immunosorbent assay (ELISA) assay was applied to the detection of Campylobacter jejuni and Campylobacter coli in environmental water samples after enrichment culture. Bacterial cells were concentrated from 69 environmental water samples by using filtration, and the filtrates were cultured in Campylobacter blood-free broth. After enrichment culture, DNA was extracted from the samples by using a rapid-boiling method, and the DNA extracts were used as a template in a PCR ELISA assay. A total of 51 samples were positive by either PCR ELISA or culture; of these, 43 were found to be positive by PCR ELISA and 43 were found to be positive by culture. Overall, including positive and negative results, 59 samples were concordant in both methods. Several samples were positive in the PCR ELISA assay but were culture negative; therefore, this assay may be able to detect sublethally damaged or viable nonculturable forms of campylobacters. The method is rapid and sensitive, and it significantly reduces the time needed for the detection of these important pathogens by 2 to 3 days.

  6. Differential Distribution of Type II CRISPR-Cas Systems in Agricultural and Nonagricultural Campylobacter coli and Campylobacter jejuni Isolates Correlates with Lack of Shared Environments.

    Science.gov (United States)

    Pearson, Bruce M; Louwen, Rogier; van Baarlen, Peter; van Vliet, Arnoud H M

    2015-09-02

    CRISPR (clustered regularly interspaced palindromic repeats)-Cas (CRISPR-associated) systems are sequence-specific adaptive defenses against phages and plasmids which are widespread in prokaryotes. Here we have studied whether phylogenetic relatedness or sharing of environmental niches affects the distribution and dissemination of Type II CRISPR-Cas systems, first in 132 bacterial genomes from 15 phylogenetic classes, ranging from Proteobacteria to Actinobacteria. There was clustering of distinct Type II CRISPR-Cas systems in phylogenetically distinct genera with varying G+C%, which share environmental niches. The distribution of CRISPR-Cas within a genus was studied using a large collection of genome sequences of the closely related Campylobacter species Campylobacter jejuni (N = 3,746) and Campylobacter coli (N = 486). The Cas gene cas9 and CRISPR-repeat are almost universally present in C. jejuni genomes (98.0% positive) but relatively rare in C. coli genomes (9.6% positive). Campylobacter jejuni and agricultural C. coli isolates share the C. jejuni CRISPR-Cas system, which is closely related to, but distinct from the C. coli CRISPR-Cas system found in C. coli isolates from nonagricultural sources. Analysis of the genomic position of CRISPR-Cas insertion suggests that the C. jejuni-type CRISPR-Cas has been transferred to agricultural C. coli. Conversely, the absence of the C. coli-type CRISPR-Cas in agricultural C. coli isolates may be due to these isolates not sharing the same environmental niche, and may be affected by farm hygiene and biosecurity practices in the agricultural sector. Finally, many CRISPR spacer alleles were linked with specific multilocus sequence types, suggesting that these can assist molecular epidemiology applications for C. jejuni and C. coli.

  7. Biochemical functionalization of peptide nanotubes with phage displayed peptides

    Science.gov (United States)

    Swaminathan, Swathi; Cui, Yue

    2016-09-01

    The development of a general approach for the biochemical functionalization of peptide nanotubes (PNTs) could open up existing opportunities in both fundamental studies as well as a variety of applications. PNTs are spontaneously assembled organic nanostructures made from peptides. Phage display has emerged as a powerful approach for identifying selective peptide binding motifs. Here, we demonstrate for the first time the biochemical functionalization of PNTs via peptides identified from a phage display peptide library. The phage-displayed peptides are shown to recognize PNTs. These advances further allow for the development of bifunctional peptides for the capture of bacteria and the self-assembly of silver particles onto PNTs. We anticipate that these results could provide significant opportunities for using PNTs in both fundamental studies and practical applications, including sensors and biosensors nanoelectronics, energy storage devices, drug delivery, and tissue engineering.

  8. Phage-Host Interactions in Flavobacterium psychrophilum and the Potential for Phage Therapy in Aquaculture

    DEFF Research Database (Denmark)

    Christiansen, Rói Hammershaimb

    , the increasing problem with antibiotic resistance has led to increased attention to the use of phages for controlling F. psychrophilum infections in aquaculture. In a synopsis and four scientific papers, this PhD project studies the potential and optimizes the use of phage therapy for treatment and prevention...... of F. psychrophilum infections in rainbow trout fry. In the first paper, studies of the controlling effect of different phages infecting F. psychrophilum in liquid cultures showed that a high initial phage concentration was crucial for fast and effective bacterial lysis in the cultures and sensitive...... cells could be maintained at a low level throughout the rest of the experiment. Surprisingly, no difference was observed between infection with single phages or phage cocktails. At the end of incubation phage-sensitive strains dominated in the cultures with low initial phage concentrations and phage...

  9. Prevalence and Antimicrobial Resistance of Campylobacter Isolated from Dressed Beef Carcasses and Raw Milk in Tanzania.

    Science.gov (United States)

    Kashoma, Isaac P; Kassem, Issmat I; John, Julius; Kessy, Beda M; Gebreyes, Wondwossen; Kazwala, Rudovick R; Rajashekara, Gireesh

    2016-01-01

    Campylobacter species are commonly transmitted to humans through consumption of contaminated foods such as milk and meat. The aim of this study was to investigate the prevalence, antimicrobial resistance, and genetic determinants of resistance of Campylobacter isolated from raw milk and beef carcasses in Tanzania. The antimicrobial resistance genes tested included blaOXA-61 (ampicillin), aph-3-1 (aminoglycoside), tet(O) (tetracycline), and cmeB (multi-drug efflux pump). The prevalence of Campylobacter was 9.5% in beef carcasses and 13.4% in raw milk, respectively. Using multiplex-polymerase chain reaction (PCR), we identified 58.1% of the isolates as Campylobacter jejuni, 30.7% as Campylobacter coli, and 9.7% as other Campylobacter spp. One isolate (1.6%) was positive for both C. jejuni and C. coli specific PCR. Antimicrobial susceptibility testing using the disk diffusion assay and the broth microdilution method showed resistance to: ampicillin (63% and 94.1%), ciprofloxacin (9.3% and 11.8%), erythromycin (53.7% and 70.6%), gentamicin (0% and 15.7%), streptomycin (35.2% and 84.3%), and tetracycline (18.5% and 17.7%), respectively. Resistance to azithromycin (42.6%), nalidixic acid (64.8%), and chloramphenicol (13%) was determined using the disk diffusion assay only, while resistance to tylosin (90.2%) was quantified using the broth microdilution method. The blaOXA-61 (52.6% and 28.1%), cmeB (26.3% and 31.3%), tet(O) (26.3% and 31.3%), and aph-3-1 (5.3% and 3.0%) were detected in C. coli and C. jejuni. These findings highlight the extent of antimicrobial resistance in Campylobacter occurring in important foods in Tanzania. The potential risks to consumers emphasize the need for adequate control approaches, including the prudent use of antimicrobials to minimize the spread of antimicrobial-resistant Campylobacter.

  10. Occurrence of fastidious Campylobacter spp. in fresh meat and poultry using an adapted cultural protocol.

    Science.gov (United States)

    Lynch, Orla A; Cagney, Claire; McDowell, David A; Duffy, Geraldine

    2011-11-01

    This study used an adapted cultural protocol for the recovery of fastidious species of Campylobacter, to gain a more accurate understanding of the diversity of Campylobacter populations in fresh meats. Chicken (n=185), pork (n=179) and beef (n=186) were collected from supermarkets and butchers throughout the Republic of Ireland. Samples were enriched in Campylobacter enrichment broth for 24h under an atmosphere of 2.5% O(2), 7% H(2), 10% CO(2), and 80.5% N(2). The enriched samples were then filtered onto non-selective Anaerobe Basal Agar supplemented with lysed horse blood using mixed ester filter membranes. Isolates were identified by both genus and species-specific PCR assays and biochemical testing. The incidence of campylobacters on beef (36%) was significantly higher than on pork (22%) or chicken (16%), and far exceeds previously reported prevalence levels. The method was successful in recovering 7 species of Campylobacter, including the fastidious spp. C. concisus and C. mucosalis, from chicken meat, and 10 species, including C. concisus, C. curvus, C. mucosalis, C. sputorum, and C. upsaliensis, from minced beef. The isolation of C. concisus and C. upsaliensis from meat in this study is of particular significance, due to their emerging clinical relevance. The results of this study confirm that the diversity of Campylobacter species on fresh meats is greater than previously reported and highlights the bias of cultural methods towards the recovery of C. jejuni.

  11. The habits of highly effective phages: population dynamics as a framework for identifying therapeutic phages

    Directory of Open Access Journals (Sweden)

    James J Bull

    2014-11-01

    Full Text Available The use of bacteriophages as antibacterial agents is being actively researched on a global scale. Typically, the phages used are isolated from the wild by plating on the bacteria of interest, and a far larger set of candidate phages is often available than can be used in any application. When an excess of phages is available, how should the best phages be identified? Here we consider phage-bacterial population dynamics as a basis for evaluating and predicting phage success. A central question is whether the innate dynamical properties of phages are the determinants of success, or instead, whether extrinsic, indirect effects can be responsible. We address the dynamical perspective, motivated in part by the absence of dynamics in previously suggested principles of phage therapy. Current mathematical models of bacterial-phage dynamics do not capture the realities of in vivo dynamics, nor is this likely to change, but they do give insight to qualitative properties that may be generalizable. In particular, phage adsorption rate may be critical to treatment success, so understanding the effects of the in vivo environment on host availability may allow prediction of useful phages prior to in vivo experimentation. Principles for predicting efficacy may be derived by developing a greater understanding of the in vivo system, or such principles could be determined empirically by comparing phages with known differences in their dynamic properties. The comparative approach promises to be a powerful method of discovering the key to phage success. We offer five recommendations for future study: (i compare phages differing in treatment efficacy to identify the phage properties associated with success, (ii assay dynamics in vivo, (iii understand mechanisms of bacterial escape from phages, (iv test phages in model infections that are relevant to the intended clinical applications, and (v develop new classes of models for phage growth in spatially heterogeneous

  12. Phage inactivation of foodborne Shigella on ready-to-eat spiced chicken.

    Science.gov (United States)

    Zhang, Hui; Wang, Ran; Bao, Hongduo

    2013-01-01

    Shigellosis, also called bacillary dysentery, is an infectious disease caused by Shigella species, including Shigella flexneri, Shigella dysenteriae, Shigella sonnei, and Shigella boydii. Infection with S. flexneri can result in epidemics, and Shigella-contaminated food is often the source of infection, such as ready-to-eat spiced chicken and duck. Therefore, we investigated the ability of Shigella phages to inhibit pathogenic Shigella spp. in ready-to-eat spiced chicken. Food samples were inoculated with individual species (1 × 10(4) cfu/g) or a mixture (S. flexneri 2a, S. dysenteriae, and S. sonnei) to a total concentration of 3 × 10(4) cfu/g. Single phages or a phage cocktail were added thereafter (1 × 10(8) pfu/g or 3 × 10(8) pfu/g), respectively, and samples were incubated at 4°C for 72 h. In general, the application of more phages (3 × 10(8) pfu/g) was the most effective treatment. Phages could reduce bacterial counts by up to 2 log(10)/g after 48 h incubation when treated with the cocktail, and after 72 h the host could not be detected. Similarly, the host in spiced chicken treated with single phage was also sharply reduced after 72 h incubation. The results suggest that an obligately virulent phage cocktail, such as S. flexneri, S. dysenteriae, and S. sonnei phages, can effectively reduce potential contamination of Shigella spp. in ready-to-eat chicken products.

  13. Isolation of Campylobacter spp. from Client-Owned Dogs and Cats, and Retail Raw Meat Pet Food in the Manawatu, New Zealand.

    Science.gov (United States)

    Bojanić, K; Midwinter, A C; Marshall, J C; Rogers, L E; Biggs, P J; Acke, E

    2017-09-01

    Campylobacter causes acute gastroenteritis in people worldwide and is frequently isolated from food, animals and the environment. The disease is predominately food-borne but many routes of transmission and sources of infection have been described, including contact with pets. The prevalence of Campylobacter spp. in dogs and cats varies widely, and data on New Zealand pets are limited. This study aimed to investigate the prevalence of Campylobacter spp. in dogs, cats and retail raw meat pet food products in New Zealand and to characterize Campylobacter jejuni isolates using multilocus sequence typing (MLST). Ninety dogs and 110 cats examined at the Massey University Veterinary Teaching Hospital for elective procedures, and fifty locally purchased retail raw meat pet diets were sampled. Two culture protocols combining Bolton broth enrichment and mCCDA and CAT agars in a microaerobic atmosphere at 42°C and 37°C with species identification using PCR were performed. The prevalence of Campylobacter spp., C. jejuni, Campylobacter upsaliensis and Campylobacter helveticus was 36%, 13%, 23% and 1% in dogs and 16%, 5%, 5% and 7% in cats, respectively. One dog had Campylobacter lari confirmed, and three dogs and one cat had multiple Campylobacter spp. detected. Significantly more animals tested positive using CAT than mCCDA agar (P Campylobacter spp. were isolated from 28%, C. jejuni 22%, C. lari 6% and Campylobacter coli 6% of food samples. Six isolates positive by Campylobacter genus PCR were identified as Arcobacter butzleri. Poultry meat was more likely to be positive than non-poultry meat (P = 0.006). Of the 13 C. jejuni pet isolates with full MLST profiles, eight were of different sequence types (ST) and all nine food isolates were of different STs. © 2016 Blackwell Verlag GmbH.

  14. Subtherapeutic tylosin phosphate in broiler feed affects Campylobacter on carcasses during processing.

    Science.gov (United States)

    Berrang, M E; Ladely, S R; Meinersmann, R J; Fedorka-Cray, P J

    2007-06-01

    Tylosin phosphate is an antimicrobial drug approved for use in broiler feed at subtherapeutic levels for growth promotion. Erythromycin is often the drug of choice for treating humans with campylobacteriosis. Both tylosin and erythromycin are classified as macrolide drugs and cross-resistance between these antimicrobials occurs. Commercial broiler chicks were placed in isolation grow-out chambers and colonized with Campylobacter jejuni. From 14 d of age through grow-out, broilers were fed ad libitim a diet that included 22 ppm of tylosin phosphate (20 g/ton). Control broilers received the same diet without tylosin phosphate. At 42 d of age, broilers were processed in a pilot plant with equipment that closely modeled commercial conditions. Carcass rinses were collected after feather removal, after inside and outside washing, and after immersion chilling. Campylobacter numbers recovered from carcasses after feather removal did not differ according to feed type (3.53 log cfu/mL of rinse for control carcasses, and 3.60 log cfu/mL of rinse for those fed medicated feed). Likewise, medicated feed did not affect Campylobacter numbers on carcasses after inside-outside washing (3.11 and 3.07 log cfu/mL of rinse). However, carcasses of broilers fed tylosin phosphate had lower numbers of Campylobacter after chilling (1.45 log cfu/mL of rinse) than control carcasses (2.31 log cfu/mL of rinse). No Campylobacter isolated from control carcasses were resistant to erythromycin; all Campylobacter recovered from carcasses fed tylosin phosphate were resistant to erythromycin. Application of tylosin phosphate in feed results in lower numbers of Campylobacter on chilled carcasses; however, the Campylobacter that do remain are resistant to erythromycin.

  15. Recent Advances in Screening of Anti-Campylobacter Activity in Probiotics for Use in Poultry

    Science.gov (United States)

    Saint-Cyr, Manuel J.; Guyard-Nicodème, Muriel; Messaoudi, Soumaya; Chemaly, Marianne; Cappelier, Jean-Michel; Dousset, Xavier; Haddad, Nabila

    2016-01-01

    Campylobacteriosis is the most common cause of bacterial gastroenteritis worldwide. Campylobacter species involved in this infection usually include the thermotolerant species Campylobacter jejuni. The major reservoir for C. jejuni leading to human infections is commercial broiler chickens. Poultry flocks are frequently colonized by C. jejuni without any apparent symptoms. Risk assessment analyses have identified the handling and consumption of poultry meat as one of the most important sources of human campylobacteriosis, so elimination of Campylobacter in the poultry reservoir is a crucial step in the control of this foodborne infection. To date, the use of probiotics has demonstrated promising results to reduce Campylobacter colonization. This review provides recent insights into methods used for probiotic screening to reduce the prevalence and colonization of Campylobacter at the farm level. Different eukaryotic epithelial cell lines are employed to screen probiotics with an anti-Campylobacter activity and yield useful information about the inhibition mechanism involved. These in vitro virulence models involve only human intestinal or cervical cell lines whereas the use of avian cell lines could be a preliminary step to investigate mechanisms of C. jejuni colonization in poultry in the presence of probiotics. In addition, in vivo trials to evaluate the effect of probiotics on Campylobacter colonization are conducted, taking into account the complexity introduced by the host, the feed, and the microbiota. However, the heterogeneity of the protocols used and the short time duration of the experiments lead to results that are difficult to compare and draw conclusions at the slaughter-age of broilers. Nevertheless, the combined approach using complementary in vitro and in vivo tools (cell cultures and animal experiments) leads to a better characterization of probiotic strains and could be employed to assess reduced Campylobacter spp. colonization in chickens if some

  16. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the detection of Campylobacter jejuni antibodies, and comparison with a complement fixation test (CFT).

    Science.gov (United States)

    Oosterom, J; den Uyl, C H; Bänffer, J R; Lauwers, S; Huisman, J; Busschbach, A E; Poelma, F G; Bellemans, R

    1985-01-01

    An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of total anti-Campylobacter immunoglobulins in human sera. In this assay disintegrated Campylobacter bacteria were used as the antigen. Absorption tests including other possibly enteropathogenic bacterial species showed that the ELISA system displayed a high immunological specificity for Campylobacter. Using this ELISA it was found that in about 80% of Campylobacter patients these Campylobacter antibodies are produced to almost maximal levels within 8 days after onset of disease, and that they may persist for at least 4 months. Indeed, Campylobacter antibodies were demonstrated at low levels in a large number of control sera. However, accepting an antibody titre of 1:640 as indicative of Campylobacter infection, the statistical sensitivity of the ELISA system was 77% and the specificity 95%. In an epidemiological survey a high association was demonstrated between the severity of Campylobacter-related symptoms and antibody titre values. Assessment of Campylobacter antibody titres by means of this ELISA and by a complement fixation test in 92 sera from index patients and contacts with and without symptoms showed a high association of results.

  17. Temporal variation in the prevalence and species richness of Campylobacter spp. in a prairie watershed impacted by urban and agricultural mixed inputs.

    Science.gov (United States)

    Tambalo, Dinah D; Boa, Tyler; Aryal, Bijaya; Yost, Christopher K

    2016-05-01

    Campylobacter spp. are a substantial cause of gastroenteritis worldwide. Human infection can result from ingestion of contaminated food or water from a variety of sources, including the consumption of fresh produce that is contaminated with the pathogen via the use of contaminated irrigation water. Using molecular methods, we investigated the occurrence of Campylobacter in the Qu'Appelle River watershed, an important source of irrigation water for vegetable producers in southern Saskatchewan, Canada. Water samples were collected from 7 sampling sites from April to September 2009 (145 samples), and from 5 sampling sites from May to October 2013 (116 samples). Campylobacter was detected in 57% and 16% of the samples collected in 2009 and 2013, respectively. Campylobacter detection was highest in May and June for both sampling years. In 2009, the predominant species were Campylobacter lari and Campylobacter jejuni, with prevalences of 84% and 41%, respectively. Other Campylobacter spp. were detected less frequently. Only C. lari was detected in 2013. The results in 2009 demonstrate the species richness of Campylobacter in water sources within the watershed. The occurrence of Campylobacter in the study area also underscores the importance of monitoring irrigation water used to irrigate fresh produce from a public health prospective.

  18. Molecular Subtyping and Source Attribution of Campylobacter Isolated from Food Animals.

    Science.gov (United States)

    Tyson, Gregory H; Tate, Heather P; Abbott, Jason; Tran, Thu-Thuy; Kabera, Claudine; Crarey, Emily; Young, Shenia; McDermott, Patrick F; Sprague, Grisselle; Campbell, Mark; Adeyemo, Oyewole; Browne-Silva, Johnette; Myers, Michael; Thitaram, Sutawee; Zhao, Shaohua

    2016-11-01

    Campylobacter spp. commonly cause gastrointestinal illness in humans. Poultry meats have long been considered the predominant source of these infections, but few in-depth Campylobacter source attribution studies have been completed. We analyzed more than 1,300 Campylobacter isolates recovered from a number of animal and food sources, including dairy and beef cattle, pigs, poultry, and retail poultry meat, and compared them with Campylobacter isolates recovered from human clinical samples. Each isolate was subtyped using pulsed-field gel electrophoresis (PFGE) with SmaI and queried against the Centers for Disease Control and Prevention PulseNet database to identify human isolates with indistinguishable patterns. Half (49.5%) of the PFGE patterns from poultry animal and retail meat isolates were indistinguishable from patterns of at least one human isolate. Among the isolates from beef and dairy cows, 56.6 and 65.0%, respectively, of their PFGE patterns were indistinguishable from those of human isolates. Only a small portion of the PFGE patterns of Campylobacter isolated from pigs (9.5%) were found to have PFGE patterns in common with human isolates. These data imply that cattle may be larger contributors to Campylobacter infections than previously recognized and help further our understanding of potential sources of human campylobacteriosis.

  19. Key role of Mfd in the development of fluoroquinolone resistance in Campylobacter jejuni.

    Directory of Open Access Journals (Sweden)

    Jing Han

    2008-06-01

    Full Text Available Campylobacter jejuni is a major food-borne pathogen and a common causative agent of human enterocolitis. Fluoroquinolones are a key class of antibiotics prescribed for clinical treatment of enteric infections including campylobacteriosis, but fluoroquinolone-resistant Campylobacter readily emerges under the antibiotic selection pressure. To understand the mechanisms involved in the development of fluoroquinolone-resistant Campylobacter, we compared the gene expression profiles of C. jejuni in the presence and absence of ciprofloxacin using DNA microarray. Our analysis revealed that multiple genes showed significant changes in expression in the presence of a suprainhibitory concentration of ciprofloxacin. Most importantly, ciprofloxacin induced the expression of mfd, which encodes a transcription-repair coupling factor involved in strand-specific DNA repair. Mutation of the mfd gene resulted in an approximately 100-fold reduction in the rate of spontaneous mutation to ciprofloxacin resistance, while overexpression of mfd elevated the mutation frequency. In addition, loss of mfd in C. jejuni significantly reduced the development of fluoroquinolone-resistant Campylobacter in culture media or chickens treated with fluoroquinolones. These findings indicate that Mfd is important for the development of fluoroquinolone resistance in Campylobacter, reveal a previously unrecognized function of Mfd in promoting mutation frequencies, and identify a potential molecular target for reducing the emergence of fluoroquinolone-resistant Campylobacter.

  20. Substrate utilization by Campylobacter jejuni and Campylobacter coli

    Energy Technology Data Exchange (ETDEWEB)

    Westfall, H.N.; Rollins, D.M.; Weiss, E.

    1986-10-01

    An attempt was made to elucidate in Campylobacter spp. some of the physiologic characteristics that are reflected in the kinetics of CO/sub 2/ formation from four /sup 14/C-labeled substrates. Campylobacter jejuni and C. coli were grown in a biphasic medium, and highly motile spiral cells were harvested at 12 h. The cells were incubated with 0.02 M glutamate, glutamine, ..cap alpha..-ketoglutarate, or formate, or with concentrations of these substrates ranging from 0.0032 to 0.125 M. All four substrates were metabolized very rapidly by both species. A feature of many of these reactions, particularly obvious with /sup 2/chemically bond-ketoglutarate, was an immediate burst of CO/sub 2/ production followed by CO/sub 2/ evolution at a more moderate rate. These diphasic kinetics of substrate utilization were not seen in comparable experiments with Escherichia coli grown and tested under identical conditions. With C. jejuni, CO/sub 2/ production from formate proceeded rapidly for the entire period of incubation. The rate of metabolism of glutamate, glutamine, and ..cap alpha..-ketoglutarate by both species was greatly enhanced by increased substrate concentration. The approach to the study of the metabolism of campylobacters here described may be useful in detecting subtle changes in the physiology of cells as they are maintained past their logarithmic growth phase.

  1. Monoclonal Antibodies Specific for Hippurate Hydrolase of Campylobacter jejuni

    OpenAIRE

    Steele, Marina; Gyles, Carlton; Chan, Voon Loong; Odumeru, Joseph

    2002-01-01

    Eleven monoclonal antibodies raised against recombinant Campylobacter jejuni hippurate hydrolase were tested for binding to lysates from 19 C. jejuni strains, 12 other Campylobacter strains, and 21 non-Campylobacter strains. Several monoclonal antibodies bound to C. jejuni but not to other Campylobacter species and may be useful in a species-specific immunoassay.

  2. Last of the T Phages

    Energy Technology Data Exchange (ETDEWEB)

    Studier, F. W.

    1978-01-01

    Results clearly show that it is possible to induce mutations in T7 DNA at a physically measurable rate in the laboratory, and to follow genetic divergence by restriction analysis. The rate of accumulation of changes in the presence of mutagen is high enough that it may be feasible to induce changes at least as great as those found among the T7-related phages isolated from nature.

  3. A molecular survey of Campylobacter jejuni and Campylobacter coli virulence and diversity.

    Science.gov (United States)

    Ghorbanalizadgan, Mahdi; Bakhshi, Bita; Kazemnejad Lili, Anoshirvan; Najar-Peerayeh, Shahin; Nikmanesh, Bahram

    2014-07-01

    The aim of this study was to determine the prevalence of virulence-associated genes and enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) analysis of Campylobacter spp. isolated from children with diarrhea in Iran. A total of 200 stool specimens were obtained from children under 5 years during July 2012 to July 2013. Detection of C. jejuni and C. coli was performed by standard biochemical and molecular methods. The presence of virulence-associated genes and genetic diversity of isolates was examined using PCR and ERIC-PCR analyses. A total of 12 (6%) Campylobacter spp. were isolated from patients including 10 (4.5%) C. jejuni and 2 (1.5%) C.coli. The flaA, cadF and ciaB genes were present in 100% of isolates, while no plasmid of virB11 gene was present in their genome. The prevalence of invasion-associated marker was 100% among C. coli and was not detected in C. jejuni isolates. The distribution of both pldA and the genes associated with cytolethal distending toxin (CDT) was 58.3% in C. jejuni isolates. Seven distinct ERIC-PCR profiles were distinguished in three clusters using ERIC-PCR analysis. Genotyping analysis showed a relative correlation with geographic location of patients and virulence gene content of isolates. To our knowledge, this is the first molecular survey of Campylobacter spp. in Iran concerning genotyping and virulence gene content of both C. jejuni and C. coli. ERIC-PCR revealed appropriate discriminatory power for clustering C. jejuni isolates with identical virulence gene content. However, more studies are needed to clearly understand the pathogenesis properties of specific genotypes.

  4. Campylobacter as a venereal disease in cattle

    NARCIS (Netherlands)

    Dijkstra, T.; Wagenaar, J.A.; Visser, I.J.; Bergen, van M.A.P.; Pastoor, P.W.; Strampel, J.; Kock, P.A.

    2005-01-01

    Since the introduction of AI, venereal diseases caused by Tritrichomonas fetus and Campylobacter fetus subsp. venerealis haved been eradicated in The Netherlands. Campylobacter fetus subsp. fetus can cause sporadic abortion and early embryonic death. When natural breeding is practised, venereal dise

  5. Campylobacter jejuni enteritis and reactive arthritis

    OpenAIRE

    Short, C. D.; Klouda, P T; Smith, Lorna

    1982-01-01

    A further case of reactive arthritis following Campylobacter jejuni enteritis is reported. The interim results of a small prospective study are discussed. It may be desirable to do serological studies for campylobacter infection in the investigation of mono- or polyarthritis of acute onset.

  6. Campylobacter iguaniorum sp. nov., isolated from reptiles

    NARCIS (Netherlands)

    Gilbert, Maarten J; Kik, Marja; Miller, William G; Duim, Birgitta; Wagenaar, Jaap A

    2015-01-01

    During samplings of reptiles for Epsilonproteobacteria, Campylobacter strains not belonging to any of the established taxa were isolated from lizards and chelonians. Initial AFLP, PCR, and 16S rRNA sequence analysis showed that these strains were most closely related to Campylobacter fetus and Campy

  7. Flies and Campylobacter infection of broiler flocks

    DEFF Research Database (Denmark)

    Hald, Birthe; Skovgård, Henrik; Bang, Dang Duong

    2004-01-01

    A total of 8.2% of flies caught outside a broiler house in Denmark had the potential to transmit Campylobacter jejuni to chickens, and hundreds of flies per day passed through the ventilation system into the broiler house. Our study suggests that flies may be an important source of Campylobacter...

  8. Quantifying transmission of Campylobacter spp. among broilers

    NARCIS (Netherlands)

    Gerwe, van T.J.; Bouma, A.; Jacobs-Reitsma, W.F.; Broek, van den E.W.F.; Klinkenberg, D.; Stegeman, J.A.; Heesterbeek, J.A.P.

    2005-01-01

    Campylobacter species are frequently identified as a cause of human gastroenteritis, often from eating or mishandling contaminated poultry products. Quantitative knowledge of transmission of Campylobacter in broiler flocks is necessary, as this may help to determine the moment of introduction of

  9. [Campylobacter fetus endocarditis: a case report].

    Science.gov (United States)

    Désidéri-Vaillant, Catherine; Guichon, Jean-Michel; Noyer, Vincent; Nedelec, Yolande; Galinat, Hubert; Sapin-Lory, Jeanne; Di Costanzo, Laurence; Le Guen, Patrick; Nicolas, Xavier

    2013-01-01

    Campylobacter are known to be a cause of enteritidic infections but Campylobacter fetus is more often a cause of systemic infections, mainly in fragilized patients. We report a C. fetus endocarditis. The prognosis seemstobe improved by a prolonged betalactam antibiotic treatment.

  10. Campylobacter in zwemwater en mogelijke emissiebronnen

    NARCIS (Netherlands)

    Ruiter, H.; Rijs, G.; Jacobs-Reitsma, W.F.; Leenen, I.

    2004-01-01

    Campylobacter vormt samen met Salmonella en Shigella één van de belangrijkste bacteriële ziekteverwekkers van het maagdarmkanaal bij de mens. De bacterie Campylobacter komt voor bij eenden, meeuwen, kippen en kalkoenen, maar is ook aanwezig in koeien, varkens en schapen. Al deze dieren zijn drager,

  11. Aspects of epidemiology of Campylobacter in poultry

    NARCIS (Netherlands)

    Jacobs-Reitsma, W.F.

    1997-01-01

    Campylobacter bacteria, which in humans cause infections with severe symptoms of diarrhoea, are mainly transmitted by food, especially poultry meat products. Several studies on Campylobacter colonization in breeders, laying hens, and broilers were carried out. Isolates were serotyped, using a modifi

  12. Aspects of epidemiology of Campylobacter in poultry

    NARCIS (Netherlands)

    Jacobs-Reitsma, W.F.

    1997-01-01

    Campylobacter bacteria, which in humans cause infections with severe symptoms of diarrhoea, are mainly transmitted by food, especially poultry meat products. Several studies on Campylobacter colonization in breeders, laying hens, and broilers were carried out. Isolates were serotyped, using a modifi

  13. Bibliometric analysis of publications on Campylobacter: (2000–2015)

    OpenAIRE

    Sweileh, Waleed M.; Al-Jabi, Samah W.; Sawalha, Ansam F; AbuTaha, Adham S; Zyoud, Sa’ed H.

    2016-01-01

    Background Campylobacter species are widespread zoonotic pathogens. Campylobacter jejuni causes a form of gastroenteritis called campylobacteriosis. Campylobacter drug resistance is considered a serious threat. In order to better understand national and international research output on Campylobacter, we conducted this bibliometric overview of publications on Campylobacter. This study can be used to assess extent of interaction and response of researchers, food regulators, and health policy ma...

  14. Bacteriophages with potential to inactivate Salmonella Typhimurium: Use of single phage suspensions and phage cocktails.

    Science.gov (United States)

    Pereira, Carla; Moreirinha, Catarina; Lewicka, Magdalena; Almeida, Paulo; Clemente, Carla; Cunha, Ângela; Delgadillo, Ivonne; Romalde, Jésus L; Nunes, Maria L; Almeida, Adelaide

    2016-07-15

    The aim of this study was to compare the dynamics of three previously isolated bacteriophages (or phages) individually (phSE-1, phSE-2 and phSE-5) or combined in cocktails of two or three phages (phSE-1/phSE-2, phSE-1/phSE-5, phSE-2/phSE-5 and phSE-1/phSE-2/phSE-5) to control Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) in order to evaluate their potential application during depuration. Phages were assigned to the family Siphoviridae and revealed identical restriction digest profiles, although they showed a different phage adsorption, host range, burst size, explosion time and survival in seawater. The three phages were effective against S. Typhimurium (reduction of ∼2.0 log CFU/mL after 4h treatment). The use of cocktails was not significantly more effective than the use of single phages. A big fraction of the remained bacteria are phage-resistant mutants (frequency of phage-resistant mutants 9.19×10(-5)-5.11×10(-4)) but phage- resistant bacterial mutants was lower for the cocktail phages than for the single phage suspensions and the phage phSE-1 presented the highest rate of resistance and phage phSE-5 the lowest one. The spectral changes of S. Typhimurium resistant and phage-sensitive cells were compared and revealed relevant differences for peaks associated to amide I (1620cm(-1)) and amide II (1515cm(-1)) from proteins and from carbohydrates and phosphates region (1080-1000cm(-1)). Despite the similar efficiency of individual phages, the development of lower resistance indicates that phage cocktails might be the most promising choice to be used during the bivalve depuration to control the transmission of salmonellosis.

  15. Antibody Production in Response to Staphylococcal MS-1 Phage Cocktail in Patients Undergoing Phage Therapy

    OpenAIRE

    Maciej Żaczek; Marzanna Łusiak-Szelachowska; Ewa Jończyk-Matysiak; Beata Weber-Dąbrowska; Ryszard Międzybrodzki; Barbara Owczarek; Agnieszka Kopciuch; Wojciech Fortuna; Paweł Rogóż; Andrzej Górski

    2016-01-01

    In this study, we investigated the humoral immune response (through the release of IgG, IgA, and IgM antiphage antibodies) to a staphylococcal phage cocktail in patients undergoing experimental phage therapy at the Phage Therapy Unit, Medical Center of the Ludwik Hirszfeld Institute of Immunology and Experimental Therapy in Wrocław, Poland. We also evaluated whether occurring antiphage antibodies had neutralizing properties towards applied phages (K rate). Among 20 examined patients receiving...

  16. Complete Genome Sequence of Campylobacter iguaniorum Strain 1485ET, Isolated from a Bearded Dragon (Pogona vitticeps)

    NARCIS (Netherlands)

    Gilbert, Maarten J|info:eu-repo/dai/nl/370804465; Miller, William G; Yee, Emma; Kik, Marja|info:eu-repo/dai/nl/080432565; Wagenaar, Jaap A|info:eu-repo/dai/nl/126613354; Duim, Birgitta|info:eu-repo/dai/nl/143855352

    2014-01-01

    Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to Campylobacter fetus and Campylobacter hyointestinalis. Here we present the first whole-genome sequence for this species.

  17. Complete Genome Sequence of Campylobacter iguaniorum Strain 1485ET, Isolated from a Bearded Dragon (Pogona vitticeps).

    Science.gov (United States)

    Gilbert, Maarten J; Miller, William G; Yee, Emma; Kik, Marja; Wagenaar, Jaap A; Duim, Birgitta

    2014-08-21

    Campylobacter iguaniorum has been isolated from reptiles. This Campylobacter species is genetically related to Campylobacter fetus and Campylobacter hyointestinalis. Here we present the first whole-genome sequence for this species. Copyright © 2014 Gilbert et al.

  18. Campylobacter in poultry, pork and beef

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hasseldam; Carroll, C.; Rudi, K.

    2011-01-01

    Campylobacter infection has become one of the most important zoonoses worldwide. A low prevalence of Campylobacter is generally found in beef and pork at retail, although they may still be sources of infection. Based on the high prevalence of poultry-associated infections, this chapter mainly...... focuses on rapid methods for detection of Campylobacter in this particular production chain, and describes the routes of transmission and sampling in the different levels as well as intervention strategies. The chapter focuses on the introduction, infection dynamics, and sampling of Campylobacter...... throughout the poultry production chain, from farm to consumer level. It also describes culture-based, immunological, and molecular methods for rapid detection, characterization, and enumeration for Campylobacter. Rapid methods can generally be also more sensitive and specific than culture-based methods...

  19. Closely related Campylobacter jejuni strains from different sources reveal a generalist rather than a specialist lifestyle

    Science.gov (United States)

    2011-01-01

    Background Campylobacter jejuni and Campylobacter coli are human intestinal pathogens of global importance. Zoonotic transmission from livestock animals or animal-derived food is the likely cause for most of these infections. However, little is known about their general and host-specific mechanisms of colonization, or virulence and pathogenicity factors. In certain hosts, Campylobacter species colonize persistently and do not cause disease, while they cause acute intestinal disease in humans. Results Here, we investigate putative host-specificity using phenotypic characterization and genome-wide analysis of genetically closely related C. jejuni strains from different sources. A collection of 473 fresh Campylobacter isolates from Germany was assembled between 2006 and 2010 and characterized using MLST. A subset of closely related C. jejuni strains of the highly prevalent sequence type ST-21 was selected from different hosts and isolation sources. PCR typing of strain-variable genes provided evidence that some genes differed between these strains. Furthermore, phenotypic variation of these strains was tested using the following criteria: metabolic variation, protein expression patterns, and eukaryotic cell interaction. The results demonstrated remarkable phenotypic diversity within the ST-21 group, which however did not correlate with isolation source. Whole genome sequencing was performed for five ST-21 strains from chicken, human, bovine, and food sources, in order to gain insight into ST-21 genome diversity. The comparisons showed extensive genomic diversity, primarily due to recombination and gain of phage-related genes. By contrast, no genomic features associated with isolation source or host were identified. Conclusions The genome information and phenotypic data obtained in vitro and in a chicken infection model provided little evidence of fixed adaptation to a specific host. Instead, the dominant C. jejuni ST-21 appeared to be characterized by phenotypic

  20. The Effects of Temperature and Innate Immunity on Transmission of Campylobacter jejuni (Campylobacterales: Campylobacteraceae) Between Life Stages of Musca domestica (Diptera: Muscidae)

    DEFF Research Database (Denmark)

    Bahrndorff, Simon; Gill, C.; Lowenberger, C.

    2014-01-01

    The house fly (Musca domestica L.) is a well-established vector of human pathogens, including Campylobacter spp., which can cause infection of broiler chicken flocks, and through contaminated broiler meat can cause outbreaks of campylobacteriosis in humans. We investigated whether Campylobacter j...

  1. Whole-Genome Sequence of Multidrug-Resistant Campylobacter coli Strain COL B1-266, Isolated from the Colombian Poultry Chain.

    Science.gov (United States)

    Bernal, Johan F; Donado-Godoy, Pilar; Arévalo, Alejandra; Duarte, Carolina; Realpe, María E; Díaz, Paula L; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David; Mariño-Ramírez, Leonardo

    2016-03-17

    Campylobacter coli is considered one of the main causes of food-borne illness worldwide. We report here the whole-genome sequence of multidrug-resistant Campylobacter coli strain COL B1-266, isolated from the Colombian poultry chain. The genome sequences encode genes for a variety of antimicrobial resistance genes, including aminoglycosides, β-lactams, lincosamides, fluoroquinolones, and tetracyclines.

  2. Information Phage Therapy Research Should Report.

    Science.gov (United States)

    Abedon, Stephen T

    2017-04-30

    Bacteriophages, or phages, are viruses which infect bacteria. A large subset of phages infect bactericidally and, consequently, for nearly one hundred years have been employed as antibacterial agents both within and outside of medicine. Clinically these applications are described as phage or bacteriophage therapy. Alternatively, and especially in the treatment of environments, this practice instead may be described as a phage-mediated biocontrol of bacteria. Though the history of phage therapy has involved substantial clinical experimentation, current standards along with drug regulations have placed a premium on preclinical approaches, i.e., animal experiments. As such, it is important for preclinical experiments not only to be held to high standards but also to be reported in a manner which improves translation to clinical utility. Here I address this latter issue, that of optimization of reporting of preclinical as well as clinical experiments. I do this by providing a list of pertinent information and data which, in my opinion, phage therapy experiments ought to present in publications, along with tips for best practices. The goal is to improve the ability of readers to gain relevant information from reports on phage therapy research, to allow other researchers greater potential to repeat or extend findings, to ease transitions from preclinical to clinical development, and otherwise simply to improve phage therapy experiments. Targeted are not just authors but also reviewers, other critical readers, writers of commentaries, and, perhaps, formulators of guidelines or policy. Though emphasizing therapy, many points are applicable to phage-mediated biocontrol of bacteria more generally.

  3. Are host characteristics or exposure factors mainly involved in the acquisition of zoonotic Salmonella and Campylobacter coinfection in humans?

    Science.gov (United States)

    Gradel, Kim O; Schønheyder, Henrik C; Kristensen, Brian; Dethlefsen, Claus; Ejlertsen, Tove; Nielsen, Henrik

    2009-03-01

    We hypothesized that patients coinfected with zoonotic Salmonella and Campylobacter were frailer than monoinfected Salmonella or Campylobacter patients. The study cohort included all first-time Salmonella/Campylobacter infections in Aarhus and North Jutland counties, Denmark, from 1991 through 2003. Data on comorbidity, hospitalization in relation to the Salmonella/Campylobacter infection, and 1-year mortality were obtained from electronic registries. Among 13,449 individuals, 114 (0.85%) had Salmonella/Campylobacter coinfection, 6567 (48.8%) had Salmonella monoinfection, and 6768 (50.3%) had Campylobacter monoinfection. There were no major differences in age, gender, comorbidity, hospitalization rates, 1-year mortality, or seasonal variation between coinfected patients on the one hand and each of the monoinfected patient groups on the other. The main difference was encountered between the Salmonella serotype distribution as 49.1% of coinfected patients versus 20.3% of monoinfected Salmonella patients had Salmonella serotypes other than Salmonella Enteritidis and Salmonella Typhimurium (odds ratio [95% confidence interval]: 4.07 [2.73-6.06]). In conclusion, Salmonella/Campylobacter coinfected patients were not frailer than monoinfected patients. The difference in Salmonella serotype distribution was compatible with a higher proportion of coinfections acquired during foreign travel.

  4. Campylobacter spp. Prevalence and Levels in Raw Milk: A Systematic Review and Meta-Analysis.

    Science.gov (United States)

    Christidis, T; Pintar, K D M; Butler, A J; Nesbitt, A; Thomas, M K; Marshall, B; Pollari, F

    2016-10-01

    Campylobacteriosis is the leading bacterial gastrointestinal disease internationally, contributing significantly to the enteric illness burden. Cases have been associated with the consumption of raw milk, a behavior that has garnered attention recently. Estimates of the prevalence and levels of Campylobacter spp. in raw milk are lacking, which hinders risk assessment attempts. This article is a systematic review and meta-analysis of reported prevalence and levels of zoonotic Campylobacter spp. in the raw milk of cows, goats, and sheep in Canada, the United States, Europe, Australia, and New Zealand. The relevant literature was reviewed, and trained reviewers examined the results for inclusion of articles in the meta-analysis. Relevant data (prevalence and/or level of Campylobacter in raw milk, country of origin, animal species, sample source, Campylobacter species identified, etc.) were extracted, and a meta-analysis was performed in Stata v. 12 (Metaprop command). The weighted mean prevalence of Campylobacter spp. in raw milk samples was 1.18%. Subgroup analyses were conducted to examine how prevalence varied by study characteristics, with the highest prevalence values in studies from the United Kingdom (by country, 6.4%), about cows (by animal species, 1.3%), and including samples taken from inline filters (by sample source, 1.75%) and in studies that included species that are not pathogenic to humans (by Campylobacter species, 1.14%). Two articles each included a single Campylobacter level, 0.16 ± 0.3 and approximately 0.047 most probable number per ml. Despite a relatively low prevalence, consumption of raw milk is inherently risky because no treatment has been used to inactivate pathogens. This potential risk further supports maintaining regulations to limit the sales of raw milk.

  5. Identification of Soft Matter Binding Peptide Ligands Using Phage Display.

    Science.gov (United States)

    Günay, Kemal Arda; Klok, Harm-Anton

    2015-10-21

    Phage display is a powerful tool for the selection of highly affine, short peptide ligands. While originally primarily used for the identification of ligands to proteins, the scope of this technique has significantly expanded over the past two decades. Phage display nowadays is also increasingly applied to identify ligands that selectively bind with high affinity to a broad range of other substrates including natural and biological polymers as well as a variety of low-molecular-weight organic molecules. Such peptides are of interest for various reasons. The ability to selectively and with high affinity bind to the substrate of interest allows the conjugation or immobilization of, e.g., nanoparticles or biomolecules, or generally, facilitates interactions at materials interfaces. On the other hand, presentation of peptide ligands that selectively bind to low-molecular-weight organic materials is of interest for the development of sensor surfaces. The aim of this article is to highlight the opportunities provided by phage display for the identification of peptide ligands that bind to synthetic or natural polymer substrates or to small organic molecules. The article will first provide an overview of the different peptide ligands that have been identified by phage display that bind to these "soft matter" targets. The second part of the article will discuss the different characterization techniques that allow the determination of the affinity of the identified ligands to the respective substrates.

  6. CRISPR: new horizons in phage resistance and strain identification.

    Science.gov (United States)

    Barrangou, Rodolphe; Horvath, Philippe

    2012-01-01

    Bacteria have been widely used as starter cultures in the food industry, notably for the fermentation of milk into dairy products such as cheese and yogurt. Lactic acid bacteria used in food manufacturing, such as lactobacilli, lactococci, streptococci, Leuconostoc, pediococci, and bifidobacteria, are selectively formulated based on functional characteristics that provide idiosyncratic flavor and texture attributes, as well as their ability to withstand processing and manufacturing conditions. Unfortunately, given frequent viral exposure in industrial environments, starter culture selection and development rely on defense systems that provide resistance against bacteriophage predation, including restriction-modification, abortive infection, and recently discovered CRISPRs (clustered regularly interspaced short palindromic repeats). CRISPRs, together with CRISPR-associated genes (cas), form the CRISPR/Cas immune system, which provides adaptive immunity against phages and invasive genetic elements. The immunization process is based on the incorporation of short DNA sequences from virulent phages into the CRISPR locus. Subsequently, CRISPR transcripts are processed into small interfering RNAs that guide a multifunctional protein complex to recognize and cleave matching foreign DNA. Hypervariable CRISPR loci provide insights into the phage and host population dynamics, and new avenues for enhanced phage resistance and genetic typing and tagging of industrial strains.

  7. Genetic Evidence for O-Specific Antigen as Receptor of Pseudomonas aeruginosa Phage K8 and Its Genomic Analysis

    Directory of Open Access Journals (Sweden)

    Xuewei ePan

    2016-03-01

    Full Text Available Phage therapy requires the comprehensive understanding of the mechanisms underlying the host-phage interactions. In this work, to identify the genes related to Pseudomonas aeruginosa phage K8 receptor synthesis, 16 phage-resistant mutants were selected from a Tn5G transposon mutant library of strain PAK. The disrupted genetic loci were identified and they were related to O-specific antigen (OSA synthesis, including gene wbpR, ssg, wbpV, wbpO, and Y880_RS05480, which encoded a putative O-antigen polymerase Wzy. The LPS profile of the Y880_RS05480 mutant was analyzed and shown to lack the O-antigen. Therefore, the data from characterization of Y880_RS05480 by TMHMM and SDS-PAGE silver staining analysis suggest that this locus might encode Wzy. The complete phage K8 genome was characterized as 93879 bp in length and contained identical 1188-bp terminal direct repeats. Comparative genomic analysis showed that phage K8 was highly homologous to members of the genus PaP1-like phages. On the basis of our genetic findings, OSA of P. aeruginosa PAK is proven to be the receptor of phage K8. The highly conserved structural proteins among the genetic closely related phages suggest that they may recognize the same receptor.

  8. Two Novel Myoviruses from the North of Iraq Reveal Insights into Clostridium difficile Phage Diversity and Biology

    Directory of Open Access Journals (Sweden)

    Srwa J. Rashid

    2016-11-01

    Full Text Available Bacteriophages (phages are increasingly being explored as therapeutic agents to combat bacterial diseases, including Clostridium difficile infections. Therapeutic phages need to be able to efficiently target and kill a wide range of clinically relevant strains. While many phage groups have yet to be investigated in detail, those with new and useful properties can potentially be identified when phages from newly studied geographies are characterised. Here, we report the isolation of C. difficile phages from soil samples from the north of Iraq. Two myoviruses, CDKM15 and CDKM9, were selected for detailed sequence analysis on the basis of their broad and potentially useful host range. CDKM9 infects 25/80 strains from 12/20 C. difficile ribotypes, and CDKM15 infects 20/80 strains from 9/20 ribotypes. Both phages can infect the clinically relevant ribotypes R027 and R001. Phylogenetic analysis based on whole genome sequencing revealed that the phages are genetically distinct from each other but closely related to other long-tailed myoviruses. A comparative genomic analysis revealed key differences in the genes predicted to encode for proteins involved in bacterial infection. Notably, CDKM15 carries a clustered regularly interspaced short palindromic repeat (CRISPR array with spacers that are homologous to sequences in the CDKM9 genome and of phages from diverse localities. The findings presented suggest a possible shared evolutionary past for these phages and provides evidence of their widespread dispersal.

  9. Isolation and Identification of Campylobacter spp. from Poultry and Poultry By-Products in Tunisia by Conventional Culture Method and Multiplex Real-Time PCR.

    Science.gov (United States)

    Jribi, Hela; Sellami, Hanen; Mariam, Siala; Smaoui, Salma; Ghorbel, Asma; Hachicha, Salma; Benejat, Lucie; Messadi-Akrout, Feriel; Mégraud, Francis; Gdoura, Radhouane

    2017-10-01

    Thermophilic Campylobacter spp. are one of the primary causes of bacterial human diarrhea. The consumption of poultry meats, by-products, or both is suspected to be a major cause of human campylobacteriosis. The aims of this study were to determine the prevalence of thermophilic Campylobacter spp. in fresh poultry meat and poultry by-products by conventional culture methods and to confirm Campylobacter jejuni and Campylobacter coli isolates by using the multiplex PCR assay. Two hundred fifty fresh poultry samples were collected from a variety of supermarkets and slaughterhouses located in Sfax, Tunisia, including chicken (n =149) and turkey (n =101). The samples were analyzed using conventional microbiological examinations according to the 2006 International Organization for Standardization method (ISO 10272-1) for Campylobacter spp. Concurrently, a real-time PCR was used for identification of C. jejuni and C. coli . Of the 250 samples of poultry meat and poultry by-products, 25.6% (n = 64) were contaminated with Campylobacter spp. The highest prevalence of Campylobacter spp. was found in chicken meat (26.8%) followed by turkey meat (23.7%). Among the different products, poultry breasts showed the highest contamination (36.6%) followed by poultry by-products (30%), poultry wings (28%) and poultry legs (26%) showed the lowest contamination, and no contamination was found on neck skin. Of the 64 thermophilic Campylobacter isolates, C. jejuni (59.7%) was the most frequently isolated species and 10.9% of the isolates were identified as C. coli . All of the 64 Campylobacter isolates identified by the conventional culture methods were further confirmed by PCR. The seasonal peak of Campylobacter spp. contamination was in the warm seasons (spring and summer). The study concluded that high proportions of poultry meat and poultry by-products marketed in Tunisia are contaminated by Campylobacter spp. Furthermore, to ensure food safety, poultry meats must be properly cooked

  10. Exploring the Secretomes of Microbes and Microbial Communities Using Filamentous Phage Display

    Directory of Open Access Journals (Sweden)

    Dragana eGagic

    2016-04-01

    Full Text Available Microbial surface and secreted proteins (the secretome contain a large number of proteins that interact with other microbes, host and/or environment. These proteins are exported by the coordinated activities of the protein secretion machinery present in the cell. A group of phage, called filamentous phage, have the ability to hijack the cellular protein secretion machinery in order to amplify and assemble via a secretion-like process. This ability has been harnessed in the use of filamentous phage of Escherichia coli in biotechnology applications, including screening large libraries of variants for binding to bait of interest, from tissues in vivo to pure proteins or even inorganic substrates. In this review we discuss the roles of secretome proteins in pathogenic and non-pathogenic bacteria and corresponding secretion pathways. We describe the basics of phage display technology and its variants applied to discovery of bacterial proteins that have functions of interest for bacterial colonization and pathogenesis, through filamentous phage display library screening. Published literature also shows that phage display is suitable for secretome protein display as a tool for identification immunogenic peptides and can be used for discovery of vaccine candidates. Secretome selection aided by next-generation sequence analysis can also be used for selective display of the secretome at a microbial community scale, the latter revealing the richness of secretome functions of interest and surprising versatility in filamentous phage display of secretome proteins from large number of Gram-negative as well as Gram-positive bacteria and archaea.

  11. Evidence of udder excretion of Campylobacter jejuni as the cause of milk-borne campylobacter outbreak.

    OpenAIRE

    Hutchinson, D N; Bolton, F. J.; Hinchliffe, P. M.; Dawkins, H. C.; Horsley, S D; Jessop, E G; Robertshaw, P. A.; Counter, D. E.

    1985-01-01

    We describe a community outbreak of campylobacter enteritis associated with the consumption of untreated milk, apparently contaminated by two cows with campylobacter mastitis. The outbreak occurred in two phases. Strains of Campylobacter jejuni of the Penner serogroup complex 4, 13, 16, 50 and Preston biotype code 6100 were isolated from patients in both episodes and from the faeces of the cattle, milk filters, bulk milk and retail milk. Milk samples from two of 40 milking cows were found to ...

  12. Selection of phages and conditions for the safe phage therapy against Pseudomonas aeruginosa infections.

    Science.gov (United States)

    Krylov, Victor; Shaburova, Olga; Pleteneva, Elena; Krylov, Sergey; Kaplan, Alla; Burkaltseva, Maria; Polygach, Olga; Chesnokova, Elena

    2015-02-01

    The emergence of multidrug-resistant bacterial pathogens forced us to consider the phage therapy as one of the possible alternative approaches to treatment. The purpose of this paper is to consider the conditions for the safe, long-term use of phage therapy against various infections caused by Pseudomonas aeruginosa. We describe the selection of the most suitable phages, their most effective combinations and some approaches for the rapid recognition of phages unsuitable for use in therapy. The benefits and disadvantages of the various different approaches to the preparation of phage mixtures are considered, together with the specific conditions that are required for the safe application of phage therapy in general hospitals and the possibilities for the development of personalized phage therapy.

  13. Phage-Phagocyte Interactions and Their Implications for Phage Application as Therapeutics

    Directory of Open Access Journals (Sweden)

    Ewa Jończyk-Matysiak

    2017-06-01

    Full Text Available Phagocytes are the main component of innate immunity. They remove pathogens and particles from organisms using their bactericidal tools in the form of both reactive oxygen species and degrading enzymes—contained in granules—that are potentially toxic proteins. Therefore, it is important to investigate the possible interactions between phages and immune cells and avoid any phage side effects on them. Recent progress in knowledge concerning the influence of phages on phagocytes is also important as such interactions may shape the immune response. In this review we have summarized the current knowledge on phage interactions with phagocytes described so far and their potential implications for phage therapy. The data suggesting that phage do not downregulate important phagocyte functions are especially relevant for the concept of phage therapy.

  14. Selection of phages and conditions for the safe phage therapy against Pseudomonas aeruginosa infections

    Institute of Scientific and Technical Information of China (English)

    Victor; Krylov; Olga; Shaburova; Elena; Pleteneva; Sergey; Krylov; Alla; Kaplan; Maria; Burkaltseva; Olga; Polygach; Elena; Chesnokova

    2015-01-01

    The emergence of multidrug-resistant bacterial pathogens forced us to consider the phage therapy as one of the possible alternative approaches to treatment. The purpose of this paper is to consider the conditions for the safe, long-term use of phage therapy against various infections caused by Pseudomonas aeruginosa. We describe the selection of the most suitable phages, their most effective combinations and some approaches for the rapid recognition of phages unsuitable for use in therapy. The benefi ts and disadvantages of the various different approaches to the preparation of phage mixtures are considered, together with the specifi c conditions that are required for the safe application of phage therapy in general hospitals and the possibilities for the development of personalized phage therapy.

  15. Campylobacter iguaniorum sp. nov., isolated from reptiles.

    Science.gov (United States)

    Gilbert, Maarten J; Kik, Marja; Miller, William G; Duim, Birgitta; Wagenaar, Jaap A

    2015-03-01

    During sampling of reptiles for members of the class Epsilonproteobacteria, strains representing a member of the genus Campylobacter not belonging to any of the established taxa were isolated from lizards and chelonians. Initial amplified fragment length polymorphism, PCR and 16S rRNA sequence analysis showed that these strains were most closely related to Campylobacter fetus and Campylobacter hyointestinalis. A polyphasic study was undertaken to determine the taxonomic position of five strains. The strains were characterized by 16S rRNA and atpA sequence analysis, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry and conventional phenotypic testing. Whole-genome sequences were determined for strains 1485E(T) and 2463D, and the average nucleotide and amino acid identities were determined for these strains. The strains formed a robust phylogenetic clade, divergent from all other species of the genus Campylobacter. In contrast to most currently known members of the genus Campylobacter, the strains showed growth at ambient temperatures, which might be an adaptation to their reptilian hosts. The results of this study clearly show that these strains isolated from reptiles represent a novel species within the genus Campylobacter, for which the name Campylobacter iguaniorum sp. nov. is proposed. The type strain is 1485E(T) ( = LMG 28143(T) = CCUG 66346(T)).

  16. Campylobacter in Poultry: Ecology and Potential Interventions.

    Science.gov (United States)

    Sahin, Orhan; Kassem, Issmat I; Shen, Zhangqi; Lin, Jun; Rajashekara, Gireesh; Zhang, Qijing

    2015-06-01

    Avian hosts constitute a natural reservoir for thermophilic Campylobacter species, primarily Campylobacter jejuni and Campylobacter coli, and poultry flocks are frequently colonized in the intestinal tract with high numbers of the organisms. Prevalence rates in poultry, especially in slaughter-age broiler flocks, could reach as high as 100% on some farms. Despite the extensive colonization, Campylobacter is essentially a commensal in birds, although limited evidence has implicated the organism as a poultry pathogen. Although Campylobacter is insignificant for poultry health, it is a leading cause of food-borne gastroenteritis in humans worldwide, and contaminated poultry meat is recognized as the main source for human exposure. Therefore, considerable research efforts have been devoted to the development of interventions to diminish Campylobacter contamination in poultry, with the intention to reduce the burden of food-borne illnesses. During the past decade, significant advance has been made in understanding Campylobacter in poultry. This review summarizes the current knowledge with an emphasis on ecology, antibiotic resistance, and potential pre- and postharvest interventions.

  17. Campylobacter Enteritis among Children in Iran

    Directory of Open Access Journals (Sweden)

    MR Pourmand

    2005-10-01

    Full Text Available Campylobacter enteritis is a common form of acute gastroenteritis. Among children, especially in developing countries, Campylobacter infections can cause sever life-threatening diarrheal disease. The incidence of Campylobacter infection among children is age related with a higher incidence among younger children in the developing world whereas in industrialized countries the incidence is highest in older children. In a study of American children, Campylobacter was isolated in 4.8% of diarrheal stools in aged 1-4 years. In 1985 the prevalence of Campylobacter diarrheal was 4.4%, whereas in current report 6% of stool samples from children aged<5 years with diarrhea grew Campylobacter jejuni. There were no significant differences between age groups of patients. All thirteen isolated strains of Campylobacter were resistance to Bactrim, Colistin and Polymyxin B and were sensitive to Neomycin, Erythromycin, Gentamicin and Nalidixic acid. The incidence of human campylobacteriosis is increasing worldwide. Thus, public health awareness about the problem is necessary, with a view towards setting up national surveillance programs.

  18. A shortcut in phage screening technique

    Directory of Open Access Journals (Sweden)

    Alexandre de Andrade

    2005-03-01

    Full Text Available A simple modification of the traditional Benton & Davis technique for phage screening is presented that avoids the tedious sample dilutions of putative spots/phages towards the second screening. With the use of a sole agar plate and nylon filter, the modification distinguishes a true positive recombinant from a false positive, with high probability of success.

  19. Methods for Selecting Phage Display Antibody Libraries.

    Science.gov (United States)

    Jara-Acevedo, Ricardo; Diez, Paula; Gonzalez-Gonzalez, Maria; Degano, Rosa Maria; Ibarrola, Nieves; Gongora, Rafael; Orfao, Alberto; Fuentes, Manuel

    2016-01-01

    The selection process aims sequential enrichment of phage antibody display library in clones that recognize the target of interest or antigen as the library undergoes successive rounds of selection. In this review, selection methods most commonly used for phage display antibody libraries have been comprehensively described.

  20. Detection of Campylobacter jejuni and Campylobacter coli in chicken meat samples by real-time nucleic acid sequence-based amplification with molecular beacons.

    Science.gov (United States)

    Churruca, E; Girbau, C; Martínez, I; Mateo, E; Alonso, R; Fernández-Astorga, A

    2007-06-10

    A nucleic acid sequence-based amplification (NASBA) assay based on molecular beacons was used for real-time detection of Campylobacter jejuni and Campylobacter coli in samples of chicken meat. A set of specific primers and beacon probe were designed to target the 16S rRNA of both species. The real-time NASBA protocol including the RNA isolation was valid for both of the cell suspensions in buffered saline and the artificially contaminated chicken meat samples. The presence of rRNA could be correlated with cellular viability, following inactivation of the bacteria by heating, in inoculated chicken meat samples but not in RNase-free cell suspensions.

  1. MAMA-PCR assay for the detection of point mutations associated with high-level erythromycin resistance in Campylobacter jejuni and Campylobacter coli strains.

    Science.gov (United States)

    Alonso, Rodrigo; Mateo, Estibaliz; Churruca, Estibaliz; Martinez, Irati; Girbau, Cecilia; Fernández-Astorga, Aurora

    2005-10-01

    Twenty Campylobacter jejuni and 16 Campylobacter coli strains isolated from humans and food/animals, including 17 isolates resistant to erythromycin, were analyzed. A combined mismatch amplification mutation assay-PCR technique was developed to detect the mutations A 2074 C and A 2075 G in the 23S rRNA gene associated with erythromycin resistance. All high-level erythromycin-resistant strains examined by DNA sequencing carried the transition mutation A 2075 G, whereas no isolate carried the A 2074 C mutation. No mutations were found among the susceptible and low-level erythromycin-resistant strains.

  2. Campylobacter carcass contamination throughout the slaughter process of Campylobacter-positive broiler batches.

    Science.gov (United States)

    Seliwiorstow, Tomasz; Baré, Julie; Van Damme, Inge; Uyttendaele, Mieke; De Zutter, Lieven

    2015-02-02

    Campylobacter contamination on broiler carcasses of Campylobacter colonized flocks was quantified at seven sampling sites throughout the slaughter process. For this purpose, in four slaughterhouses samples were collected from twelve Campylobacter positive batches. Broilers from all visits carried high numbers of campylobacters in their caeca (≥7.9log10cfu/g). Campylobacter counts on feathers (up to 6.8log10cfu/g), positively associated with the breast skin contamination of incoming birds and carcasses after plucking, were identified as an additional source of carcass contamination. A high variability in Campylobacter carcass contamination on breast skin samples within batches and between batches in the same slaughterhouse and between slaughterhouses was observed. In slaughterhouses A, B, C and D Campylobacter counts exceeded a limit of 1000cfu/g on 50%, 56%, 78% and 11% of carcasses after chilling, respectively. This finding indicates that certain slaughterhouses are able to better control Campylobacter contamination than others. Overall, the present study focuses on the descriptive analysis of Campylobacter counts in different slaughterhouses, different batches within a slaughterhouse and within a batch at several sampling locations.

  3. Development of a Novel Chromogenic Medium for Improved Campylobacter Detection from Poultry Samples.

    Science.gov (United States)

    Teramura, Hajime; Iwasaki, Mihoko; Ogihara, Hirokazu

    2015-09-01

    The presence of expanded-spectrum β-lactamase (ESBL)-producing Escherichia coli is a common problem in the isolation of Campylobacter from poultry samples using conventional cefoperazone-based selective media. A novel chromogenic medium (CM-HT), based on modified charcoal cefoperazone deoxycholate agar (mCCDA), has been developed as a solution for improved Campylobacter detection from poultry samples. Although the basic components of CM-HT are the same as mCCDA, CM-HT uses both granular charcoal and sodium cefoxitin to enhance viewability and inhibit ESBL-producing bacteria. All tested Campylobacter jejuni (n = 31) and Campylobacter coli (n = 6) strains grew and formed purple-colored colonies on CM-HT. In contrast, the growth of all other tested microorganisms, including ESBL-producing E. coli strains, was suppressed by this medium. Additionally, 84 poultry samples were examined for the presence of Campylobacter using the ISO 10272-1 method (enrichment with Bolton broth) and the NIHSJ-02 method (enrichment with Preston broth) with mCCDA and CM-HT media for the isolation. The numbers of samples from which Camplylobacter was detected on CM-HT using Preston and Bolton broth were 22 and 18, whereas the numbers on mCCDA were 22 and 13, respectively. Only Campylobacter was detected on CM-HT using both enrichment broths; however, there were 5 and 19 samples from which ESBL-producing E. coli was detected on mCCDA using Preston and Bolton broth, respectively. Thus, there was a significant difference between CM-HT and mCCDA in selectivity for ESBL-producing E. coli regardless of which enrichment broth was used. The results obtained demonstrated that CM-HT is a possible solution for the improved isolation of Campylobacter from poultry samples.

  4. Campylobacter enteritis in adult patients with acute diarrhea from 2005 to 2009 in Beijing, China

    Institute of Scientific and Technical Information of China (English)

    CHEN Jie; SUN Xin-ting; ZENG Zheng; YU Yan-yan

    2011-01-01

    Background There has been a marked global increase in the incidence of human Campylobacter enteritis in recent years. This study investigated the epidemiological and clinical features of Campylobacter enteritis in adult patients suffering from acute diarrhea.Methods This was a retrospective review of Campylobacter enteritis in adult patients with acute diarrhea presenting at Beijing University First Hospital, Beijing, China, in the summer and autumn (April to October) of 2005 to 2009. The data collected included the species of campylobacter identified, and the age, gender, clinical manifestations and results of laboratory test on stool samples collected from the patients. Campylobacter sensitivity tests to various antimicrobial agents were conducted on 80 specimens. Chi-square tests were applied using SPSS13.0 software and a two-sided P value of <0.05 was considered statistically significant.Results Campylobacter spp. isolated from the stool specimens of 142 patients with diarrhea represented 14.9% of all the cases examined. C. jejuni was identified in 127 patients (89.4%) and C. coli in 15 others (10.6%). The infection incidence was highest in the age range of 21-30 years which comprised 21.7% of the total cases examined. Most cases of diarrhea (46 patients) occurred in June. Watery diarrhea (97.2%), abdominal pain (72.5%) and fever (64.8%) were the most common manifestations of enteric campylobacteriosis. Only four patients (2.8%) had bloody diarrhea. The antimicrobial resistance rates were: cefoperazone (100%), levofloxacin (61.3%), gentamicin (12.5%), erythromycin (6.3%), and azithromycin (2.5%).Conclusions Campylobacter was prevalent among adults with acute diarrhea from 2005 to 2009 in Beijing, China. The large number of those afflicted by the disease warrants the commission of a large multicenter study to determine the extent of enteric campylobacteriosis in this region.

  5. Aeromonas phages encode tRNAs for their overused codons.

    Science.gov (United States)

    Prabhakaran, Ramanandan; Chithambaram, Shivapriya; Xia, Xuhua

    2014-01-01

    The GC-rich bacterial species, Aeromonas salmonicida, is parasitised by both GC-rich phages (Aeromonas phages - phiAS7 and vB_AsaM-56) and GC-poor phages (Aeromonas phages - 25, 31, 44RR2.8t, 65, Aes508, phiAS4 and phiAS5). Both the GC-rich Aeromonas phage phiAS7 and Aeromonas phage vB_AsaM-56 have nearly identical codon usage bias as their host. While all the remaining seven GC-poor Aeromonas phages differ dramatically in codon usage from their GC-rich host. Here, we investigated whether tRNA encoded in the genome of Aeromonas phages facilitate the translation of phage proteins. We found that tRNAs encoded in the phage genome correspond to synonymous codons overused in the phage genes but not in the host genes.

  6. Campylobacter ureolyticus: an emerging gastrointestinal pathogen?

    LENUS (Irish Health Repository)

    Bullman, Susan

    2011-03-01

    A total of 7194 faecal samples collected over a 1-year period from patients presenting with diarrhoea were screened for Campylobacter spp. using EntericBio(®) , a multiplex-PCR system. Of 349 Campylobacter-positive samples, 23.8% were shown to be Campylobacter ureolyticus, using a combination of 16S rRNA gene analysis and highly specific primers targeting the HSP60 gene of this organism. This is, to the best of our knowledge, the first report of C. ureolyticus in the faeces of patients presenting with gastroenteritis and may suggest a role for this organism as an emerging enteric pathogen.

  7. The role of radiology in Campylobacter enterocolitis

    Energy Technology Data Exchange (ETDEWEB)

    Demaerel, P.; Ponette, E.; Baert, A.L. (Leuven Univ. (Belgium). Dept. of Radiology); Lacquet, F.; Verbist, L. (Leuven Univ. (Belgium). Dept. of Microbiology); Rutgeerts, A.L. (Leuven Univ. (Belgium). Dept. of Internal Medicine)

    1989-05-01

    A series of 18 patients with diarrhoea and positive stool cultures for Campylobacter jejuni is presented. The most important radiological features were thickening of ileal mucosal folds, of interhaustral indentations and of the ileocaecal valve, lymphoid hyperplasia and microulcerations. Radiology, as well as endoscopy, are both non-specific in Campylobacter jejuni enterocolitis. The importance of radiology is to exclude more typical features of other causes of inflammatory bowel diseases. Moreover, before the result of the stool culture is available, the radiological features should suggest the suspicion of an acute infectious enterocolitis by Campylobacter jejuni as possible diagnosis. (orig.).

  8. Septic abortion caused by Campylobacter jejuni bacteraemia.

    Science.gov (United States)

    Skuhala, Tomislava; Škerk, Višnja; Markotić, Alemka; Bukovski, Suzana; Desnica, Boško

    2016-08-01

    A 20-year-old female patient, 14 weeks pregnant, was admitted to hospital with anamnestic and clinical features of acute pyelonephritis. Clinical signs of septic abortion developed and after obstetric examination the therapy was changed to ampicillin, gentamicin and clindamycin. Campylobacter jejuni was isolated from blood cultures. Pathohistological findings confirmed diagnosis of purulent chorioamnionitis. After 2 weeks of ciprofloxacin administration the patient fully recovered. Campylobacter jejuni was not isolated from stool culture and no signs of acute enteritis were registered during the illness. Invasive forms of Campylobacter disease without enteritis are not unusual in immunocompromised hosts but they are restricted to C. fetus rather than C. jejuni isolates.

  9. Genotypes and antibiotic resistance of canine Campylobacter jejuni isolates.

    Science.gov (United States)

    Amar, Chantal; Kittl, Sonja; Spreng, David; Thomann, Andreas; Korczak, Bożena M; Burnens, André P; Kuhnert, Peter

    2014-01-10

    Campylobacter jejuni is the most important cause of bacterial gastroenteritis in humans. It is a commensal in many wild and domestic animals, including dogs. Whereas genotypes of human and chicken C. jejuni isolates have been described in some detail, only little information on canine C. jejuni genotypes is available. To gain more information on genotypes of canine C. jejuni and their zoonotic potential, isolates from routine diagnostics of diarrheic dogs as well as isolates of a prevalence study in non-diarrheic dogs were analyzed. Prevalence of thermophilic Campylobacter among non-diarrheic dogs was 6.3% for C. jejuni, 5.9% for Campylobacter upsaliensis and 0.7% for Campylobacter coli. The C. jejuni isolates were genotyped by multi locus sequence typing (MLST) and flaB typing. Resistance to macrolides and quinolones was genetically determined in parallel. Within the 134 genotyped C. jejuni isolates 57 different sequence types (ST) were found. Five STs were previously unrecognized. The most common STs were ST-48 (11.2%), ST-45 (10.5%) and ST-21 (6.0%). Whereas no macrolide resistance was found, 28 isolates (20.9%) were resistant to quinolones. ST-45 was significantly more prevalent in diarrheic than in non-diarrheic dogs. Within the common time frame of isolation 94% of the canine isolates had a ST that was also found in human clinical isolates. In conclusion, prevalence of C. jejuni in Swiss dogs is low but there is a large genetic overlap between dog and human isolates. Given the close contact between human and dogs, the latter should not be ignored as a potential source of human campylobacteriosis. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Economics of zoonoses surveillance in a 'One Health' context: an assessment of Campylobacter surveillance in Switzerland.

    Science.gov (United States)

    Babo Martins, S; Rushton, J; Stärk, K D C

    2017-04-01

    Cross-sectorial surveillance and general collaboration between the animal and the public health sectors are increasingly recognized as needed to better manage the impacts of zoonoses. From 2009, the Swiss established a Campylobacter mitigation system that includes human and poultry surveillance data-sharing within a multi-sectorial platform, in a 'One Health' approach. The objective of this study was to explore the economics of this cross-sectorial approach, including surveillance and triggered interventions. Costs and benefits of the One Health and of the uni-sectorial approach to Campylobacter surveillance were identified using an economic assessment framework developed earlier. Cost information of surveillance activities and interventions was gathered and disability-adjusted life years (DALYs) associated with the disease estimated for 2008 and 2013. In the first 5 years of this One Health approach to Campylobacter mitigation, surveillance contributed with information mainly used to perform risk assessments, monitor trends and shape research efforts on Campylobacter. There was an increase in costs associated with the mitigation activities following integration, due mainly to the allocation of additional resources to research and implementation of poultry surveillance. The overall burden of campylobacteriosis increased by 3·4-8·8% to 1751-2852 DALYs in 2013. In the timing of the analysis, added value associated with this cross-sectorial approach to surveillance of Campylobacter in the country was likely generated through non-measurable benefits such as intellectual capital and social capital.

  11. Campylobacter and Salmonella are prevalent in broiler farms in Kyushu, Japan: results of a 2-year distribution and circulation dynamics audit.

    Science.gov (United States)

    Yamazaki, W; Uemura, R; Sekiguchi, S; Dong, J-B; Watanabe, S; Kirino, Y; Mekata, H; Nonaka, N; Norimine, J; Sueyoshi, M; Goto, Y; Horii, Y; Kurogi, M; Yoshino, S; Misawa, N

    2016-06-01

    To elucidate the distribution and circulation dynamics of Campylobacter and Salmonella in Japanese chicken broiler flocks. A 2-year investigation of the distribution of Campylobacter and Salmonella was conducted in 25 broiler flocks at nine farms in Japan from 2013 to 2014. Campylobacter and Salmonella tested positive in 11 (44·0%) and 24 (96·0%) broiler flocks respectively. One hundred and ninety-five Campylobacter and 184 Salmonella isolates were characterized into 12 Campylobacter (including two novel genotypes) and three Salmonella MLST genotypes. Only Salmonella isolation between caecal and environmental samples were significantly correlated. Further, one litter sample tested positive for Salmonella before new chicks were introduced. The Campylobacter strains rapidly lost culturability within 2-18 days; in contrast, the Salmonella strains survived from 64-211 days in artificially inoculated water samples. No persistent circulation-mediated Campylobacter contamination was observed. In contrast, circulation of Salmonella in broiler houses was seen, apparently due to the litter excreted from broiler flocks, as well as Salmonella-contaminated water and feed. This paper provides the distribution, genotypic data and circulation dynamics of Campylobacter and Salmonella as recently observed in Japanese chicken broiler farms. © 2016 The Society for Applied Microbiology.

  12. A cross-sectional study examining Campylobacter and other zoonotic enteric pathogens in dogs that frequent dog parks in three cities in south-western Ontario and risk factors for shedding of Campylobacter spp.

    Science.gov (United States)

    Procter, T D; Pearl, D L; Finley, R L; Leonard, E K; Janecko, N; Reid-Smith, R J; Weese, J S; Peregrine, A S; Sargeant, J M

    2014-05-01

    An estimated 6 million pet dogs live in Canadian households with the potential to transmit zoonotic pathogens to humans. Dogs have been identified as carriers of Salmonella, Giardia and Campylobacter spp., particularly Campylobacter upsaliensis, but little is known about the prevalence and risk factors for these pathogens in pet dogs that visit dog parks. This study examined the prevalence of these organisms in the faeces of dogs visiting dog parks in three cities in south-western Ontario, as well as risk factors for shedding Campylobacter spp. and C. upsaliensis. From May to August 2009, canine faecal samples were collected at ten dog parks in the cities of Guelph and Kitchener-Waterloo, Ontario, Canada. Owners were asked to complete a questionnaire related to pet characteristics and management factors including age, diet and activities in which the dog participates. Faecal samples were collected from 251 dogs, and 189 questionnaires were completed. Salmonella, Giardia and Campylobacter spp. were present in 1.2%, 6.4% and 43.0% of faecal samples, respectively. Of the Campylobacter spp. detected, 86.1% were C. upsaliensis, 13% were C. jejuni and 0.9% were C. coli. Statistically significant sparing factors associated with the shedding of Campylobacter spp. included the feeding of a commercial dry diet and the dog's exposure to compost. Age of dog had a quadratic effect, with young dogs and senior dogs having an increased probability of shedding Campylobacter spp. compared with adult dogs. The only statistically significant risk factor for shedding C. upsaliensis was outdoor water access including lakes and ditches, while dogs >1 year old were at a lower risk than young dogs. Understanding the pet-related risk factors for Campylobacter spp. and C. upsaliensis shedding in dogs may help in the development of awareness and management strategies to potentially reduce the risk of transmitting this pathogen from dogs to humans.

  13. BREX is a novel phage resistance system widespread in microbial genomes.

    Science.gov (United States)

    Goldfarb, Tamara; Sberro, Hila; Weinstock, Eyal; Cohen, Ofir; Doron, Shany; Charpak-Amikam, Yoav; Afik, Shaked; Ofir, Gal; Sorek, Rotem

    2015-01-13

    The perpetual arms race between bacteria and phage has resulted in the evolution of efficient resistance systems that protect bacteria from phage infection. Such systems, which include the CRISPR-Cas and restriction-modification systems, have proven to be invaluable in the biotechnology and dairy industries. Here, we report on a six-gene cassette in Bacillus cereus which, when integrated into the Bacillus subtilis genome, confers resistance to a broad range of phages, including both virulent and temperate ones. This cassette includes a putative Lon-like protease, an alkaline phosphatase domain protein, a putative RNA-binding protein, a DNA methylase, an ATPase-domain protein, and a protein of unknown function. We denote this novel defense system BREX (Bacteriophage Exclusion) and show that it allows phage adsorption but blocks phage DNA replication. Furthermore, our results suggest that methylation on non-palindromic TAGGAG motifs in the bacterial genome guides self/non-self discrimination and is essential for the defensive function of the BREX system. However, unlike restriction-modification systems, phage DNA does not appear to be cleaved or degraded by BREX, suggesting a novel mechanism of defense. Pan genomic analysis revealed that BREX and BREX-like systems, including the distantly related Pgl system described in Streptomyces coelicolor, are widely distributed in ~10% of all sequenced microbial genomes and can be divided into six coherent subtypes in which the gene composition and order is conserved. Finally, we detected a phage family that evades the BREX defense, implying that anti-BREX mechanisms may have evolved in some phages as part of their arms race with bacteria.

  14. Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine

    OpenAIRE

    Nobrega, Franklin L.; Ana Rita Costa; Santos,José F.; Siliakus, Melvin F.; van Lent, Jan W. M.; Kengen, Servé W. M.; Joana Azeredo; Kluskens, Leon D.

    2016-01-01

    Orally administered phages to control zoonotic pathogens face important challenges, mainly related to the hostile conditions found in the gastrointestinal tract (GIT). These include temperature, salinity and primarily pH, which is exceptionally low in certain compartments. Phage survival under these conditions can be jeopardized and undermine treatment. Strategies like encapsulation have been attempted with relative success, but are typically complex and require several optimization steps. He...

  15. Campylobacter in poultry, pork and beef

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hasseldam; Carroll, C.; Rudi, K.

    2011-01-01

    Campylobacter infection has become one of the most important zoonoses worldwide. A low prevalence of Campylobacter is generally found in beef and pork at retail, although they may still be sources of infection. Based on the high prevalence of poultry-associated infections, this chapter mainly foc...... positive samples to produce isolates for further characterization. The choice of a rapid method will always depend on the requested information and be influenced by the relevant matrix and the expected level of contamination....

  16. Phage Display Technology in Biomaterials Engineering: Progress and Opportunities for Applications in Regenerative Medicine.

    Science.gov (United States)

    Martins, Ivone M; Reis, Rui L; Azevedo, Helena S

    2016-11-18

    The field of regenerative medicine has been gaining momentum steadily over the past few years. The emphasis in regenerative medicine is to use various in vitro and in vivo approaches that leverage the intrinsic healing mechanisms of the body to treat patients with disabling injuries and chronic diseases such as diabetes, osteoarthritis, and degenerative disorders of the cardiovascular and central nervous system. Phage display has been successfully employed to identify peptide ligands for a wide variety of targets, ranging from relatively small molecules (enzymes, cell receptors) to inorganic, organic, and biological (tissues) materials. Over the past two decades, phage display technology has advanced tremendously and has become a powerful tool in the most varied fields of research, including biotechnology, materials science, cell biology, pharmacology, and diagnostics. The growing interest in and success of phage display libraries is largely due to its incredible versatility and practical use. This review discusses the potential of phage display technology in biomaterials engineering for applications in regenerative medicine.

  17. Phage-based magnetostrictive-acoustic microbiosensors for detecting bacillus anthracis spores

    Science.gov (United States)

    Wan, J.; Yang, H.; Lakshmanan, R. S.; Guntupalli, R.; Huang, S.; Hu, J.; Petrenko, V. A.; Chin, B. A.

    2006-05-01

    Magnetostrictive particles (MSPs) as biosensor platform have been developed recently. The principle of MSPs as sensor platform is the same as that of other acoustic wave devices, such as quartz crystal microbalance. In this paper, the fabrication, characterization and performance of phage-based MSP biosensors for detecting Bacillus anthracis spores are reported. A commercially available magnetostrictive alloy was utilized to fabricate the sensor platform. The phage was immobilized onto the MSPs using physical adsorption technology. The following performance of the phage-based MSP sensors will be presented: sensitivity, response time, longevity, specificity and binding efficacy. The performance of the sensors at static and dynamic conditions was characterized. The experimental results are confirmed by microscopy photographs. The excellent performance including high sensitivity and rapid response is demonstrated. More importantly, it is experimentally found that the phage-based MSP sensors have a much better longevity than antibody-based sensors.

  18. Tetracycline consumption and occurrence of tetracycline resistance in Salmonella typhimurium phage types from Danish pigs

    DEFF Research Database (Denmark)

    Emborg, Hanne-Dorthe; Vigre, Håkan; Jensen, Vibeke Frøkjær;

    2007-01-01

    The aims of the present study were to investigate at the farm-owner level the effect of prescribed tetracycline consumption in pigs and different Salmonella Typhimurium phage types on the probability that the S. Typhimurium was resistant to tetracycline. In this study, 1,307 isolates were included......, originating from 877 farm owners, and data were analyzed using logistic regression. The analysis showed that both the S. Typhimurium phage type (p consumption (p = 0.0007) were significantly associated with tetracycline resistance. In particular, the phage type...... was strongly associated with tetracycline resistance. A further analysis of data from the Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) indicates that the tetracycline-susceptible phage types only slowly become tetracycline resistant, although tetracycline consumption...

  19. Symptomatic and Asymptomatic Campylobacter Infections Associated with Reduced Growth in Peruvian Children

    Science.gov (United States)

    Lee, Gwenyth; Pan, William; Peñataro Yori, Pablo; Paredes Olortegui, Maribel; Tilley, Drake; Gregory, Michael; Oberhelman, Richard; Burga, Rosa; Chavez, Cesar Banda; Kosek, Margaret

    2013-01-01

    Background Although diarrheal illnesses are recognized as both a cause and effect of undernutrition, evidence for the effect of specific enteropathogens on early childhood growth remains limited. We estimated the effects of undernutrition as a risk factor for campylobacteriosis, as well as associations between symptomatic and asymptomatic Campylobacter infections and growth. Methodology/Principal Findings Using data from a prospective cohort of 442 children aged 0–72 months, the effect of nutritional status on the incidence of Campylobacter infection was estimated using uni- and multivariate Poisson models. Multivariate regression models were developed to evaluate the effect of Campylobacter infection on weight gain and linear growth. Overall, 8.3% of diarrheal episodes were associated with Campylobacter (crude incidence rate = 0.37 episodes/year) and 4.9% of quarterly asymptomatic samples were Campylobacter positive. In univariate models, the incidence of Campylobacter infection was marginally higher in stunted than non-stunted children (IRR 1.270, 95% CI (0.960, 1.681)(p = 0.095). When recent diarrheal burdens were included in the analysis, there was no difference in risk between stunted and unstunted children. Asymptomatic and symptomatic Campylobacter infections were associated with reduced weight gain over a three-month period (65.5 g (95% CI: −128.0, −3.0)(p = 0.040) and 43.9 g (95% CI:−87.6, −1.0)(p = 0.049) less weight gain, respectively). Symptomatic Campylobacter infections were only marginally associated with reduced linear growth over a nine month period (−0.059 cm per episode, 95% CI: −0.118, 0.001)(p = 0.054), however relatively severe episodes were associated with reduced linear growth (−0.169 cm/episode, 95% CI −0.310, −0.028)(p = 0.019). Conclusions/Significance Our findings suggest that Campylobacter is not as benign as commonly assumed, and that there is evidence to support expanding the indications for

  20. Information Phage Therapy Research Should Report

    Directory of Open Access Journals (Sweden)

    Stephen T. Abedon

    2017-04-01

    Full Text Available Bacteriophages, or phages, are viruses which infect bacteria. A large subset of phages infect bactericidally and, consequently, for nearly one hundred years have been employed as antibacterial agents both within and outside of medicine. Clinically these applications are described as phage or bacteriophage therapy. Alternatively, and especially in the treatment of environments, this practice instead may be described as a phage-mediated biocontrol of bacteria. Though the history of phage therapy has involved substantial clinical experimentation, current standards along with drug regulations have placed a premium on preclinical approaches, i.e., animal experiments. As such, it is important for preclinical experiments not only to be held to high standards but also to be reported in a manner which improves translation to clinical utility. Here I address this latter issue, that of optimization of reporting of preclinical as well as clinical experiments. I do this by providing a list of pertinent information and data which, in my opinion, phage therapy experiments ought to present in publications, along with tips for best practices. The goal is to improve the ability of readers to gain relevant information from reports on phage therapy research, to allow other researchers greater potential to repeat or extend findings, to ease transitions from preclinical to clinical development, and otherwise simply to improve phage therapy experiments. Targeted are not just authors but also reviewers, other critical readers, writers of commentaries, and, perhaps, formulators of guidelines or policy. Though emphasizing therapy, many points are applicable to phage-mediated biocontrol of bacteria more generally.

  1. Rapid enumeration of phage in monodisperse emulsions.

    Science.gov (United States)

    Tjhung, Katrina F; Burnham, Sean; Anany, Hany; Griffiths, Mansel W; Derda, Ratmir

    2014-06-17

    Phage-based detection assays have been developed for the detection of viable bacteria for applications in clinical diagnosis, monitoring of water quality, and food safety. The majority of these assays deliver a positive readout in the form of newly generated progeny phages by the bacterial host of interest. Progeny phages are often visualized as plaques, or holes, in a lawn of bacteria on an agar-filled Petri dish; however, this rate-limiting step requires up to 12 h of incubation time. We have previously described an amplification of bacteriophages M13 inside droplets of media suspended in perfluorinated oil; a single phage M13 in a droplet yields 10(7) copies in 3-4 h. Here, we describe that encapsulation of reporter phages, both lytic T4-LacZ and nonlytic M13, in monodisperse droplets can also be used for rapid enumeration of phage. Compartmentalization in droplets accelerated the development of the signal from the reporter enzyme; counting of "positive" droplets yields accurate enumeration of phage particles ranging from 10(2) to 10(6) pfu/mL. For enumeration of T4-LacZ phage, the fluorescent signal appeared in as little as 90 min. Unlike bulk assays, quantification in emulsion is robust and insensitive to fluctuations in environmental conditions (e.g., temperature). Power-free emulsification using gravity-driven flow in the absence of syringe pumps and portable fluorescence imaging solutions makes this technology promising for use at the point of care in low-resource environments. This droplet-based phage enumeration method could accelerate and simplify point-of-care detection of the pathogens for which reporter bacteriophages have been developed.

  2. Better Campylobacter Detection: Furthering our understanding of Campylobacter ecology in poultry

    Science.gov (United States)

    Campylobacter is an important human pathogen and consumption of undercooked poultry has been linked to significant human illnesses. To reduce human illness, intervention strategies targeting Campylobacter reduction in poultry are in development. For more than a decade, there has been an ongoing na...

  3. Influxed insects as Vectors for Campylobacter jejuni and Campylobacter coll in Danish Broiler Houses

    DEFF Research Database (Denmark)

    Hald, Birthe; Skovgård, Henrik; Pedersen, Karl

    2008-01-01

    The vector potential of flies (Diptera: Brachycera) for spread of Campylobacter jejuni and Campylobacter coli on 5 Danish broiler farms was evaluated in a longitudinal field study from April to November 2004. First, the prevalence of C. jejuni- and C. coli-positive flies was determined in 2...

  4. Presence of antibodies against campylobacter flagellar capping proteins versus campylobacter jejuni isolation in broilers

    Science.gov (United States)

    Campylobacter jejuni is the leading foodborne pathogen that causes human acute bacterial gastroenteritis worldwide. Human cases have been linked to consumption and/or handling of contaminated poultry products. Although Campylobacter jejuni is commonly regarded as a commensal in broiler cecal micro...

  5. An outbreak of infectious hepatitis in commercially reared ostriches associated with Campylobacter coli and Campylobacter jejuni

    DEFF Research Database (Denmark)

    Stephens, C.P.; On, S.L.W.; Gibson, J.A.

    1998-01-01

    lesions resembled those of vibrionic hepatitis in other avian species. Campylobacter coli was isolated from the livers of affected ostriches from five of the six properties. Campylobacter jejuni subsp. jejuni was isolated from birds from the remaining property. Pulsed-field gel electrophoresis-based (PFGE...

  6. Risicobeheersing Campylobacter op het vleeskuikenbedrijf = Campylobacter control on the broiler farm

    NARCIS (Netherlands)

    Veldkamp, T.; Bokma-Bakker, M.H.

    2004-01-01

    In dit onderzoek werden de risico’s voor introductie en verspreiding van Campylobacter op het vleeskuikenbedrijf geïnventariseerd. Er worden mogelijkheden aangegeven hoe deze risico’s beheerst kunnen worden. Het blijkt dat er veel mogelijkheden zijn waardoor Campylobacter geïntroduceerd of verspreid

  7. Risicobeheersing Campylobacter op het vleeskuikenbedrijf = Campylobacter control on the broiler farm

    NARCIS (Netherlands)

    Veldkamp, T.; Bokma-Bakker, M.H.

    2004-01-01

    In dit onderzoek werden de risico’s voor introductie en verspreiding van Campylobacter op het vleeskuikenbedrijf geïnventariseerd. Er worden mogelijkheden aangegeven hoe deze risico’s beheerst kunnen worden. Het blijkt dat er veel mogelijkheden zijn waardoor Campylobacter geïntroduceerd of verspreid

  8. Ribosomal operon intergenic sequence region (ISR) heterogeneity in Campylobacter coli and Campylobacter jejuni

    Science.gov (United States)

    Campylobacter jejuni and Campylobacter coli are closely related species that can not be distinguished by their 16S or 23S rRNA gene sequences. However, the intergenic sequence region (ISR) that is between the 16S and 23S genes is markedly different and characteristic for each species. A peculiarit...

  9. Campylobacter bacteremia: a rare and under-reported event?

    Science.gov (United States)

    Louwen, R.; van Baarlen, P.; van Vliet, A. H. M.; van Belkum, A.; Hays, J. P.; Endtz, H. P.

    2012-01-01

    Bacteria belonging to the species Campylobacter are the most common cause of bacterial diarrhoea in humans. The clinical phenotype associated with Campylobacter infections ranges from asymptomatic conditions to severe colitis and bacteremia. In susceptible patients, Campylobacter infections are associated with significant morbidity and mortality, with both host factors and bacterial factors being involved in the pathogenesis of bacteremia. In the host, age, gender and immune-compromising conditions may predispose for Campylobacter infections, whilst the most important bacterial determinants mentioned in the literature are cytotoxin production and flagellar motility. The role of sialylated lipo-oligosaccharide (LOS) and serum resistance in bacteremia is inconclusive at this time, and the clinical significance of Campylobacter bacteremia is not yet fully understood. More emphasis on the detection of Campylobacter species from blood cultures in susceptible patients at risk for Campylobacter infections will increase our understanding of the pathogenesis and the relevance of Campylobacter bacteremia. PMID:24611124

  10. Selective medium for growth of Campylobacter in containers incubated aerobically

    Science.gov (United States)

    Introduction. Campylobacter are traditionally cultured in primary containers inside of secondary containers filled with microaerobic atmospheres. Recent findings indicated that media supplemented with optimal concentrations of amino acids, organic acids, and bicarbonate support Campylobacter growth ...

  11. The prevalence of thermotolerant Campylobacter species in food ...

    African Journals Online (AJOL)

    Preferred Customer

    The prevalence of thermotolerant Campylobacter species in food animals ... However, little is known about the presence of campylobacter bacteria in various food animals as possible ..... humans, which demonstrates the importance of chicken.

  12. Mammalian Host-Versus-Phage immune response determines phage fate in vivo

    OpenAIRE

    Katarzyna Hodyra-Stefaniak; Paulina Miernikiewicz; Jarosław Drapała; Marek Drab; Ewa Jończyk-Matysiak; Dorota Lecion; Zuzanna Kaźmierczak; Weronika Beta; Joanna Majewska; Marek Harhala; Barbara Bubak; Anna Kłopot; Andrzej Górski; Krystyna Dąbrowska

    2015-01-01

    Emerging bacterial antibiotic resistance draws attention to bacteriophages as a therapeutic alternative to treat bacterial infection. Examples of phage that combat bacteria abound. However, despite careful testing of antibacterial activity in vitro, failures nevertheless commonly occur. We investigated immunological response of phage antibacterial potency in vivo. Anti-phage activity of phagocytes, antibodies, and serum complement were identified by direct testing and by high-resolution fluor...

  13. Prevalence of thermotolerant Campylobacter spp. in farmed hares (Lepus europaeus).

    Science.gov (United States)

    Santaniello, Antonio; Dipineto, Ludovico; Veneziano, Vincenzo; Mariani, Ugo; Fioretti, Alessandro; Menna, Lucia Francesca

    2014-10-01

    Thermotolerant Campylobacter spp. were isolated from 118/240 (49.2%) rectal swabs from commercially farmed hares (Lepus europaeus) in southern Italy. Using multiplex PCR, Campylobacter coli was identified in 118/118 (100%) positive samples, while 17/118 (14.4%) positive samples were also positive for Campylobacter jejuni. Adult hares had a higher prevalence of infection with Campylobacter spp. than juvenile hares.

  14. The effect of slaughter operations on the contamination of chicken carcasses with thermotolerant Campylobacter

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Sommer, Helle Mølgaard; Nielsen, Niels L.;

    2006-01-01

    concentration of 0.5 log(10) cfu/g in average, whereas no significant changes were observed during this operation in Plant II. Air chilling (Plant 1) and water chilling (Plant 11), both including a carcass wash prior to the chilling operation, caused similar, but significant reductions of 0.83 and 0.97 log(10......) cfu/g, respectively. In packed frozen chickens (Plant II) an additional reduction of 1.38 log(10) cfu/g in average was obtained due to the freezing operation. In packed chilled chickens (Plant 1), however, the number of thermotolerant Campylobacter per gram remained at the same level as after air......) and had intestinal mean counts ranging from 6.65 to 8.20 log(10) cfu/g. A correlation between Campylobacter concentrations in intestinal content and on chicken carcasses after the defeathering operation was documented. This finding indicates that a reduction in the Campylobacter concentration on chicken...

  15. High-resolution genomic fingerprinting of Campylobacter jejuni and Campylobacter coli by analysis of amplified fragment length polymorphisms

    DEFF Research Database (Denmark)

    Kokotovic, Branko; On, Stephen L.W.

    1999-01-01

    A method for high-resolution genomic fingerprinting of the enteric pathogens Campylobacter jejuni and Campylobacter coli, based on the determination of amplified fragment length polymorphism, is described. The potential of this method for molecular epidemiological studies of these species...

  16. 21 CFR 866.3110 - Campylobacter fetus serological reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Campylobacter fetus serological reagents. 866.3110... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3110 Campylobacter fetus serological reagents. (a) Identification. Campylobacter fetus serological reagents are devices...

  17. Proteomic and genomic analysis reveals novel Campylobacter jejuni outer membrane proteins and potential heterogeneity.

    Science.gov (United States)

    Watson, Eleanor; Sherry, Aileen; Inglis, Neil F; Lainson, Alex; Jyothi, Dushyanth; Yaga, Raja; Manson, Erin; Imrie, Lisa; Everest, Paul; Smith, David G E

    2014-09-01

    Gram-negative bacterial outer membrane proteins play important roles in the interaction of bacteria with their environment including nutrient acquisition, adhesion and invasion, and antibiotic resistance. In this study we identified 47 proteins within the Sarkosyl-insoluble fraction of Campylobacter jejuni 81-176, using LC-ESI-MS/MS. Comparative analysis of outer membrane protein sequences was visualised to reveal protein distribution within a panel of Campylobacter spp., identifying several C. jejuni-specific proteins. Smith-Waterman analyses of C. jejuni homologues revealed high sequence conservation amongst a number of hypothetical proteins, sequence heterogeneity of other proteins and several proteins which are absent in a proportion of strains.

  18. Genome Sequence of Mycobacterium Phage Waterfoul

    Science.gov (United States)

    Jackson, Paige N.; Embry, Ella K.; Johnson, Christa O.; Watson, Tiara L.; Weast, Sayre K.; DeGraw, Caroline J.; Douglas, Jessica R.; Sellers, J. Michael; D’Angelo, William A.

    2016-01-01

    Waterfoul is a newly isolated temperate siphovirus of Mycobacterium smegmatis mc2155. It was identified as a member of the K5 cluster of Mycobacterium phages and has a 61,248-bp genome with 95 predicted genes. PMID:27856585

  19. The globins of Campylobacter jejuni.

    Science.gov (United States)

    Tinajero-Trejo, Mariana; Shepherd, Mark

    2013-01-01

    Campylobacter jejuni is a zoonotic Gram-negative bacterial pathogen that is exposed to reactive nitrogen species, such as nitric oxide, from a variety of sources. To combat the toxic effects of this nitrosative stress, C. jejuni upregulates a small regulon under the control of the transcriptional activator NssR, which positively regulates the expression of a single-domain globin protein (Cgb) and a truncated globin protein (Ctb). Cgb has previously been shown to detoxify nitric oxide, but the role of Ctb remains contentious. As C. jejuni is amenable to genetic manipulation, and its globin proteins are easily expressed and purified, a combination of mutagenesis, complementation, transcriptomics, spectroscopic characterisation and structural analyses has been used to probe the regulation, function and structure of Cgb and Ctb. This ability to study Cgb and Ctb with such a multi-pronged approach is a valuable asset, especially since only a small fraction of known globin proteins have been functionally characterised.

  20. Multi drug resistance of campylobacter jejuni and campylobacter coli to tested antibiotics in strains originating from humans, poultry and swine

    Directory of Open Access Journals (Sweden)

    Tambur Zoran Ž.

    2010-01-01

    Full Text Available Thermophilic Campylobacter are among the most common cause of bacterial enteritis in humans. Food animals are considered one of the most important sources of Campylobacter causing infections in man. Campylobacter infection is clinically mild and resolves spontaneously. In severe or long-lasting cases, treatment with antibiotics is necessary. Resistance of Campylobacter spp. to drugs used in treatment of infection is a matter of concern. The aim of this paper is to determine presence of multi drug resistant strains of Campylobacter jejuni and Campylobacter coli isolated from animals and man. Material for testing was obtained by scraping the cecum surface from boilers, pig cecum and colon, and human feces. For isolation Campylobacter jejuni and Campylobacter coli microaerophilic conditions, temperature of 42°C and antibiotic supplement were required to inhibit the growth of other intestinal bacteria. In this research, for sensitivity testing of Campylobacter jejuni and Campylobacter coli three different methods were used: disc diffusion test, E-test, and dilution agar method. A total of 55 strains of Campylobacter jejuni and Campylobacter coli. Out of the total, 24 strains originated from man, 16 from broilers were isolated, and 15 from pigs. Multidrug resistance was determined in cases when the strains were resistant to two or more antibiotics. Applying E-test, we detected that the largest number of Campylobacter jejuni were multi drug resistant to two antibiotics (41.2%, and three antibiotics (11.8%. Applying disc diffusion method it was detected that 5.9% of Campylobacter jejuni from man was resistant to four tested antibiotics. Applying all three methods, it was detected that the largest number of Campylobacter strains was resistant to two antibiotics and three antibiotics. Applying disc diffusion method it was detected that 50% of Campylobacter coli strains from pigs were resistant to three tested antibiotics.

  1. The Impact of Prophage on the Equilibria and Stability of Phage and Host

    Science.gov (United States)

    Yu, Pei; Nadeem, Alina; Wahl, Lindi M.

    2016-11-01

    In this paper, we present a bacteriophage model that includes prophage, that is, phage genomes that are incorporated into the host cell genome. The general model is described by an 18-dimensional system of ordinary differential equations. This study focuses on asymptotic behaviour of the model, and thus the system is reduced to a simple six-dimensional model, involving uninfected host cells, infected host cells and phage. We use dynamical system theory to explore the dynamic behaviour of the model, studying in particular the impact of prophage on the equilibria and stability of phage and host. We employ bifurcation and stability theory, centre manifold and normal form theory to show that the system has multiple equilibrium solutions which undergo a series of bifurcations, finally leading to oscillating motions. Numerical simulations are presented to illustrate and confirm the analytical predictions. The results of this study indicate that in some parameter regimes, the host cell population may drive the phage to extinction through diversification, that is, if multiple types of host emerge; this prediction holds even if the phage population is likewise diverse. This parameter regime is restricted, however, if infecting phage are able to recombine with prophage sequences in the host cell genome.

  2. The Impact of Prophage on the Equilibria and Stability of Phage and Host

    Science.gov (United States)

    Yu, Pei; Nadeem, Alina; Wahl, Lindi M.

    2017-06-01

    In this paper, we present a bacteriophage model that includes prophage, that is, phage genomes that are incorporated into the host cell genome. The general model is described by an 18-dimensional system of ordinary differential equations. This study focuses on asymptotic behaviour of the model, and thus the system is reduced to a simple six-dimensional model, involving uninfected host cells, infected host cells and phage. We use dynamical system theory to explore the dynamic behaviour of the model, studying in particular the impact of prophage on the equilibria and stability of phage and host. We employ bifurcation and stability theory, centre manifold and normal form theory to show that the system has multiple equilibrium solutions which undergo a series of bifurcations, finally leading to oscillating motions. Numerical simulations are presented to illustrate and confirm the analytical predictions. The results of this study indicate that in some parameter regimes, the host cell population may drive the phage to extinction through diversification, that is, if multiple types of host emerge; this prediction holds even if the phage population is likewise diverse. This parameter regime is restricted, however, if infecting phage are able to recombine with prophage sequences in the host cell genome.

  3. Factors Affecting Phage D29 Infection: A Tool to Investigate Different Growth States of Mycobacteria

    Science.gov (United States)

    Swift, Benjamin M. C.; Gerrard, Zara E.; Huxley, Jonathan N.; Rees, Catherine E. D.

    2014-01-01

    Bacteriophages D29 and TM4 are able to infect a wide range of mycobacteria, including pathogenic and non-pathogenic species. Successful phage infection of both fast- and slow-growing mycobacteria can be rapidly detected using the phage amplification assay. Using this method, the effect of oxygen limitation during culture of mycobacteria on the success of phage infection was studied. Both D29 and TM4 were able to infect cultures of M. smegmatis and Mycobacterium avium subspecies paratuberculosis (MAP) grown in liquid with aeration. However when cultures were grown under oxygen limiting conditions, only TM4 could productively infect the cells. Cell attachment assays showed that D29 could bind to the cells surface but did not complete the lytic cycle. The ability of D29 to productively infect the cells was rapidly recovered (within 1 day) when the cultures were returned to an aerobic environment and this recovery required de novo RNA synthesis. These results indicated that under oxygen limiting conditions the cells are entering a growth state which inhibits phage D29 replication, and this change in host cell biology which can be detected by using both phage D29 and TM4 in the phage amplification assay. PMID:25184428

  4. Factors affecting phage D29 infection: a tool to investigate different growth states of mycobacteria.

    Directory of Open Access Journals (Sweden)

    Benjamin M C Swift

    Full Text Available Bacteriophages D29 and TM4 are able to infect a wide range of mycobacteria, including pathogenic and non-pathogenic species. Successful phage infection of both fast- and slow-growing mycobacteria can be rapidly detected using the phage amplification assay. Using this method, the effect of oxygen limitation during culture of mycobacteria on the success of phage infection was studied. Both D29 and TM4 were able to infect cultures of M. smegmatis and Mycobacterium avium subspecies paratuberculosis (MAP grown in liquid with aeration. However when cultures were grown under oxygen limiting conditions, only TM4 could productively infect the cells. Cell attachment assays showed that D29 could bind to the cells surface but did not complete the lytic cycle. The ability of D29 to productively infect the cells was rapidly recovered (within 1 day when the cultures were returned to an aerobic environment and this recovery required de novo RNA synthesis. These results indicated that under oxygen limiting conditions the cells are entering a growth state which inhibits phage D29 replication, and this change in host cell biology which can be detected by using both phage D29 and TM4 in the phage amplification assay.

  5. Factors affecting phage D29 infection: a tool to investigate different growth states of mycobacteria.

    Science.gov (United States)

    Swift, Benjamin M C; Gerrard, Zara E; Huxley, Jonathan N; Rees, Catherine E D

    2014-01-01

    Bacteriophages D29 and TM4 are able to infect a wide range of mycobacteria, including pathogenic and non-pathogenic species. Successful phage infection of both fast- and slow-growing mycobacteria can be rapidly detected using the phage amplification assay. Using this method, the effect of oxygen limitation during culture of mycobacteria on the success of phage infection was studied. Both D29 and TM4 were able to infect cultures of M. smegmatis and Mycobacterium avium subspecies paratuberculosis (MAP) grown in liquid with aeration. However when cultures were grown under oxygen limiting conditions, only TM4 could productively infect the cells. Cell attachment assays showed that D29 could bind to the cells surface but did not complete the lytic cycle. The ability of D29 to productively infect the cells was rapidly recovered (within 1 day) when the cultures were returned to an aerobic environment and this recovery required de novo RNA synthesis. These results indicated that under oxygen limiting conditions the cells are entering a growth state which inhibits phage D29 replication, and this change in host cell biology which can be detected by using both phage D29 and TM4 in the phage amplification assay.

  6. Pentavalent single-domain antibodies reduce Campylobacter jejuni motility and colonization in chickens.

    Directory of Open Access Journals (Sweden)

    Ali Riazi

    Full Text Available Campylobacter jejuni is the leading cause of bacterial foodborne illness in the world, with symptoms ranging from acute diarrhea to severe neurological disorders. Contaminated poultry meat is a major source of C. jejuni infection, and therefore, strategies to reduce this organism in poultry, are expected to reduce the incidence of Campylobacter-associated diseases. We have investigated whether oral administration of C. jejuni-specific single-domain antibodies would reduce bacterial colonization levels in chickens. Llama single-domain antibodies specific for C. jejuni were isolated from a phage display library generated from the heavy chain IgG variable domain repertoire of a llama immunized with C. jejuni flagella. Two flagella-specific single-domain antibodies were pentamerized to yield high avidity antibodies capable of multivalent binding to the target antigen. When administered orally to C. jejuni-infected two-day old chicks, the pentabodies significantly reduced C. jejuni colonization in the ceca. In vitro, the motility of the bacteria was also reduced in the presence of the flagella-specific pentabodies, suggesting the mechanism of action is through either direct interference with flagellar motility or antibody-mediated aggregation. Fluorescent microscopy and Western blot analyses revealed specific binding of the anti-flagella pentabodies to the C. jejuni flagellin.

  7. Pentavalent single-domain antibodies reduce Campylobacter jejuni motility and colonization in chickens.

    Science.gov (United States)

    Riazi, Ali; Strong, Philippa C R; Coleman, Russell; Chen, Wangxue; Hirama, Tomoko; van Faassen, Henk; Henry, Matthew; Logan, Susan M; Szymanski, Christine M; Mackenzie, Roger; Ghahroudi, Mehdi Arbabi

    2013-01-01

    Campylobacter jejuni is the leading cause of bacterial foodborne illness in the world, with symptoms ranging from acute diarrhea to severe neurological disorders. Contaminated poultry meat is a major source of C. jejuni infection, and therefore, strategies to reduce this organism in poultry, are expected to reduce the incidence of Campylobacter-associated diseases. We have investigated whether oral administration of C. jejuni-specific single-domain antibodies would reduce bacterial colonization levels in chickens. Llama single-domain antibodies specific for C. jejuni were isolated from a phage display library generated from the heavy chain IgG variable domain repertoire of a llama immunized with C. jejuni flagella. Two flagella-specific single-domain antibodies were pentamerized to yield high avidity antibodies capable of multivalent binding to the target antigen. When administered orally to C. jejuni-infected two-day old chicks, the pentabodies significantly reduced C. jejuni colonization in the ceca. In vitro, the motility of the bacteria was also reduced in the presence of the flagella-specific pentabodies, suggesting the mechanism of action is through either direct interference with flagellar motility or antibody-mediated aggregation. Fluorescent microscopy and Western blot analyses revealed specific binding of the anti-flagella pentabodies to the C. jejuni flagellin.

  8. Cost-effectiveness of interventions to control Campylobacter in the New Zealand poultry meat food supply.

    Science.gov (United States)

    Lake, Robin J; Horn, Beverley J; Dunn, Alex H; Parris, Ruth; Green, F Terri; McNickle, Don C

    2013-07-01

    An analysis of the cost-effectiveness of interventions to control Campylobacter in the New Zealand poultry supply examined a series of interventions. Effectiveness was evaluated in terms of reduced health burden measured by disability-adjusted life years (DALYs). Costs of implementation were estimated from the value of cost elements, determined by discussions with industry. Benefits were estimated by changing the inputs to a poultry food chain quantitative risk model. Proportional reductions in the number of predicted Campylobacter infections were converted into reductions in the burden of disease measured in DALYs. Cost-effectiveness ratios were calculated for each intervention, as cost per DALY reduction and the ratios compared. The results suggest that the most cost-effective interventions (lowest ratios) are at the primary processing stage. Potential phage-based controls in broiler houses were also highly cost-effective. This study is limited by the ability to quantify costs of implementation and assumptions required to estimate health benefits, but it supports the implementation of interventions at the primary processing stage as providing the greatest quantum of benefit and lowest cost-effectiveness ratios.

  9. The Staphylococci Phages Family: An Overview

    Directory of Open Access Journals (Sweden)

    Laurence Van Melderen

    2012-11-01

    Full Text Available Due to their crucial role in pathogenesis and virulence, phages of Staphylococcus aureus have been extensively studied. Most of them encode and disseminate potent staphylococcal virulence factors. In addition, their movements contribute to the extraordinary versatility and adaptability of this prominent pathogen by improving genome plasticity. In addition to S. aureus, phages from coagulase-negative Staphylococci (CoNS are gaining increasing interest. Some of these species, such as S. epidermidis, cause nosocomial infections and are therefore problematic for public health. This review provides an overview of the staphylococcal phages family extended to CoNS phages. At the morphological level, all these phages characterized so far belong to the Caudovirales order and are mainly temperate Siphoviridae. At the molecular level, comparative genomics revealed an extensive mosaicism, with genes organized into functional modules that are frequently exchanged between phages. Evolutionary relationships within this family, as well as with other families, have been highlighted. All these aspects are of crucial importance for our understanding of evolution and emergence of pathogens among bacterial species such as Staphylococci.

  10. Staphylococcal pathogenicity island DNA packaging system involving cos-site packaging and phage-encoded HNH endonucleases.

    Science.gov (United States)

    Quiles-Puchalt, Nuria; Carpena, Nuria; Alonso, Juan C; Novick, Richard P; Marina, Alberto; Penadés, José R

    2014-04-22

    Staphylococcal pathogenicity islands (SaPIs) are the prototypical members of a widespread family of chromosomally located mobile genetic elements that contribute substantially to intra- and interspecies gene transfer, host adaptation, and virulence. The key feature of their mobility is the induction of SaPI excision and replication by certain helper phages and their efficient encapsidation into phage-like infectious particles. Most SaPIs use the headful packaging mechanism and encode small terminase subunit (TerS) homologs that recognize the SaPI-specific pac site and determine SaPI packaging specificity. Several of the known SaPIs do not encode a recognizable TerS homolog but are nevertheless packaged efficiently by helper phages and transferred at high frequencies. In this report, we have characterized one of the non-terS-coding SaPIs, SaPIbov5, and found that it uses two different, undescribed packaging strategies. SaPIbov5 is packaged in full-sized phage-like particles either by typical pac-type helper phages, or by cos-type phages--i.e., it has both pac and cos sites--a configuration that has not hitherto been described for any mobile element, phages included--and uses the two different phage-coded TerSs. To our knowledge, this is the first example of SaPI packaging by a cos phage, and in this, it resembles the P4 plasmid of Escherichia coli. Cos-site packaging in Staphylococcus aureus is additionally unique in that it requires the HNH nuclease, carried only by cos phages, in addition to the large terminase subunit, for cos-site cleavage and melting.

  11. Sequencing and Characterization of Pseudomonas aeruginosa phage JG004

    Directory of Open Access Journals (Sweden)

    Bunk Boyke

    2011-05-01

    Full Text Available Abstract Background Phages could be an important alternative to antibiotics, especially for treatment of multiresistant bacteria as e.g. Pseudomonas aeruginosa. For an effective use of bacteriophages as antimicrobial agents, it is important to understand phage biology but also genes of the bacterial host essential for phage infection. Results We isolated and characterized a lytic Pseudomonas aeruginosa phage, named JG004, and sequenced its genome. Phage JG004 is a lipopolysaccharide specific broad-host-range phage of the Myoviridae phage family. The genome of phage JG004 encodes twelve tRNAs and is highly related to the PAK-P1 phage genome. To investigate phage biology and phage-host interactions, we used transposon mutagenesis of the P. aeruginosa host and identified P. aeruginosa genes, which are essential for phage infection. Analysis of the respective P. aeruginosa mutants revealed several characteristics, such as host receptor and possible spermidine-dependance of phage JG004. Conclusions Whole genome sequencing of phage JG004 in combination with identification of P. aeruginosa host genes essential for infection, allowed insights into JG004 biology, revealed possible resistance mechanisms of the host bacterium such as mutations in LPS and spermidine biosynthesis and can also be used to characterize unknown gene products in P. aeruginosa.

  12. Risk factors for antibiotic resistance in Campylobacter spp. isolated from raw poultry meat in Switzerland

    Directory of Open Access Journals (Sweden)

    Danuser Jürg

    2003-12-01

    Full Text Available Abstract Background The world-wide increase of foodborne infections with antibiotic resistant pathogens is of growing concern and is designated by the World Health Organization as an emerging public health problem. Thermophilic Campylobacter have been recognised as a major cause of foodborne bacterial gastrointestinal human infections in Switzerland and in many other countries throughout the world. Poultry meat is the most common source for foodborne cases caused by Campylobacter. Because all classes of antibiotics recommended for treatment of human campylobacteriosis are also used in veterinary medicine, in view of food safety, the resistance status of Campylobacter isolated from poultry meat is of special interest. Methods Raw poultry meat samples were collected throughout Switzerland and Liechtenstein at retail level and examined for Campylobacter spp. One strain from each Campylobacter-positive sample was selected for susceptibility testing with the disc diffusion and the E-test method. Risk factors associated with resistance to the tested antibiotics were analysed by multiple logistic regression. Results In total, 91 Campylobacter spp. strains were isolated from 415 raw poultry meat samples. Fifty-one strains (59% were sensitive to all tested antibiotics. Nineteen strains (22% were resistant to a single, nine strains to two antibiotics, and eight strains showed at least three antibiotic resistances. Resistance was observed most frequently to ciprofloxacin (28.7%, tetracycline (12.6%, sulphonamide (11.8%, and ampicillin (10.3%. One multiple resistant strain exhibited resistance to five antibiotics including ciprofloxacin, tetracycline, and erythromycin. These are the most important antibiotics for treatment of human campylobacteriosis. A significant risk factor associated with multiple resistance in Campylobacter was foreign meat production compared to Swiss meat production (odds ratio = 5.7. Conclusion Compared to the situation in other

  13. Multiplex PCR Assay for Identifi cation and Differentiation of Campylobacter jejuni and Campylobacter coli Isolates.

    Science.gov (United States)

    Pavlova, Maria R; Dobreva, Elina G; Ivanova, Katucha I; Asseva, Galina D; Ivanov, Ivan N; Petrov, Peter K; Velev, Valeri R; Tomova, Ivelina I; Tiholova, Maida M; Kantardjiev, Todor V

    2016-01-01

    Campylobacter spp. are important causative agents of gastrointestinal infections in humans. The most frequently isolated strains of this bacterial genus are Campylobacter jejuni and Campylobacter coli. To date, genetic methods for bacterial identification have not been used in Bulgaria. We optimized the multiplex PSR assay to identify Campylobacter spp. and differentiate C. jejuni from C. coli in clinical isolates. We also compared this method with the routinely used biochemical methods. To identify Campylobacter spp. and discriminate C. coli from C. jejuni in clinical isolates using multiplex PCR assay. Between February 2014 and January 2015 we studied 93 stool samples taken from patients with diarrheal syndrome and identified 40 species of Campylobacter spp. in them. The clinical material was cultured in microaerophilic atmosphere, the isolated strains being biochemically diff erentiated (hydrolysis of sodium hippurate for C. jejuni, and hydrolysis of indoxyl acetate for C. coli). DNA was isolated from the strains using QiaAmp MiniKit (QIAGEN, Germany). Twenty strains were tested with multiplex PCR for the presence of these genes: cadF, characteristic for Campylobacter spp., hipO for C. jejuni and asp for C. coli. The biochemical tests identified 16 strains of C. jejuni, 3 strains of C. coli, and 1 strain of C. upsaliensis. After the multiplex PCR assay the capillary gel electrophoresis confirmed 16 strains of C. jejuni, 2 strains of C. coli and 2 strains of Campylobacter spp. - because of the presence of the gene cadF. C. jejuni has the gene hipO, and it is possible that this gene may not be expressed in the biochemical differentiation yielding a negative reaction as a result. In comparison, we can conclude that the genetic differentiation is a more accurate method than the biochemical tests. The multiplex PCR assay is a fast, accurate method for identifi cation of Campylobacter spp. which makes it quite necessary in the clinical diagnostic practice.

  14. Validation of the ANSR® for Campylobacter Method for Detection of Thermophilic Campylobacter spp. in Chicken Carcass Rinse and Turkey Sponge Samples.

    Science.gov (United States)

    Viator, Ryan; Alles, Susan; Le, Quynh-Nhi; Hosking, Edan; Biswas, Preetha; Zhang, Lei; Tolan, Jerry; Meister, Evan; Tovar, Eric; Pinkava, Lisa; Mozola, Mark; Rice, Jennifer; Chen, Yi; Odumeru, Joseph; Ziemer, Wayne

    2016-11-01

    A performance validation of the ANSR® for Campylobacter method was conducted in selected matrixes. This assay used selective nicking enzyme amplification technology to amplify target genes. Samples were enriched for 20 to 24 h and then lysed. The assay was completed within 50 min using real-time detection in a combination incubator/fluorescence detector and software. When 50 distinct strains of Campylobacter jejuni, C. lari, or C. coli were tested for inclusivity, all 50 strains produced positive results. In exclusivity testing, 31 strains of related organisms, including seven nontarget Campylobacter strains and other common species, were evaluated. All 31 species generated negative ANSR assay results, including the nontarget Campylobacter strains. The ANSR for Campylobacter method was compared to the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook reference method using naturally contaminated chicken carcass rinse or turkey carcass sponge samples. ANSR method performance was not statistically different from the reference method using two different enrichment options. Equivalent results were observed at both time points (20 and 24 h) and in both atmospheres (microaerobic and aerobic) to reference methods. Method performance with chicken carcass rinse was confirmed in an independent laboratory study. Additionally, in robustness testing, small, deliberate changes to the assay parameters minimally affected ANSR method performance. Finally, accelerated stability results from three independently manufactured lots supported a shelf life of 6 months when stored at 4°C. The ANSR assay offered greater efficiency and flexibility when compared to the reference method with a 20-24 h single-step enrichment in a microaerobic or an aerobic atmosphere.

  15. A Yersinia pestis-specific, lytic phage preparation significantly reduces viable Y. pestis on various hard surfaces experimentally contaminated with the bacterium

    OpenAIRE

    Rashid, Mohammed H.; Revazishvili, Tamara; Dean, Timothy; Butani, Amy; Verratti, Kathleen; Bishop-Lilly, Kimberly A.; Sozhamannan, Shanmuga; Sulakvelidze, Alexander; Rajanna, Chythanya

    2012-01-01

    Five Y. pestis bacteriophages obtained from various sources were characterized to determine their biological properties, including their taxonomic classification, host range and genomic diversity. Four of the phages (YpP-G, Y, R and YpsP-G) belong to the Podoviridae family, and the fifth phage (YpsP-PST) belongs to the Myoviridae family, of the order Caudovirales comprising of double-stranded DNA phages. The genomes of the four Podoviridae phages were fully sequenced and found to be almost id...

  16. Genetic heterogeneity of Campylobacter concisus determined by pulsed field gel electrophoresis-based macrorestriction profiling

    DEFF Research Database (Denmark)

    Matsheka, M.I.; Elisha, B.G.; Lastovica, A.L.

    2002-01-01

    In order to investigate the genetic diversity of Campylobacter concisus to assist molecular typing studies, the use of macrorestriction profiling was examined. A suitable protocol was developed that included the use of formaldehyde pretreatment to prevent DNA degradation, and restriction enzyme N...

  17. The complete genome sequences of 65 Campylobacter jejuni and C. coli strains

    Science.gov (United States)

    Campylobacter jejuni (Cj) and C. coli (Cc) are genetically highly diverse based on various molecular methods including MLST, microarray-based comparisons and the whole genome sequences of a few strains. Cj and Cc diversity is also exhibited by variable capsular polysaccharides (CPS) that are the maj...

  18. Energy Taxis Drives Campylobacter jejuni toward the Most Favorable Conditions for Growth

    DEFF Research Database (Denmark)

    Vegge, C.S.; Brondsted, L.; Li, Yiping

    2009-01-01

    Campylobacter jejuni is a serious food-borne bacterial pathogen in the developed world. Poultry is a major reservoir, and C. jejuni appears highly adapted to the gastrointestinal tract of birds. Several factors are important for chicken colonization and virulence, including a taxis mechanism...

  19. Farm specific risk factors for Campylobacter colonization of broilers in six European countries

    DEFF Research Database (Denmark)

    Borck Høg, Birgitte; Sommer, Helle Mølgaard; Williams, N.

    2015-01-01

    This study was part of the EU financed project CamCon. The objective of the study was to identify on-farm risk factors for Campylobacter colonization of broiler flocks based on comparable data from six European countries. The data included explanatory variables from a large questionnaire concerni...

  20. Analysis of farm specific risk factors for Campylobacter colonization of broilers in six European countries

    DEFF Research Database (Denmark)

    Sommer, Helle Mølgaard; Borck Høg, Birgitte; Larsen, Lars Stehr

    2016-01-01

    This study presents on-farm risk factors for the colonization of broiler flocks with Campylobacter based on comparable data from six European countries: Denmark, the Netherlands, Norway, Poland, Spain, and the UK. The study includes explanatory variables from a large questionnaire concerning prod...

  1. Sponge and skin excision sampling for recovery of Salmonella and Campylobacter from defeathered broiler carcasses

    Science.gov (United States)

    Introduction: Salmonella and Campylobacter contamination of broiler carcass skin increases during feather removal. There are several methods for sampling carcasses including sponging or swabbing of skin surface and skin excision. It is unclear whether sponge sampling is adequate to remove bacteria f...

  2. Occurrence, diversity, and host association of intestinal Campylobacter, Arcobacter, and Helicobacter in reptiles

    NARCIS (Netherlands)

    Gilbert, Maarten J; Kik, Marja; Timmerman, Arjen J; Severs, Tim T; Kusters, Johannes G; Duim, Birgitta; Wagenaar, Jaap A

    2014-01-01

    Campylobacter, Arcobacter, and Helicobacter species have been isolated from many vertebrate hosts, including birds, mammals, and reptiles. Multiple studies have focused on the prevalence of these Epsilonproteobacteria genera in avian and mammalian species. However, little focus has been given to the

  3. Characterization and lytic activity of Pseudomonas fluorescens phages from sewage

    Directory of Open Access Journals (Sweden)

    Ananthi Radhakrishnan

    2012-03-01

    Full Text Available Pseudomonas fluorescens phages from sewage were tested against P. fluorescens isolates of soil and sewage. The phages were characterized as to host range, morphology, structural proteins and genome fingerprint. Of the seven phages isolated, one was found to be abundant in sewage (5.9×10(7 pfu/mL, having broad host range, and distinct protein and DNA profile when compared to the other six phages. DNA restriction and protein profiles of the phages and their morphology indicate the diversity in the sewage environment. None of the isolates from the rhizosphere regions of various cultivated soils were susceptible to phages isolated from sewage.

  4. Application of Bacteroides fragilis phage as an alternative indicator of sewage pollution in Tampa Bay, Florida

    Science.gov (United States)

    McLaughlin, M.R.; Rose, J.B.

    2006-01-01

    Traditional fecal coliform bacterial indicators have been found to be severely limited in determining the significance and sources of fecal contamination in ambient waters of tropical and subtropical regions. The bacteriophages that infect Bacteroides fragilis have been suggested as better fecal indicators and at least one type may be human specific. In this study, the phages that infect B. fragilis host RYC2056 (RYC), including phage B56-3, and host ATCC 51477-HSP40 (HSP), including the human specific phage B40-8, were evaluated in the drainage basins of Tampa Bay, 7 samples (n = 62), or 11%, tested positive for the presence of phages infecting the host HSP, whereas 28 samples, or 45%, tested positive using the host RYC. A survival study was also done to compare the persistence of phages B56-3 and B40-8 to MS2 coliphage in seawater at various temperatures. The decay rates for MS2 were 0.239 log 10 d-1 at 10??C, but increased to 0.896 at 20??C and 2.62 log10 d-1 at 30??C. The two B. fragilis phages persisted much longer in the seawater compared to the coliphage and showed little variation between the temperatures. All sewage influents sampled from area wastewater treatment plants contained phages that infected the two B. fragilis hosts at levels from 1.2 ?? 104 to 1.11 ?? 10 5 pfu 100 ml-1 for host RYC and 67 to 350 pfu 100 ml -1 for host HSP. Of the 7 chlorinated effluent samples tested, 3 were positive for the presence of the phage using the host RYC and the phage enrichment method, with levels estimated to be <10 pfu 100 ml-1. No phages were detected using the host HSP in the treated sewage effluent. Coliphages were found in 3 of the 7 effluent samples at a range of 30 to 1.2 ?? 103 pfu 100 ml-1. ?? 2006 Estuarine Research Federation.

  5. Campylobacter

    Science.gov (United States)

    ... weakened from medications or other illnesses. Azithromycin and fluoroquinolones (e.g., ciprofloxacin) are commonly used for treatment of these infections, but resistance to fluoroquinolones is common. Antimicrobial susceptibility testing can help guide ...

  6. Burkholderia cepacia complex Phage-Antibiotic Synergy (PAS): antibiotics stimulate lytic phage activity.

    Science.gov (United States)

    Kamal, Fatima; Dennis, Jonathan J

    2015-02-01

    The Burkholderia cepacia complex (Bcc) is a group of at least 18 species of Gram-negative opportunistic pathogens that can cause chronic lung infection in cystic fibrosis (CF) patients. Bcc organisms possess high levels of innate antimicrobial resistance, and alternative therapeutic strategies are urgently needed. One proposed alternative treatment is phage therapy, the therapeutic application of bacterial viruses (or bacteriophages). Recently, some phages have been observed to form larger plaques in the presence of sublethal concentrations of certain antibiotics; this effect has been termed phage-antibiotic synergy (PAS). Those reports suggest that some antibiotics stimulate increased production of phages under certain conditions. The aim of this study is to examine PAS in phages that infect Burkholderia cenocepacia strains C6433 and K56-2. Bcc phages KS12 and KS14 were tested for PAS, using 6 antibiotics representing 4 different drug classes. Of the antibiotics tested, the most pronounced effects were observed for meropenem, ciprofloxacin, and tetracycline. When grown with subinhibitory concentrations of these three antibiotics, cells developed a chain-like arrangement, an elongated morphology, and a clustered arrangement, respectively. When treated with progressively higher antibiotic concentrations, both the sizes of plaques and phage titers increased, up to a maximum. B. cenocepacia K56-2-infected Galleria mellonella larvae treated with phage KS12 and low-dose meropenem demonstrated increased survival over controls treated with KS12 or antibiotic alone. These results suggest that antibiotics can be combined with phages to stimulate increased phage production and/or activity and thus improve the efficacy of bacterial killing.

  7. The Study of Campylobacter Frequency in Foodborne Disease Outbreaks in Iran

    Directory of Open Access Journals (Sweden)

    Mohammad Mehdi Soltan Dallal

    2017-07-01

    Full Text Available Introduction:Food-borne diseases are the widespread problems worldwide. Campylobacter species are the most important pathogens causing gastroenteritis which is generally transmitted through the food with animal origin. The aim of this study was to identify the Campylobacter spp. in diarrheal outbreaks in food-borne diseases at country level. Methods:This cross-sectional study carried out in spring and summer, 2015. In total, 305 swabs from diarrheal stool samples of 102 food-borne outbreaks were collected in various provinces of Iran. All samples were examined for the presence and growth of Campylobacter spp. The descriptive analysis, chi-square test and SPSS v.21 software were used for the analysis of results. Results:From the total of 102 foodborne outbreaks, Zanjan Province with 24 outbreaks (24.5% and Yazd Province with 70 samples of diarrhea (23% included the most reported cases. Out of 305 tested samples, 119 (39% were from food, 35 (11.5% from water and 151 (41.5% from unknown sources (p <0.001.  Two outbreaks in Yazd Province including three stool samples contained Campylobacter coli. Typical symptoms included diarrhea (30.9%, abdominal cramps (68.5%, fever (31.8%, headache (42.3%, diarrhea (5.2%, nausea (62.3% and vomiting (64.9%. Conclusion:Results from this study showed that C. coli was responsible for diarrhea rather than C. jejuni.

  8. Survival at refrigeration and freezing temperatures of Campylobacter coli and Campylobacter jejuni on chicken skin applied as axenic and mixed inoculums.

    Science.gov (United States)

    El-Shibiny, Ayman; Connerton, Phillippa; Connerton, Ian

    2009-05-31

    Campylobacter is considered to be the most common cause of bacterial diarrhoeal illness in the developed world. Many cases are thought to be acquired from consumption of undercooked poultry. The aim of this study was to compare the effect of the rate of cooling on the survival, at 4 degrees C and -20 degrees C, of Campylobacter coli and Campylobacter jejuni strains, inoculated on chicken skin from axenic culture or as mixed inoculums. Strains chilled in a domestic refrigerator varied in their tolerance to storage at 4 degrees C. Statistically significant differences between strains applied as axenic or mixed inoculums were observed for specific strain combinations using two-way ANOVA, including the enhanced survival of antibiotic resistant C. coli 99/367 at 4 degrees C. The use of rapid cooling (at -20 degrees C/min) enhanced the survival of all the Campylobacter strains chilled to 4 degrees C compared to standard refrigeration. Freezing to -20 degrees C reduced viable counts by 2.2-2.6 log10 CFU/cm(2) in 24 h. Rapid cooling to -20 degrees C (at -30 degrees C/min) enhanced the survival of C. coli 99/367 compared to freezing in a domestic freezer. Statistically significant interaction terms between specific strains were observed in mixed inoculums chilled to -20 degrees C by freezing in a domestic freezer and by rapid chilling to -20 degrees C. Rapid chilling of poultry, particularly for 4 degrees C storage may enhance survival of Campylobacter and although this is an issue that affects meat quality, it should be considered by poultry processors.

  9. Foodborne Campylobacter: Infections, Metabolism, Pathogenesis and Reservoirs

    Directory of Open Access Journals (Sweden)

    Sharon V. R. Epps

    2013-11-01

    Full Text Available Campylobacter species are a leading cause of bacterial-derived foodborne illnesses worldwide. The emergence of this bacterial group as a significant causative agent of human disease and their propensity to carry antibiotic resistance elements that allows them to resist antibacterial therapy make them a serious public health threat. Campylobacter jejuni and Campylobacter coli are considered to be the most important enteropathogens of this genus and their ability to colonize and survive in a wide variety of animal species and habitats make them extremely difficult to control. This article reviews the historical and emerging importance of this bacterial group and addresses aspects of the human infections they cause, their metabolism and pathogenesis, and their natural reservoirs in order to address the need for appropriate food safety regulations and interventions.

  10. Foodborne Campylobacter: Infections, Metabolism, Pathogenesis and Reservoirs

    Science.gov (United States)

    Epps, Sharon V. R.; Harvey, Roger B.; Hume, Michael E.; Phillips, Timothy D.; Anderson, Robin C.; Nisbet, David J.

    2013-01-01

    Campylobacter species are a leading cause of bacterial-derived foodborne illnesses worldwide. The emergence of this bacterial group as a significant causative agent of human disease and their propensity to carry antibiotic resistance elements that allows them to resist antibacterial therapy make them a serious public health threat. Campylobacter jejuni and Campylobacter coli are considered to be the most important enteropathogens of this genus and their ability to colonize and survive in a wide variety of animal species and habitats make them extremely difficult to control. This article reviews the historical and emerging importance of this bacterial group and addresses aspects of the human infections they cause, their metabolism and pathogenesis, and their natural reservoirs in order to address the need for appropriate food safety regulations and interventions. PMID:24287853

  11. Quantification of Growth of Campylobacter and Extended Spectrum β-Lactamase Producing Bacteria Sheds Light on Black Box of Enrichment Procedures.

    Science.gov (United States)

    Hazeleger, Wilma C; Jacobs-Reitsma, Wilma F; den Besten, Heidy M W

    2016-01-01

    Campylobacter is well recognized as the leading cause of bacterial foodborne diarrheal disease worldwide, and is routinely found in meat originating from poultry, sheep, pigs, and cattle. Effective monitoring of Campylobacter contamination is dependent on the availability of reliable detection methods. The method of the International Organization for Standardization for the detection of Campylobacter spp. in food (ISO 10272-1:2006) recommends the use of Bolton broth (BB) as selective enrichment medium, including a pre-enrichment step of 4-6 h at 37°C to revive sublethally damaged cells prior to incubation for 2 days at 41.5°C. Recently the presence of abundantly growing extended spectrum β-lactamase producing Enterobacteriaceae (ESBL bacteria) has become one of the most important factors that interfere with the isolation of Campylobacter, resulting in false-negative detection. However, detailed growth dynamics of Campylobacter and its competitors remain unclear, where these would provide a solid base for further improvement of the enrichment procedure for Campylobacter. Other enrichment broths, such as Preston broth (PB) and BB plus clavulanic acid (BBc) have been suggested to inhibit competitive flora. Therefore, these different broths were used as enrichments to measure the growth kinetics of several strains of Campylobacter jejuni and ESBL bacteria separately, in co-culture and of strains in chicken samples. The maximum cell numbers and often the growth rates of Campylobacter in mixed culture with ESBL bacteria were significantly lower than in single cultures, indicating severe suppression of Campylobacter by ESBL bacteria, also in naturally contaminated samples. PB and BBc successfully diminished ESBL bacteria and might therefore be a better choice as enrichment medium in possibly ESBL-bacteria contaminated samples. The efficacy of a pre-enrichment step in the BB ISO-procedure was not supported for cold-stressed and non-stressed cells. Therefore, omission of

  12. Comparative genomics defines the core genome of the growing N4-like phage genus and identifies N4-like Roseophage specific genes

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    Jacqueline Zoe-Munn Chan

    2014-10-01

    Full Text Available Two bacteriophages, RPP1 and RLP1, infecting members of the marine Roseobacter clade were isolated from seawater. Their linear genomes are 74.7 and 74.6 kb and encode 91 and 92 coding DNA sequences, respectively. Around 30% of these are homologous to genes found in Enterobacter phage N4. Comparative genomics of these two new Roseobacter phages and twenty-three other sequenced N4-like phages (three infecting members of the Roseobacter lineage and twenty infecting other Gammaproteobacteria revealed that N4-like phages share a core genome of 14 genes responsible for control of gene expression, replication and virion proteins. Phylogenetic analysis of these genes placed the five N4-like roseophages (RN4 into a distinct subclade. Analysis of the RN4 phage genomes revealed they share a further 19 genes of which nine are found exclusively in RN4 phages and four appear to have been acquired from their bacterial hosts. Proteomic analysis of the RPP1 and RLP1 virions identified a second structural module present in the RN4 phages similar to that found in the Pseudomonas N4-like phage LIT1. Searches of various metagenomic databases, included the GOS database, using CDS sequences from RPP1 suggests these phages are widely distributed in marine environments in particular in the open ocean environment.

  13. Spontaneous deletion of a 209-kilobase-pair fragment from the Escherichia coli genome occurs with acquisition of resistance to an assortment of infectious phages.

    Science.gov (United States)

    Tanji, Yasunori; Hattori, Kenji; Suzuki, Kohichi; Miyanaga, Kazuhiko

    2008-07-01

    To breed resistance to an assortment of infectious phages, continuous cultures of Escherichia coli JM109 grown in a chemostat were exposed to phage mixtures prepared from sewage influent. Four sequential chemostat-grown cultures were each infected with a different phage mixture. At the end of a chemostat run, one phage-resistant colony was isolated and used to inoculate the subsequent culture. This process was repeated, and increased phage resistance of the input bacterial strain resulted from the successive challenges with different phage cocktails. Multiple mutations apparently accumulated progressively. A mutant isolated at the end of the four runs, designated D198, showed resistance to 38 of 40 phages that infect the parent strain, JM109. D198 produced less outer membrane protein C (OmpC) than JM109. However, restoration of the OmpC protein by plasmid-mediated complementation did not completely restore the susceptibility of D198 to the 38 phages. Therefore, alterations beyond the level of OmpC protein production contribute to the phage resistance of D198. PCR-based genetic analysis revealed that D198 has a genome that is 209 kbp (about 200 genes) smaller than JM109. The deletion includes the chromosomal section from ompC to wbbL that encodes the rhamnosyl transferase involved in lipopolysaccharide biosynthesis. Strains D198 and JM109 were comparable in their growth characteristics and their abilities to express a recombinant protein.

  14. Invasion Assays and Genomotyping to Investigate Differences in Virulence of Campylobacter spp. Isolates from Iceland

    Science.gov (United States)

    Campylobacter spp. are the leading cause of human gastroenteritis worldwide. Epithelial cell invasion is thought to be essential for Campylobacter spp. infection. Previous invasion studies with intestinal epithelial cells revealed that the ability of different Campylobacter jejuni isolates to inva...

  15. In Vivo Imaging of Molecularly Targeted Phage

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    Kimberly A. Kelly

    2006-12-01

    Full Text Available Rapid identification of in vivo affinity ligands would have far-reaching applications for imaging specific molecular targets, in vivo systems imaging, and medical use. We have developed a high-throughput method for identifying and optimizing ligands to map and image biologic targets of interest in vivo. We directly labeled viable phage clones with far-red fluorochromes and comparatively imaged them in vivo by multichannel fluorescence ratio imaging. Using Secreted Protein Acidic and Rich in Cysteine (osteonectin and vascular cell adhesion molecule-1 as model targets, we show that: 1 fluorescently labeled phage retains target specificity on labeling; 2 in vivo distribution can be quantitated (detection thresholds of ~ 300 phage/mm3 tissue throughout the entire depth of the tumor using fluorescent tomographic imaging; and 3 fluorescently labeled phage itself can serve as a replenishable molecular imaging agent. The described method should find widespread application in the rapid in vivo discovery and validation of affinity ligands and, importantly, in the use of fluorochrome-labeled phage clones as in vivo imaging agents.

  16. Resistance to quinolones in Campylobacter jejuni and Campylobacter coli from Danish broilers at farm level

    DEFF Research Database (Denmark)

    Pedersen, Karl; Wedderkopp, A.

    2003-01-01

    Aims : To investigate the prevalence of quinolone resistance among Campylobacter jejuni and Camp. coli isolates from Danish poultry at the farm level, as well as for the whole country. Methods and Results : Data and isolates were collected from a national surveillance of Campylobacter in poultry......-resistant variant. Conclusions : Overall, quinolone resistance among Campylobacter isolates from Danish broilers was 7.5% in 1998 and 1999; it was higher among Camp. coli than Camp. jejuni . Genetic diversity among resistant isolates was lower than among susceptible isolates, and certain clones existed in both...... a resistant and a susceptible variant. Some resistant clones appeared to persist on the farms and were repeatedly isolated from poultry flocks. Significance and Impact of the Study : The study is important for the understanding of persistence and dynamics of Campylobacter in broiler houses. It also highlights...

  17. Healthy puppies and kittens as carriers of Campylobacter spp., with special reference to Campylobacter upsaliensis

    DEFF Research Database (Denmark)

    Hald, Birthe; Madsen, Mogens

    1997-01-01

    Living in a household with a dog or cat has previously been identified as a significant risk factor for acquiring campylobacteriosis, in particular, with reference to Campylobacter upsaliensis infection. In a cross-sectional study carried out in Denmark between August and December 1996, 72 healthy...... puppies and 42 healthy kittens, aged between 11 and 17 weeks, were sampled for fecal campylobacter shedding by culture of rectal swab specimens on blood-free agar base with cefoperazone at 32 mg/liter and amphotericin at 10 mg/liter and on blood-free agar base with cefoperazone at 8 mg/liter, teicoplanin...... for Campylobacter spp., with a species distribution of 76% C. jejuni, 5% C. coli, and 19% C. upsaliensis, Of the kittens examined, two (5%) excreted campylobacters; both strains were C. upsaliensis, None of the chicken samples examined were found to be positive for C. upsaliensis. We concluded that young puppies...

  18. Healthy puppies and kittens as carriers of Campylobacter spp., with special reference to Campylobacter upsaliensis

    DEFF Research Database (Denmark)

    Hald, Birthe; Madsen, Mogens

    1997-01-01

    Living in a household with a dog or cat has previously been identified as a significant risk factor for acquiring campylobacteriosis, in particular, with reference to Campylobacter upsaliensis infection. In a cross-sectional study carried out in Denmark between August and December 1996, 72 healthy...... puppies and 42 healthy kittens, aged between 11 and 17 weeks, were sampled for fecal campylobacter shedding by culture of rectal swab specimens on blood-free agar base with cefoperazone at 32 mg/liter and amphotericin at 10 mg/liter and on blood-free agar base with cefoperazone at 8 mg/liter, teicoplanin...... for Campylobacter spp., with a species distribution of 76% C. jejuni, 5% C. coli, and 19% C. upsaliensis, Of the kittens examined, two (5%) excreted campylobacters; both strains were C. upsaliensis, None of the chicken samples examined were found to be positive for C. upsaliensis. We concluded that young puppies...

  19. Campylobacter jejuni and Campylobacter coli in wild birds on Danish livestock farms

    DEFF Research Database (Denmark)

    Hald, Birthe; Skov, Marianne Nielsine; Nielsen, Eva Møller

    2016-01-01

    , fat score, gender, and migration range were not found to be associated with Campylobacter spp. carriage. A correlation was found between the prevalence (%) of C. jejuni in wild birds and the proportions (%) of C. jejuni in both manure on cattle farms (R-2 = 0.92) and poultry farms (R-2 = 0......Background: Reducing the occurrence of campylobacteriosis is a food safety issue of high priority, as in recent years it has been the most commonly reported zoonosis in the EU. Livestock farms are of particular interest, since cattle, swine and poultry are common reservoirs of Campylobacter spp....... The farm environment provides attractive foraging and breeding habitats for some bird species reported to carry thermophilic Campylobacter spp. We investigated the Campylobacter spp. carriage rates in 52 wild bird species present on 12 Danish farms, sampled during a winter and a summer season, in order...

  20. Prevalence of Thermotolerant Campylobacter spp. in Chicken Meat in Croatia and Multilocus Sequence Typing of a Small Subset of Campylobacter jejuni and Campylobacter coli Isolates

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    Andrea Humski

    2016-01-01

    Full Text Available In order to detect thermotolerant Campylobacter spp., 241 samples of fresh chicken meat, at retail in Croatia, were analysed according to a standard method, followed by biochemical test and molecular polymerase chain reaction/restriction enzyme analysis for exact species determination. Campylobacter spp. prevalence was 73.86 %. Campylobacter jejuni and Campylobacter coli were isolated from 53.53 and 15.35 % of the samples, respectively. In 4.98 % of isolates thermotolerant Campylobacter spp. were not determined. The multi locus sequence typing method was used to evaluate genetic diversity of eight Campylobacter jejuni and four Campylobacter coli isolates. To our knowledge, these results of genotyping provided the first data on the presence of sequence types (STs and clonal complexes (CCs of Campylobacter jejuni and C. coli isolates in Croatia. By applying the multilocus sequence typing, a new allele of tkt gene locus was discovered and marked tkt508. The C. jejuni ST 6182 and C. coli ST 6183 genotypes were described for the fi rst time, and all other identified genotypes were clustered in the previously described sequence types and clonal complexes. These findings provide useful information on the prevalence and epidemiology of Campylobacter jejuni and C. coli in Croatia.

  1. Campylobacter jejuni in commercial eggs

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    Belchiolina Beatriz Fonseca

    2014-01-01

    Full Text Available This study evaluated the ability of Campylobacter jejuni to penetrate through the pores of the shells of commercial eggs and colonize the interior of these eggs, which may become a risk factor for human infection. Furthermore, this study assessed the survival and viability of the bacteria in commercial eggs. The eggs were placed in contact with wood shavings infected with C. jejuni to check the passage of the bacteria. In parallel, the bacteria were inoculated directly into the air chamber to assess the viability in the egg yolk. To determine whether the albumen and egg fertility interferes with the entry and survival of bacteria, we used varying concentrations of albumen and SPF and commercial eggs. C. jejuni was recovered in SPF eggs (fertile after three hours in contact with contaminated wood shavings but not in infertile commercial eggs. The colonies isolated in the SPF eggs were identified by multiplex PCR and the similarity between strains verified by RAPD-PCR. The bacteria grew in different concentrations of albumen in commercial and SPF eggs. We did not find C. jejuni in commercial eggs inoculated directly into the air chamber, but the bacteria were viable during all periods tested in the wood shavings. This study shows that consumption of commercial eggs infected with C. jejuni does not represent a potential risk to human health.

  2. Campylobacter jejuni: exposure assessment and hazard characterization : growth, survival and infectivity of Campylobacter jejuni

    OpenAIRE

    Verhoeff-Bakkenes, L.

    2012-01-01

    Campylobacter jejuni, a small, curved or spirally shaped highly motile microorganism, is identified as a major cause of bacterial gastroenteritis throughout the world. Serious complications such as the Guillain-Barré syndrome and reactive arthritis might occasionally follow infection. In this thesis data were generated in the fields of hazard characterization and exposure assessment, which were used to identify risk factors for Campylobacter. Although C. jejuni is generally seen as an ...

  3. Current taxonomy of phages infecting lactic acid bacteria

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    Jennifer eMahony

    2014-01-01

    Full Text Available Phages infecting lactic acid bacteria have been the focus of significant research attention over the past three decades. Through the isolation and characterization of hundreds of phage isolates, it has been possible to classify phages of the dairy starter and adjunct bacteria Lactococus lactis, Streptococcus thermophilus, Leuconostoc spp. and Lactobacillus spp. Among these, phages of L. lactis have been most thoroughly scrutinized and serve as an excellent model system to address issues that arise when attempting taxonomic classification of phages infecting other LAB species. Here, we present an overview of the current taxonomy of phages infecting LAB genera of industrial significance, the methods employed in these taxonomic efforts and how these may be employed for the taxonomy of phages of currently underrepresented and emerging phage species.

  4. HostPhinder: A Phage Host Prediction Tool

    DEFF Research Database (Denmark)

    Villarroel, Julia; Kleinheinz, Kortine Annina; Jurtz, Vanessa Isabell

    2016-01-01

    The current dramatic increase of antibiotic resistant bacteria has revitalised the interest in bacteriophages as alternative antibacterial treatment. Meanwhile, the development of bioinformatics methods for analysing genomic data places high-throughput approaches for phage characterization within...... reach. Here, we present HostPhinder, a tool aimed at predicting the bacterial host of phages by examining the phage genome sequence. Using a reference database of 2196 phages with known hosts, HostPhinder predicts the host species of a query phage as the host of the most genomically similar reference...... phages. As a measure of genomic similarity the number of co-occurring k-mers (DNA sequences of length k) is used. Using an independent evaluation set, HostPhinder was able to correctly predict host genus and species for 81% and 74% of the phages respectively, giving predictions for more phages than BLAST...

  5. Quantifying potential sources of surface water contamination with Campylobacter jejuni and Campylobacter coli.

    Science.gov (United States)

    Mughini-Gras, Lapo; Penny, Christian; Ragimbeau, Catherine; Schets, Franciska M; Blaak, Hetty; Duim, Birgitta; Wagenaar, Jaap A; de Boer, Albert; Cauchie, Henry-Michel; Mossong, Joel; van Pelt, Wilfrid

    2016-09-15

    Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water contamination with Campylobacter is largely unknown. In the Netherlands, the massive poultry culling to control the 2003 avian influenza epidemic coincided with a 44-50% reduction in human campylobacteriosis cases in the culling areas, suggesting substantial environment-mediated spread of poultry-borne Campylobacter. We inferred the origin of surface water Campylobacter jejuni and Campylobacter coli strains in Luxembourg and the Netherlands, as defined by multilocus sequence typing, by comparison to strains from poultry, pigs, ruminants, and wild birds, using the asymmetric island model for source attribution. Most Luxembourgish water strains were attributed to wild birds (61.0%), followed by poultry (18.8%), ruminants (15.9%), and pigs (4.3%); whereas the Dutch water strains were mainly attributed to poultry (51.7%), wild birds (37.3%), ruminants (9.8%), and pigs (1.2%). Attributions varied over seasons and surface water types, and geographical variation in the relative contribution of poultry correlated with the magnitude of poultry production at either the national or provincial level, suggesting that environmental dissemination of Campylobacter from poultry farms and slaughterhouses can be substantial in poultry-rich regions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Genotypic and phenotypic properties of cattle-associated Campylobacter and their implications to public health in the USA.

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    Yasser M Sanad

    Full Text Available Since cattle are a major source of food and the cattle industry engages people from farms to processing plants and meat markets, it is conceivable that beef-products contaminated with Campylobacter spp. would pose a significant public health concern. To better understand the epidemiology of cattle-associated Campylobacter spp. in the USA, we characterized the prevalence, genotypic and phenotypic properties of these pathogens. Campylobacter were detected in 181 (19.2% out of 944 fecal samples. Specifically, 71 C. jejuni, 132 C. coli, and 10 other Campylobacter spp. were identified. The prevalence of Campylobacter varied regionally and was significantly (P<0.05 higher in fecal samples collected from the South (32.8% as compared to those from the North (14.8%, Midwest (15.83%, and East (12%. Pulsed Field Gel Electrophoresis (PFGE analysis showed that C. jejuni and C. coli isolates were genotypically diverse and certain genotypes were shared across two or more of the geographic locations. In addition, 13 new C. jejuni and two C. coli sequence types (STs were detected by Multi Locus Sequence Typing (MLST. C. jejuni associated with clinically human health important sequence type, ST-61 which was not previously reported in the USA, was identified in the present study. Most frequently observed clonal complexes (CC were CC ST-21, CC ST-42, and CC ST-61, which are also common in humans. Further, the cattle associated C. jejuni strains showed varying invasion and intracellular survival capacity; however, C. coli strains showed a lower invasion and intracellular survival potential compared to C. jejuni strains. Furthermore, many cattle associated Campylobacter isolates showed resistance to several antimicrobials including ciprofloxacin, erythromycin, and gentamicin. Taken together, our results highlight the importance of cattle as a potential reservoir for clinically important Campylobacter.

  7. European regulatory conundrum of phage therapy.

    Science.gov (United States)

    Verbeken, Gilbert; De Vos, Daniel; Vaneechoutte, Mario; Merabishvili, Maya; Zizi, Martin; Pirnay, Jean-Paul

    2007-10-01

    The treatment of infectious diseases with antibiotics is becoming increasingly challenging. Very few new antimicrobials are in the pharmaceutical industry pipeline. One of the potential alternatives for antibiotics is phage therapy. Major obstacles for the clinical application of bacteriophages are a false perception of viruses as 'enemies of life' and the lack of a specific frame for phage therapy in the current Medicinal Product Regulation. Short-term borderline solutions under the responsibility of a Medical Ethical Committee and/or under the umbrella of the Declaration of Helsinki are emerging. As a long-term solution, however, we suggest the creation of a specific section for phage therapy under the Advanced Therapy Medicinal Product Regulation.

  8. Phage therapy in the food industry.

    Science.gov (United States)

    Endersen, Lorraine; O'Mahony, Jim; Hill, Colin; Ross, R Paul; McAuliffe, Olivia; Coffey, Aidan

    2014-01-01

    Despite advances in modern technologies, the food industry is continuously challenged with the threat of microbial contamination. The overuse of antibiotics has further escalated this problem, resulting in the increasing emergence of antibiotic-resistant foodborne pathogens. Efforts to develop new methods for controlling microbial contamination in food and the food processing environment are extremely important. Accordingly, bacteriophages (phages) and their derivatives have emerged as novel, viable, and safe options for the prevention, treatment, and/or eradication of these contaminants in a range of foods and food processing environments. Whole phages, modified phages, and their derivatives are discussed in terms of current uses and future potential as antimicrobials in the traditional farm-to-fork context, encompassing areas such as primary production, postharvest processing, biosanitation, and biodetection. The review also presents some safety concerns to ensure safe and effective exploitation of bacteriophages in the future.

  9. Comparative genomics of Bacillus thuringiensis phage 0305φ8-36: defining patterns of descent in a novel ancient phage lineage

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    Serwer Philip

    2007-10-01

    Full Text Available Abstract Background The recently sequenced 218 kb genome of morphologically atypical Bacillus thuringiensis phage 0305φ8-36 exhibited only limited detectable homology to known bacteriophages. The only known relative of this phage is a string of phage-like genes called BtI1 in the chromosome of B. thuringiensis israelensis. The high degree of divergence and novelty of phage genomes pose challenges in how to describe the phage from its genomic sequences. Results Phage 0305φ8-36 and BtI1 are estimated to have diverged 2.0 – 2.5 billion years ago. Positionally biased Blast searches aligned 30 homologous structure or morphogenesis genes between 0305φ8-36 and BtI1 that have maintained the same gene order. Functional clustering of the genes helped identify additional gene functions. A conserved long tape measure gene indicates that a long tail is an evolutionarily stable property of this phage lineage. An unusual form of the tail chaperonin system split to two genes was characterized, as was a hyperplastic homologue of the T4gp27 hub gene. Within this region some segments were best described as encoding a conservative array of structure domains fused with a variable component of exchangeable domains. Other segments were best described as multigene units engaged in modular horizontal exchange. The non-structure genes of 0305φ8-36 appear to include the remnants of two replicative systems leading to the hypothesis that the genome plan was created by fusion of two ancestral viruses. The case for a member of the RNAi RNA-directed RNA polymerase family residing in 0305φ8-36 was strengthened by extending the hidden Markov model of this family. Finally, it was noted that prospective transcriptional promoters were distributed in a gradient of small to large transcripts starting from a fixed end of the genome. Conclusion Genomic organization at a level higher than individual gene sequence comparison can be analyzed to aid in understanding large phage

  10. Phage lysin LysK can be truncated to its CHAP domain and retain lytic activity against live antibiotic-resistant staphylococci.

    Science.gov (United States)

    Horgan, Marianne; O'Flynn, Gary; Garry, Jennifer; Cooney, Jakki; Coffey, Aidan; Fitzgerald, Gerald F; Ross, R Paul; McAuliffe, Olivia

    2009-02-01

    A truncated derivative of the phage endolysin LysK containing only the CHAP (cysteine- and histidine-dependent amidohydrolase/peptidase) domain exhibited lytic activity against live clinical staphylococcal isolates, including methicillin-resistant Staphylococcus aureus. This is the first known report of a truncated phage lysin which retains high lytic activity against live staphylococcal cells.

  11. A Systematic Review Characterizing On-Farm Sources of Campylobacter spp. for Broiler Chickens

    Science.gov (United States)

    Agunos, Agnes; Waddell, Lisa; Léger, David; Taboada, Eduardo

    2014-01-01

    Campylobacter and antimicrobial-resistant Campylobacter are frequently isolated from broiler chickens worldwide. In Canada, campylobacteriosis is the third leading cause of enteric disease and the regional emergence of ciprofloxacin-resistant Campylobacter in broiler chickens has raised a public health concern. This study aimed to identify, critically appraise, and synthesize literature on sources of Campylobacter in broilers at the farm level using systematic review methodology. Literature searches were conducted in January 2012 and included electronic searches in four bibliographic databases. Relevant studies in French or English (n = 95) conducted worldwide in any year and all study designs were included. Risk of Bias and GRADE criteria endorsed by the Cochrane collaboration was used to assess the internal validity of the study and overall confidence in the meta-analysis. The categories for on-farm sources were: broiler breeders/vertical transfer (number of studies = 32), animals (n = 57), humans (n = 26), environment (n = 54), and water (n = 63). Only three studies examined the antimicrobial resistance profiles of Campylobacter from these on-farm sources. Subgroups of data by source and outcome were analyzed using random effect meta-analysis. The highest risk for contaminating a new flock appears to be a contaminated barn environment due to insufficient cleaning and disinfection, insufficient downtime, and the presence of an adjacent broiler flock. Effective biosecurity enhancements from physical barriers to restricting human movement on the farm are recommended for consideration to enhance local on-farm food safety programs. Improved sampling procedures and standardized laboratory testing are needed for comparability across studies. Knowledge gaps that should be addressed include farm-level drug use and antimicrobial resistance information, further evaluation of the potential for vertical transfer, and improved genotyping methods to

  12. Comparative analysis of antimicrobial resistance and genetic diversity of Campylobacter from broilers slaughtered in Poland.

    Science.gov (United States)

    Wieczorek, Kinga; Denis, Edyta; Osek, Jacek

    2015-10-01

    In the current study, the relationship of Campylobacter jejuni and Campylobacter coli strains isolated at slaughter was investigated using comparative analysis of antimicrobial resistance (AMR), virulence gene (VG) and PFGE profiling. A total of 254 Campylobacter isolates from poultry caeca and corresponding carcasses, including 139 C. jejuni and 115 C. coli strains were tested. The most prevalent resistance profiles observed in C. jejuni were ciprofloxacin, nalidixic acid and tetracycline (46 out of 139, 33.1% isolates) as well as ciprofloxacin, nalidixic acid, tetracycline and streptomycin among C. coli strains (34 out of 115, 29.6%). Multi-resistance was found more frequently among C. coli than C. jejuni (PCampylobacter isolates tested. All Campylobacter strains were classified into 154 different PFGE types. Among them, 56 profiles (28 C. jejuni and 28 C. coli) were common for at least two isolates including 9 clusters covering from 4 to 9 strains. Campylobacter composite types generated by a combination of 154 PFGE types, 10 AMR profiles and 19 VG patterns divided 178 distinct types with 95% similarity. The majority of the composite profiles (76 for C. jejuni and 58 for C. coli; 75.3% in total) included only one bacterial isolate. Furthermore, 11 pairs of C. jejuni and 12 pairs of C. coli from caeca and the corresponding carcasses isolated from the same places possessed the identical PFGE, AMR and VG patterns. This study demonstrated that C. jejuni and C. coli isolated from poultry in Poland showed to have a high genetic diversity and a weak clonal population structure. However, the composite analysis revealed a strong evidence for cross-contamination of chicken carcasses during the slaughter process. Additionally, our results confirm that Campylobacter may easily contaminate poultry carcasses at slaughter process and spread around country. More than half of Campylobacter strains tested (50.4%) were resistant to at least two classes of antimicrobials, i

  13. A systematic review characterizing on-farm sources of Campylobacter spp. for broiler chickens.

    Directory of Open Access Journals (Sweden)

    Agnes Agunos

    Full Text Available Campylobacter and antimicrobial-resistant Campylobacter are frequently isolated from broiler chickens worldwide. In Canada, campylobacteriosis is the third leading cause of enteric disease and the regional emergence of ciprofloxacin-resistant Campylobacter in broiler chickens has raised a public health concern. This study aimed to identify, critically appraise, and synthesize literature on sources of Campylobacter in broilers at the farm level using systematic review methodology. Literature searches were conducted in January 2012 and included electronic searches in four bibliographic databases. Relevant studies in French or English (n = 95 conducted worldwide in any year and all study designs were included. Risk of Bias and GRADE criteria endorsed by the Cochrane collaboration was used to assess the internal validity of the study and overall confidence in the meta-analysis. The categories for on-farm sources were: broiler breeders/vertical transfer (number of studies = 32, animals (n = 57, humans (n = 26, environment (n = 54, and water (n = 63. Only three studies examined the antimicrobial resistance profiles of Campylobacter from these on-farm sources. Subgroups of data by source and outcome were analyzed using random effect meta-analysis. The highest risk for contaminating a new flock appears to be a contaminated barn environment due to insufficient cleaning and disinfection, insufficient downtime, and the presence of an adjacent broiler flock. Effective biosecurity enhancements from physical barriers to restricting human movement on the farm are recommended for consideration to enhance local on-farm food safety programs. Improved sampling procedures and standardized laboratory testing are needed for comparability across studies. Knowledge gaps that should be addressed include farm-level drug use and antimicrobial resistance information, further evaluation of the potential for vertical transfer, and improved genotyping

  14. The Caulobacter crescentus phage phiCbK: genomics of a canonical phage

    Directory of Open Access Journals (Sweden)

    Gill Jason J

    2012-10-01

    Full Text Available Abstract Background The bacterium Caulobacter crescentus is a popular model for the study of cell cycle regulation and senescence. The large prolate siphophage phiCbK has been an important tool in C. crescentus biology, and has been studied in its own right as a model for viral morphogenesis. Although a system of some interest, to date little genomic information is available on phiCbK or its relatives. Results Five novel phiCbK-like C. crescentus bacteriophages, CcrMagneto, CcrSwift, CcrKarma, CcrRogue and CcrColossus, were isolated from the environment. The genomes of phage phiCbK and these five environmental phage isolates were obtained by 454 pyrosequencing. The phiCbK-like phage genomes range in size from 205 kb encoding 318 proteins (phiCbK to 280 kb encoding 448 proteins (CcrColossus, and were found to contain nonpermuted terminal redundancies of 10 to 17 kb. A novel method of terminal ligation was developed to map genomic termini, which confirmed termini predicted by coverage analysis. This suggests that sequence coverage discontinuities may be useable as predictors of genomic termini in phage genomes. Genomic modules encoding virion morphogenesis, lysis and DNA replication proteins were identified. The phiCbK-like phages were also found to encode a number of intriguing proteins; all contain a clearly T7-like DNA polymerase, and five of the six encode a possible homolog of the C. crescentus cell cycle regulator GcrA, which may allow the phage to alter the host cell’s replicative state. The structural proteome of phage phiCbK was determined, identifying the portal, major and minor capsid proteins, the tail tape measure and possible tail fiber proteins. All six phage genomes are clearly related; phiCbK, CcrMagneto, CcrSwift, CcrKarma and CcrRogue form a group related at the DNA level, while CcrColossus is more diverged but retains significant similarity at the protein level. Conclusions Due to their lack of any apparent relationship to

  15. Vi I typing phage for generalized transduction of Salmonella typhi.

    Science.gov (United States)

    Cerquetti, M C; Hooke, A M

    1993-01-01

    Salmonella typhi Vi typing phages were used to transduce temperature-sensitive (Ts) mutants of Salmonella typhi. Antibiotic resistance and Ts+ markers were transduced at high frequency (> 10(-4) per virulent phage). Several markers were cotransduced by phage Vi I, suggesting that it may be useful for mapping studies of the S. typhi genome. PMID:8349572

  16. HostPhinder: A Phage Host Prediction Tool

    Directory of Open Access Journals (Sweden)

    Julia Villarroel

    2016-05-01

    Full Text Available The current dramatic increase of antibiotic resistant bacteria has revitalised the interest in bacteriophages as alternative antibacterial treatment. Meanwhile, the development of bioinformatics methods for analysing genomic data places high-throughput approaches for phage characterization within reach. Here, we present HostPhinder, a tool aimed at predicting the bacterial host of phages by examining the phage genome sequence. Using a reference database of 2196 phages with known hosts, HostPhinder predicts the host species of a query phage as the host of the most genomically similar reference phages. As a measure of genomic similarity the number of co-occurring k-mers (DNA sequences of length k is used. Using an independent evaluation set, HostPhinder was able to correctly predict host genus and species for 81% and 74% of the phages respectively, giving predictions for more phages than BLAST and significantly outperforming BLAST on phages for which both had predictions. HostPhinder predictions on phage draft genomes from the INTESTI phage cocktail corresponded well with the advertised targets of the cocktail. Our study indicates that for most phages genomic similarity correlates well with related bacterial hosts. HostPhinder is available as an interactive web service [1] and as a stand alone download from the Docker registry [2].

  17. Vi I typing phage for generalized transduction of Salmonella typhi.

    OpenAIRE

    Cerquetti, M C; Hooke, A M

    1993-01-01

    Salmonella typhi Vi typing phages were used to transduce temperature-sensitive (Ts) mutants of Salmonella typhi. Antibiotic resistance and Ts+ markers were transduced at high frequency (> 10(-4) per virulent phage). Several markers were cotransduced by phage Vi I, suggesting that it may be useful for mapping studies of the S. typhi genome.

  18. LuxS and quorum-sensing in Campylobacter

    Directory of Open Access Journals (Sweden)

    Paul ePlummer

    2012-03-01

    Full Text Available Several intercellular bacterial communication mechanisms have been identified in a broad range of bacterial species. These systems, collectively termed quorum-sensing systems, have been demonstrated to play significant roles in a variety of bacterial processes including motility, biofilm formation, expression of virulence genes and animal colonization. Campylobacter jejuni is known to poses a LuxS/ autoinducer-2 mediated system that has been partially characterized over the last decade. AI-2 is formed as a byproduct of the activated methyl recycling pathway, specifically by the LuxS enzyme. Previous work in our laboratory and that of others has demonstrated that this gene is involved in a variety of physiologic pathways of C. jejuni including motility, autoagglutination, CDT expression, flagellar expression, oxidative stress and animal colonization. This review article will summarize the current research associated with LuxS in C. jejuni and will provide insights into the role of this system in the metabolism and intercellular communication of this organism. Additionally, the evidence for other quorum sensing pathways in Campylobacter will be discussed.

  19. Cell biology perspectives in phage biology.

    Science.gov (United States)

    Ansaldi, Mireille

    2012-01-01

    Cellular biology has long been restricted to large cellular organisms. However, as the resolution of microscopic methods increased, it became possible to study smaller cells, in particular bacterial cells. Bacteriophage biology is one aspect of bacterial cell biology that has recently gained insight from cell biology. Despite their small size, bacteriophages could be successfully labeled and their cycle studied in the host cells. This review aims to put together, although non-extensively, several cell biology studies that recently pushed the elucidation of key mechanisms in phage biology, such as the lysis-lysogeny decision in temperate phages or genome replication and transcription, one step further.

  20. Dual role of Helicobacter and Campylobacter species in IBD: a systematic review and meta-analysis.

    Science.gov (United States)

    Castaño-Rodríguez, Natalia; Kaakoush, Nadeem O; Lee, Way Seah; Mitchell, Hazel M

    2017-02-01

    To conduct a comprehensive global systematic review and meta-analysis on the association between Helicobacter pylori infection and IBD. As bacterial antigen cross-reactivity has been postulated to be involved in this association, published data on enterohepatic Helicobacter spp (EHS) and Campylobacter spp and IBD was also analysed. Electronic databases were searched up to July 2015 for all case-control studies on H. pylori infection/EHS/Campylobacter spp and IBD. Pooled ORs (P-OR) and 95% CIs were obtained using the random effects model. Heterogeneity, sensitivity and stratified analyses were performed. Analyses comprising patients with Crohn's disease (CD), UC and IBD unclassified (IBDU), showed a consistent negative association between gastric H. pylori infection and IBD (P-OR: 0.43, p value Campylobacter spp, in particular C. concisus (P-OR: 3.76, p value=0.006) and C. showae (P-OR: 2.39, p value=0.027), increase IBD risk. H. pylori infection is negatively associated with IBD regardless of ethnicity, age, H. pylori detection methods and previous use of aminosalicylates and corticosteroids. Antibiotics influenced the magnitude of this association. Closely related bacteria including EHS and Campylobacter spp increase the risk of IBD. These results infer that H. pylori might exert an immunomodulatory effect in IBD. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  1. Changing risk of environmental Campylobacter exposure with emerging poultry production systems in Ethiopia.

    Science.gov (United States)

    Brena, M C; Mekonnen, Y; Bettridge, J M; Williams, N J; Wigley, P; Sisay Tessema, T; Christley, R M

    2016-02-01

    Campylobacter is a leading cause of diarrhoea, and its presence in chickens is a significant risk for zoonotic infection. Poultry production is becoming increasingly intensive in Ethiopia and is incorporating more high-producing breeds into traditionally managed smallholdings, especially in peri-urban areas. This cross-sectional study sampled 219 household environments in one peri-urban and two rural areas of Ethiopia, and an additional 20 semi-intensive farms in the peri-urban district. Campylobacter was detected by polymerase chain reaction (PCR)-specific assays in 44 samples; 16 of which could be identified as C. jejuni. Flocks in the peri-urban area were at significantly greater odds of detection, including those which only kept indigenous birds under a scavenging system. It was also noted that scavenging flocks of exotic high-production birds (Rhode Island Red) were at slightly greater risk, perhaps as exotic birds are under more stress when kept under traditional management systems. We suggest that changes to the system of chicken production may alter the ecology and epidemiology of Campylobacter in the environment, chickens and people, which may drive emergence of new epidemiological patterns of disease. Further research is needed to determine the extent to which the current management intensification and the distribution programmes of exotic and/or improved indigenous birds may alter Campylobacter epidemiology, ecology and public health risk, before their widespread adoption.

  2. Characterization of Antimicrobial Resistance Patterns and Detection of Virulence Genes in Campylobacter Isolates in Italy

    Science.gov (United States)

    Di Giannatale, Elisabetta; Di Serafino, Gabriella; Zilli, Katiuscia; Alessiani, Alessandra; Sacchini, Lorena; Garofolo, Giuliano; Aprea, Giuseppe; Marotta, Francesca

    2014-01-01

    Campylobacter has developed resistance to several antimicrobial agents over the years, including macrolides, quinolones and fluoroquinolones, becoming a significant public health hazard. A total of 145 strains derived from raw milk, chicken faeces, chicken carcasses, cattle faeces and human faeces collected from various Italian regions, were screened for antimicrobial susceptibility, molecular characterization (SmaI pulsed-field gel electrophoresis) and detection of virulence genes (sequencing and DNA microarray analysis). The prevalence of C. jejuni and C. coli was 62.75% and 37.24% respectively. Antimicrobial susceptibility revealed a high level of resistance for ciprofloxacin (62.76%), tetracycline (55.86%) and nalidixic acid (55.17%). Genotyping of Campylobacter isolates using PFGE revealed a total of 86 unique SmaI patterns. Virulence gene profiles were determined using a new microbial diagnostic microarray composed of 70-mer oligonucleotide probes targeting genes implicated in Campylobacter pathogenicity. Correspondence between PFGE and microarray clusters was observed. Comparisons of PFGE and virulence profiles reflected the high genetic diversity of the strains examined, leading us to speculate different degrees of pathogenicity inside Campylobacter populations. PMID:24556669

  3. Microscale electrodes integrated on COP for real sample Campylobacter spp. detection.

    Science.gov (United States)

    Morant-Miñana, M Carmen; Elizalde, J

    2015-08-15

    Campylobacter spp. are responsible for acute bacterial diseases in human worldwide. Nowadays campilobacteriosis is considered the most common foodborne illness in the European Union. In this paper the first electrochemical genosensor based on thin-film gold electrodes deposited onto Cyclo Olefin Polymer (COP) substrates was fabricated for the detection of Campylobacter spp in food matrices. The sensing element is characterized by several surface techniques and the sensitivity of the biosensor have been studied. A good linear relationship was obtained for the concentrations of PCR amplicon of Campylobacter spp. between 1 and 25 nM with a limit of detection (LOD) of 90 pM. Real samples have been validated with poultry meat samples and results were comparable with the PCR product samples. This is the last step for the fabrication of a Lab on a Chip (LOC), a biodevice integrating DNA sensor technology into microfluidic system, believed to perform an automated and complete assay, including sample preparation, PCR amplification, and electrochemical detection of Campylobacter spp. in raw poultry meat samples.

  4. Characterization of Antimicrobial Resistance Patterns and Detection of Virulence Genes in Campylobacter Isolates in Italy

    Directory of Open Access Journals (Sweden)

    Elisabetta Di Giannatale

    2014-02-01

    Full Text Available Campylobacter has developed resistance to several antimicrobial agents over the years, including macrolides, quinolones and fluoroquinolones, becoming a significant public health hazard. A total of 145 strains derived from raw milk, chicken faeces, chicken carcasses, cattle faeces and human faeces collected from various Italian regions, were screened for antimicrobial susceptibility, molecular characterization (SmaI pulsed-field gel electrophoresis and detection of virulence genes (sequencing and DNA microarray analysis. The prevalence of C. jejuni and C. coli was 62.75% and 37.24% respectively. Antimicrobial susceptibility revealed a high level of resistance for ciprofloxacin (62.76%, tetracycline (55.86% and nalidixic acid (55.17%. Genotyping of Campylobacter isolates using PFGE revealed a total of 86 unique SmaI patterns. Virulence gene profiles were determined using a new microbial diagnostic microarray composed of 70-mer oligonucleotide probes targeting genes implicated in Campylobacter pathogenicity. Correspondence between PFGE and microarray clusters was observed. Comparisons of PFGE and virulence profiles reflected the high genetic diversity of the strains examined, leading us to speculate different degrees of pathogenicity inside Campylobacter populations.

  5. Evaluating best practices for Campylobacter and Salmonella reduction in poultry processing plants.

    Science.gov (United States)

    Wideman, N; Bailey, M; Bilgili, S F; Thippareddi, H; Wang, L; Bratcher, C; Sanchez-Plata, M; Singh, M

    2016-02-01

    Poultry processing plants in the United States were surveyed on their current Campylobacter and Salmonella control practices. Following surveys, data were collected to develop a baseline for prevalence rates of Salmonella and Campylobacter; then changes in practices were implemented and evaluated for improvements in pathogen control. Surveys were sent to the plant Quality Assurance managers to determine production levels, antimicrobial interventions, and current pathogen testing practices. Initial sampling was performed at 6 plants with similar production volumes, at sites that included carcass samples before any pre-evisceration intervention, after exiting the inside-outside bird washer (IOBW), after exiting the pre-chiller, after exiting the primary chiller, and after exiting any post-chill intervention, as well as a water sample from each scalder, pre-chiller, primary chiller, and post-chill dip tank or finishing chiller. Enumerations and enrichments were performed for Campylobacter and Salmonella. Following the baseline sampling, changes in practices were suggested for each plant and a second sampling was conducted to determine their effectiveness. Results demonstrated that peracetic acid (PAA) was the most effective (P 0.05). Microbial buildup in the immersion tanks demonstrates the need for effective cleaning, sanitation practices, and chiller maintenance to reduce contamination of poultry with Campylobacter and Salmonella.

  6. Campylobacter polysaccharide capsules: virulence and vaccines

    Directory of Open Access Journals (Sweden)

    Patricia eGuerry

    2012-02-01

    Full Text Available Campylobacter jejuni remains a major cause of bacterial diarrhea worldwide and is associated with numerous sequelae, including Guillain Barre Syndrome, inflammatory bowel disease, reactive arthritis, and irritable bowel syndrome. C. jejuni is unusual for an intestinal pathogen in its ability to coat its surface with a polysaccharide capsule (CPS. These capsular polysaccharides vary in sugar composition and linkage, especially those involving heptoses of unusual configuration and O-methyl phosphoramidate linkages. This structural diversity is consistent with CPS being the major serodeterminant of the Penner scheme, of which there are 47 C. jejuni serotypes. Both CPS expression and expression of modifications are subject to phase variation by slip strand mismatch repair. Although capsules are virulence factors for other pathogens, the role of CPS in C. jejuni disease has not been well defined beyond descriptive studies demonstrating a role in serum resistance and for diarrhea in a ferret model of disease. However, perhaps the most compelling evidence for a role in pathogenesis are data that CPS conjugate vaccines protect against diarrheal disease in non-human primates. A CPS conjugate vaccine approach against this pathogen is intriguing, but several questions need to be addressed, including the valency of CPS types required for an effective vaccine. There have been numerous studies of prevalence of CPS serotypes in the developed world, but few studies from developing countries where the disease incidence is higher. The complexity and cost of Penner serotyping has limited its usefulness, and a recently developed multiplex PCR method for determination of capsule type offers the potential of a more rapid and affordable method. Comparative studies have shown a strong correlation of the two methods and studies are beginning to ascertain CPS-type distribution worldwide, as well as examination of correlation of severity of illness with specific CPS types.

  7. Thoracic empyema caused by Campylobacter rectus.

    Science.gov (United States)

    Ogata, Tomoyuki; Urata, Teruo; Nemoto, Daisuke; Hitomi, Shigemi

    2017-03-01

    We report a case of thoracic empyema caused by Campylobacter rectus, an organism considered as a periodontal pathogen but rarely recovered from extraoral specimens. The patient fully recovered through drainage of purulent pleural fluid and administration of antibiotics. The present case illustrates that C. rectus can be a cause of not only periodontal disease but also pulmonary infection.

  8. Campylobacter iguaniorum sp. nov., isolated from reptiles

    NARCIS (Netherlands)

    Gilbert, Maarten J.; Kik, Marja; Miller, William G.; Duim, Birgitta; Wagenaar, Jaap A.

    2015-01-01

    During sampling of reptiles for members of the class Epsilonproteobacteria, strains representing a member of the genus Campylobacter not belonging to any of the established taxa were isolated from lizards and chelonians. Initial amplified fragment length polymorphism, PCR and 16S rRNA sequence an

  9. Prevalence of Thermophilic Campylobacter species in carcasses ...

    African Journals Online (AJOL)

    Bernt Lindtjorn

    food animals and in bulk milk samples, tissue specimens ..... in Japan. J Clin Microbiol 2005; 43: 335–339. 8. Skirrow MB, Blaser MJ. Clinical ... Occurrence of. Campylobacter in retail foods in Ireland. Int J Food. Microbiol 2004; 95:111-118. 23.

  10. Studies on the epidemiology of Campylobacter jejuni

    NARCIS (Netherlands)

    J. Oosterom (Johannes)

    1985-01-01

    textabstractOver the last few years the bacterial species Campylobacter jejuni has been recognized as an important cause of acute enteritis in man. Investigations in several countries have shown that infections caused by C. jejuni may be as serious as those due to Salmonella spp., both in prevalence

  11. Diversity and geographical distribution of Flavobacterium psychrophilum isolates and their phages: patterns of susceptibility to phage infection and phage host range.

    Science.gov (United States)

    Castillo, Daniel; Christiansen, Rói Hammershaimb; Espejo, Romilio; Middelboe, Mathias

    2014-05-01

    Flavobacterium psychrophilum is an important fish pathogen worldwide that causes cold water disease (CWD) or rainbow trout fry syndrome (RTFS). Phage therapy has been suggested as an alternative method for the control of this pathogen in aquaculture. However, effective use of bacteriophages in disease control requires detailed knowledge about the diversity and dynamics of host susceptibility to phage infection. For this reason, we examined the genetic diversity of 49 F. psychrophilum strains isolated in three different areas (Chile, Denmark, and USA) through direct genome restriction enzyme analysis (DGREA) and their susceptibility to 33 bacteriophages isolated in Chile and Denmark, thus covering large geographical (>12,000 km) and temporal (>60 years) scales of isolation. An additional 40 phage-resistant isolates obtained from culture experiments after exposure to specific phages were examined for changes in phage susceptibility against the 33 phages. The F. psychrophilum and phage populations isolated from Chile and Denmark clustered into geographically distinct groups with respect to DGREA profile and host range, respectively. However, cross infection between Chilean phage isolates and Danish host isolates and vice versa was observed. Development of resistance to certain bacteriophages led to susceptibility to other phages suggesting that "enhanced infection" is potentially an important cost of resistance in F. psychrophilum, possibly contributing to the observed co-existence of phage-sensitive F. psychrophilum strains and lytic phages across local and global scales. Overall, our results showed that despite the identification of local communities of phages and hosts, some key properties determining phage infection patterns seem to be globally distributed.

  12. Complete genome sequence of Vibrio anguillarum phage CHOED successfully used for phage therapy in aquaculture

    DEFF Research Database (Denmark)

    Romero, Jaime; Higuera, Gastón; Gajardo, Felipe

    2014-01-01

    Vibrio anguillarum phage CHOED was isolated from Chilean mussels. It is a virulent phage showing effective inhibition of V. anguillarum. CHOED has potential in phage therapy, because it can protect fish from vibriosis in fish farms. Here, we announce the completely sequenced genome of V. anguilla...

  13. Occurrence, diversity, and host association of intestinal Campylobacter, Arcobacter, and Helicobacter in reptiles.

    Science.gov (United States)

    Gilbert, Maarten J; Kik, Marja; Timmerman, Arjen J; Severs, Tim T; Kusters, Johannes G; Duim, Birgitta; Wagenaar, Jaap A

    2014-01-01

    Campylobacter, Arcobacter, and Helicobacter species have been isolated from many vertebrate hosts, including birds, mammals, and reptiles. Multiple studies have focused on the prevalence of these Epsilonproteobacteria genera in avian and mammalian species. However, little focus has been given to the presence within reptiles, and their potential zoonotic and pathogenic roles. In this study, occurrence, diversity, and host association of intestinal Epsilonproteobacteria were determined for a large variety of reptiles. From 2011 to 2013, 444 cloacal swabs and fecal samples originating from 417 predominantly captive-held reptiles were screened for Epsilonproteobacteria. Campylobacter, Arcobacter, and Helicobacter genus specific PCRs were performed directly on all samples. All samples were also cultured on selective media and screened for the presence of Epsilonproteobacteria. Using a tiered approach of AFLP, atpA, and 16S rRNA sequencing, 432 Epsilonproteobacteria isolates were characterized at the species level. Based on PCR, Campylobacter, Arcobacter, and Helicobacter were detected in 69.3% of the reptiles; 82.5% of the chelonians, 63.8% of the lizards, and 58.0% of the snakes were positive for one or more of these genera. Epsilonproteobacteria were isolated from 22.1% of the reptiles and were isolated most frequently from chelonians (37.0%), followed by lizards (19.6%) and snakes (3.0%). The most commonly isolated taxa were Arcobacter butzleri, Arcobacter skirrowii, reptile-associated Campylobacter fetus subsp. testudinum, and a putative novel Campylobacter taxon. Furthermore, a clade of seven related putative novel Helicobacter taxa was isolated from lizards and chelonians. This study shows that reptiles carry various intestinal Epsilonproteobacteria taxa, including several putative novel taxa.

  14. Occurrence, Diversity, and Host Association of Intestinal Campylobacter, Arcobacter, and Helicobacter in Reptiles

    Science.gov (United States)

    Gilbert, Maarten J.; Kik, Marja; Timmerman, Arjen J.; Severs, Tim T.; Kusters, Johannes G.; Duim, Birgitta; Wagenaar, Jaap A.

    2014-01-01

    Campylobacter, Arcobacter, and Helicobacter species have been isolated from many vertebrate hosts, including birds, mammals, and reptiles. Multiple studies have focused on the prevalence of these Epsilonproteobacteria genera in avian and mammalian species. However, little focus has been given to the presence within reptiles, and their potential zoonotic and pathogenic roles. In this study, occurrence, diversity, and host association of intestinal Epsilonproteobacteria were determined for a large variety of reptiles. From 2011 to 2013, 444 cloacal swabs and fecal samples originating from 417 predominantly captive-held reptiles were screened for Epsilonproteobacteria. Campylobacter, Arcobacter, and Helicobacter genus specific PCRs were performed directly on all samples. All samples were also cultured on selective media and screened for the presence of Epsilonproteobacteria. Using a tiered approach of AFLP, atpA, and 16S rRNA sequencing, 432 Epsilonproteobacteria isolates were characterized at the species level. Based on PCR, Campylobacter, Arcobacter, and Helicobacter were detected in 69.3% of the reptiles; 82.5% of the chelonians, 63.8% of the lizards, and 58.0% of the snakes were positive for one or more of these genera. Epsilonproteobacteria were isolated from 22.1% of the reptiles and were isolated most frequently from chelonians (37.0%), followed by lizards (19.6%) and snakes (3.0%). The most commonly isolated taxa were Arcobacter butzleri, Arcobacter skirrowii, reptile-associated Campylobacter fetus subsp. testudinum, and a putative novel Campylobacter taxon. Furthermore, a clade of seven related putative novel Helicobacter taxa was isolated from lizards and chelonians. This study shows that reptiles carry various intestinal Epsilonproteobacteria taxa, including several putative novel taxa. PMID:24988130

  15. Occurrence, diversity, and host association of intestinal Campylobacter, Arcobacter, and Helicobacter in reptiles.

    Directory of Open Access Journals (Sweden)

    Maarten J Gilbert

    Full Text Available Campylobacter, Arcobacter, and Helicobacter species have been isolated from many vertebrate hosts, including birds, mammals, and reptiles. Multiple studies have focused on the prevalence of these Epsilonproteobacteria genera in avian and mammalian species. However, little focus has been given to the presence within reptiles, and their potential zoonotic and pathogenic roles. In this study, occurrence, diversity, and host association of intestinal Epsilonproteobacteria were determined for a large variety of reptiles. From 2011 to 2013, 444 cloacal swabs and fecal samples originating from 417 predominantly captive-held reptiles were screened for Epsilonproteobacteria. Campylobacter, Arcobacter, and Helicobacter genus specific PCRs were performed directly on all samples. All samples were also cultured on selective media and screened for the presence of Epsilonproteobacteria. Using a tiered approach of AFLP, atpA, and 16S rRNA sequencing, 432 Epsilonproteobacteria isolates were characterized at the species level. Based on PCR, Campylobacter, Arcobacter, and Helicobacter were detected in 69.3% of the reptiles; 82.5% of the chelonians, 63.8% of the lizards, and 58.0% of the snakes were positive for one or more of these genera. Epsilonproteobacteria were isolated from 22.1% of the reptiles and were isolated most frequently from chelonians (37.0%, followed by lizards (19.6% and snakes (3.0%. The most commonly isolated taxa were Arcobacter butzleri, Arcobacter skirrowii, reptile-associated Campylobacter fetus subsp. testudinum, and a putative novel Campylobacter taxon. Furthermore, a clade of seven related putative novel Helicobacter taxa was isolated from lizards and chelonians. This study shows that reptiles carry various intestinal Epsilonproteobacteria taxa, including several putative novel taxa.

  16. Targeting pancreatic islets with phage display assisted by laser pressure catapult microdissection.

    Science.gov (United States)

    Yao, Virginia J; Ozawa, Michael G; Trepel, Martin; Arap, Wadih; McDonald, Donald M; Pasqualini, Renata

    2005-02-01

    Heterogeneity of the microvasculature in different organs has been well documented by multiple methods including in vivo phage display. However, less is known about the diversity of blood vessels within functionally distinct regions of organs. Here, we combined in vivo phage display with laser pressure catapult microdissection to identify peptide ligands for vascular receptors in the islets of Langerhans in the murine pancreas. Protein database analyses of the peptides, CVSNPRWKC and CHVLWSTRC, showed sequence identity to two ephrin A-type ligand homologues, A2 and A4. Confocal microscopy confirmed that most immunoreactivity of CVSNPRWKC and CHVLWSTRC phage was associated with blood vessels in pancreatic islets. Antibodies recognizing EphA4, a receptor for ephrin-A ligands, were similarly associated with islet blood vessels. Importantly, binding of both islet-homing phage and anti-EphA4 antibody was strikingly increased in blood vessels of pancreatic islet tumors in RIP-Tag2 transgenic mice. These results indicate that endothelial cells of blood vessels in pancreatic islets preferentially express EphA4 receptors, and this expression is increased in tumors. Our findings show in vivo phage display and laser pressure catapult microdissection can be combined to reveal endothelial cell specialization within focal regions of the microvasculature.

  17. KSY1, a lactococcal phage with a T7-like transcription.

    Science.gov (United States)

    Chopin, Alain; Deveau, Hélène; Ehrlich, S Dusko; Moineau, Sylvain; Chopin, Marie-Christine

    2007-08-15

    The virulent lactococcal phage KSY1 possesses a large elongated capsid (223 nm long, 45 nm wide) and a short tail (32 nm). This phage of the Podoviridae group (C3 morphotype) has a linear 79,232-bp double-stranded DNA genome, which encodes 131 putative proteins and 3 tRNAs. This is the first description of the genome of a phage of this morphotype. KSY1 possesses a T7-like transcription system, including an RNA polymerase and a series of specific promoters, showing sequence homology to other known T7-like RNA polymerase promoters. Late stages of KSY1 multiplication are resistant to rifampicin. Otherwise, KSY1 shares limited similarity with other Podoviridae phages. Fourteen KSY1 structural proteins were identified by SDS-PAGE analysis. Among these proteins, those forming the distal tail structure and likely involved in host recognition are encoded by a 5-kb genomic region of KSY1. This region consists of a mosaic of DNA segments highly homologous to DNA of other lactococcal phages, suggesting an horizontal gene transfer.

  18. Genome-wide characterization of vibrio phage ϕpp2 with unique arrangements of the mob-like genes

    Directory of Open Access Journals (Sweden)

    Lin Ying-Rong

    2012-06-01

    Full Text Available Abstract Background Vibrio parahaemolyticus is associated with gastroenteritis, wound infections, and septicemia in human and animals. Phages can control the population of the pathogen. So far, the only one reported genome among giant vibriophages is KVP40: 244,835 bp with 26% coding regions that have T4 homologs. Putative homing endonucleases (HE were found in Vibrio phage KVP40 bearing one segD and Vibrio cholerae phage ICP1 carrying one mobC/E and one segG. Results A newly isolated Vibrio phage ϕpp2, which was specific to the hosts of V. parahaemolyticus and V. alginolyticus, featured a long nonenveloped head of ~90 × 150 nm and tail of ~110 nm. The phage can survive at 50°C for more than one hour. The genome of the phage ϕpp2 was sequenced to be 246,421 bp, which is 1587 bp larger than KVP40. 383 protein-encoding genes (PEGs and 30 tRNAs were found in the phage ϕpp2. Between the genomes of ϕpp2 and KVP40, 254 genes including 29 PEGs for viral structure were of high similarity, whereas 17 PEGs of KVP40 and 21 PEGs of ϕpp2 were unmatched. In both genomes, the capsid and tail genes have been identified, as well as the extensive representation of the DNA replication, recombination, and repair enzymes. In addition to the three giant indels of 1098, 1143 and 3330 nt, ϕpp2 possessed unique proteins involved in potassium channel, gp2 (DNA end protector, tRNA nucleotidyltransferase, and mob-type HEs, which were not reported in KVP40. The ϕpp2 PEG274, with strong promoters and translational initiation, was identified to be a mobE type, flanked by NrdA and NrdB/C homologs. Coincidently, several pairs of HE-flanking homologs with empty center were found in the phages of Vibrio phages ϕpp2 and KVP40, as well as in Aeromonas phages (Aeh1 and Ae65, and cyanophage P-SSM2. Conclusions Vibrio phage ϕpp2 was characterized by morphology, growth, and genomics with three giant indels and different types of HEs. The gene analysis on the required

  19. Phage display screen for peptides that bind Bcl-2 protein.

    Science.gov (United States)

    Park, Hye-Yeon; Kim, Joungmok; Cho, June-Haeng; Moon, Ji Young; Lee, Su-Jae; Yoon, Moon-Young

    2011-01-01

    Bcl-2 family proteins are key regulators of apoptosis associated with human disease, including cancer. Bcl-2 protein has been found to be overexpressed in many cancer cells. Therefore, Bcl-2 protein is a potential diagnostic target for cancer detection. In the present study, the authors have identified several Bcl-2 binding peptides with high affinity (picomolar range) from a 5-round M13 phage display library screening. These peptides can be used to develop novel diagnostic probes or potent inhibitors with diverse polyvalencies.

  20. Interactions between Phage-Shock Proteins in Escherichia coli

    OpenAIRE

    Adams, Hendrik; Teertstra, Wieke; Demmers, Jeroen; Boesten, Rolf; Tommassen, Jan

    2003-01-01

    Expression of the pspABCDE operon of Escherichia coli is induced upon infection by filamentous phage and by many other stress conditions, including defects in protein export. Expression of the operon requires the alternative sigma factor σ54 and the transcriptional activator PspF. In addition, PspA plays a negative regulatory role, and the integral-membrane proteins PspB and PspC play a positive one. In this study, we investigated whether the suggested protein-protein interactions implicated ...

  1. [Evaluation of usefulness of commercial recomwell Campylobacter enzyme--linked immunosorbent assays for routine serodiagnosis of Campylobacter jejuni and Campylobacter coli infections].

    Science.gov (United States)

    Rokosz, Natalia; Rastawicki, Waldemar; Jagielski, Marek

    2008-01-01

    The commercially available enzyme-linked immunosorbent assays (ELISA recomWell Campylobacter) from Mikrogen was evaluated for the diagnosis of Campylobacter jejuni and Campylobacter coli infections. Serum samples from 20 healthy controls, 44 persons with symptoms of primary Campylobacter infection and 24 serum samples from patients with Yersinia enterocolitica or Salmonella infections were tested. This ELISA assay detects IgA and IgG antibodies against three recombinant antigens of the Campylobacter jejuni and Campylobacter coli: OMP 18 (18 kDa), PEB4 (31 kDa) and P39 (39 kDa). The healthy controls showed significantly lower antibody titers in all two immunoglobulin classes. The IgA antibodies were diagnosed only in 2 (18.2%) serum samples obtained from patients with bacteriologically confirmed campylobacteriosis. The presence of IgG antibodies was confirmed in 82% of serum samples. Furthermore, we showed that 66.7% of the 33 serum samples obtained from the patients suspected for campylobacteriosis not confirmed by isolation, were positive for IgG and 15.2% for IgA antibodies. We observed also not specific reactions in ELISA recom Well Campylobacter with sera obtained form patients with yersiniosis and salmonelosis. This study demonstrates the usefulness of commercially available assay for the routine diagnosis of Campylobacter infection but with some limitations.

  2. Campylobacter pinnipediorum sp. nov., isolated from pinnipeds, comprising Campylobacter pinnipediorum subsp. pinnipediorum subsp. nov. and Campylobacter pinnipediorum subsp. caledonicus subsp. nov.

    Science.gov (United States)

    During independent diagnostic screenings of otariid seals in California (US) and phocid seals in Scotland (UK), Campylobacter-like isolates, which differed from the established Campylobacter taxa, were cultured from abscesses and internal organs of different seal species. A polyphasic study was unde...

  3. Does Campylobacter jejuni form biofilms in food-related environments?

    Science.gov (United States)

    Teh, Amy Huei Teen; Lee, Sui Mae; Dykes, Gary A

    2014-09-01

    Campylobacter jejuni is one of the most frequent causes of bacterial gastrointestinal food-borne infection worldwide. This species is part of the normal flora of the gastrointestinal tracts of animals used for food production, including poultry, which is regarded as the primary source of human Campylobacter infections. The survival and persistence of C. jejuni in food processing environments, especially in poultry processing plants, represent significant risk factors that contribute to the spread of this pathogen through the food chain. Compared to other food-borne pathogens, C. jejuni is more fastidious in its growth requirements and is very susceptible to various environmental stressors. Biofilm formation is suggested to play a significant role in the survival of C. jejuni in the food production and processing environment. The aims of this minireview were (i) to examine the evidence that C. jejuni forms biofilms and (ii) to establish the extent to which reported and largely laboratory-based studies of C. jejuni biofilms provide evidence for biofilm formation by this pathogen in food processing environments. Overall existing studies do not provide strong evidence for biofilm formation (as usually defined) by most C. jejuni strains in food-related environments under the combined conditions of atmosphere, temperature, and shear that they are likely to encounter. Simple attachment to and survival on surfaces and in existing biofilms of other species are far more likely to contribute to C. jejuni survival in food-related environments based on our current understanding of this species.

  4. Influence of environmental variables in the efficiency of phage therapy in aquaculture.

    Science.gov (United States)

    Silva, Yolanda J; Costa, Liliana; Pereira, Carla; Cunha, Ângela; Calado, Ricardo; Gomes, Newton C M; Almeida, Adelaide

    2014-09-01

    Aquaculture facilities worldwide continue to experience significant economic losses because of disease caused by pathogenic bacteria, including multidrug-resistant strains. This scenario drives the search for alternative methods to inactivate pathogenic bacteria. Phage therapy is currently considered as a viable alternative to antibiotics for inactivation of bacterial pathogens in aquaculture systems. While phage therapy appears to represent a useful and flexible tool for microbiological decontamination of aquaculture effluents, the effect of physical and chemical properties of culture waters on the efficiency of this technology has never been reported. The present study aimed to evaluate the effect of physical and chemical properties of aquaculture waters (e.g. pH, temperature, salinity and organic matter content) on the efficiency of phage therapy under controlled experimental conditions in order to provide a basis for the selection of the most suitable protocol for subsequent experiments. A bioluminescent genetically transformed Escherichia coli was selected as a model microorganism to monitor real-time phage therapy kinetics through the measurement of bioluminescence, thus avoiding the laborious and time-consuming conventional method of counting colony-forming units (CFU). For all experiments, a bacterial concentration of ≈ 10(5) CFU ml(-1) and a phage concentration of ≈ 10(6-8) plaque forming unit ml(-1) were used. Phage survival was not significantly affected by the natural variability of pH (6.5-7.4), temperature (10-25 °C), salinity (0-30 g NaCl l(-1) ) and organic matter concentration of aquaculture waters in a temperate climate. Nonetheless, the efficiency of phage therapy was mostly affected by the variation of salinity and organic matter content. As the effectiveness of phage therapy increases with water salt content, this approach appears to be a suitable choice for marine aquaculture systems. The success of phage therapy may also be enhanced in

  5. Phage & phosphatase: a novel phage-based probe for rapid, multi-platform detection of bacteria.

    Science.gov (United States)

    Alcaine, S D; Pacitto, D; Sela, D A; Nugen, S R

    2015-11-21

    Genetic engineering of bacteriophages allows for the development of rapid, highly specific, and easily manufactured probes for the detection of bacterial pathogens. A challenge for novel probes is the ease of their adoption in real world laboratories. We have engineered the bacteriophage T7, which targets Escherichia coli, to carry the alkaline phosphatase gene, phoA. This inclusion results in phoA overexpression following phage infection of E. coli. Alkaline phosphatase is commonly used in a wide range of diagnostics, and thus a signal produced by our phage-based probe could be detected using common laboratory equipment. Our work demonstrates the successful: (i) modification of T7 phage to carry phoA; (ii) overexpression of alkaline phosphatase in E. coli; and (iii) detection of this T7-induced alkaline phosphatase activity using commercially available colorimetric and chemilumiscent methods. Furthermore, we demonstrate the application of our phage-based probe to rapidly detect low levels of bacteria and discern the antibiotic resistance of E. coli isolates. Using our bioengineered phage-based probe we were able to detect 10(3) CFU per mL of E. coli in 6 hours using a chemiluminescent substrate and 10(4) CFU per mL within 7.5 hours using a colorimetric substrate. We also show the application of this phage-based probe for antibiotic resistance testing. We were able to determine whether an E. coli isolate was resistant to ampicillin within 4.5 hours using chemiluminescent substrate and within 6 hours using a colorimetric substrate. This phage-based scheme could be readily adopted in labs without significant capital investments and can be translated to other phage-bacteria pairs for further detection.

  6. Antimicrobial resistance in Campylobacter spp isolated from broiler flocks

    OpenAIRE

    Kuana, Suzete Lora; SANTOS Luciana Ruschel dos; RODRIGUES, Laura Beatriz; Anderlise BORSOI; Moraes, Hamilton Luis do Souza; Salle, Carlos Tadeu Pippi; Nascimento, Vladimir Pinheiro do

    2008-01-01

    The aim of this study was to assess the antimicrobial susceptibility of 62 Campylobacter spp. strains obtained from broiler flocks using the agar diffusion method. The Campylobacter spp strains were isolated from 22 flocks aged between 3 and 5 weeks of life, isolated from cloacae swabs, stools and cecal droppings in the farm and from the carcass rinsing in the slaughterhouse. Campylobacter spp strains were tested on Mueller-Hilton (MH) agar (27 samples) and MH plus TTC agar (35 samples). The ...

  7. Specific detection and confirmation of Campylobacter jejuni by DNA hybridization and PCR.

    Science.gov (United States)

    Ng, L K; Kingombe, C I; Yan, W; Taylor, D E; Hiratsuka, K; Malik, N; Garcia, M M

    1997-11-01

    Conventional detection and confirmation methods for Campylobacter jejuni are lengthy and tedious. A rapid hybridization protocol in which a 1,475-bp chromogen-labelled DNA probe (pDT1720) and Campylobacter strains filtered and grown on 0.22-micron-pore-size hydrophobic grid membrane filters (HGMFs) are used was developed. Among the environmental and clinical isolates of C. jejuni, Campylobacter coli, Campylobacter jejuni subsp. doylei, Campylobacter lari, and Arcobacter nitrofigilis and a panel of 310 unrelated bacterial strains tested, only C. jejuni and C. jejuni subsp. doylei isolates hybridized with the probe under stringent conditions. The specificity of the probe was confirmed when the protocol was applied to spiked skim milk and chicken rinse samples. Based on the nucleotide sequence of pDT1720, a pair of oligonucleotide primers was designed for PCR amplification of DNA from Campylobacter spp. and other food pathogens grown overnight in selective Mueller-Hinton broth with cefoperazone and growth supplements. All C. jejuni strains tested, including DNase-producing strains and C. jejuni subsp. doylei, produced a specific 402-bp amplicon, as confirmed by restriction and Southern blot analysis. The detection range of the assay was as low as 3 CFU per PCR to as high as 10(5) CFU per PCR for pure cultures. Overnight enrichment of chicken rinse samples spiked initially with as little as approximately 10 CFU/ml produced amplicons after the PCR. No amplicon was detected with any of the other bacterial strains tested or from the chicken background microflora. Since C. jejuni is responsible for 99% of Campylobacter contamination in poultry, PCR and HGMF hybridization were performed on naturally contaminated chicken rinse samples, and the results were compared with the results of conventional cultural isolation on Preston agar. All samples confirmed to be culture positive for C. jejuni were also identified by DNA hybridization and PCR amplification, thus confirming that

  8. Interaction Analysis through Proteomic Phage Display

    Directory of Open Access Journals (Sweden)

    Gustav N. Sundell

    2014-01-01

    Full Text Available Phage display is a powerful technique for profiling specificities of peptide binding domains. The method is suited for the identification of high-affinity ligands with inhibitor potential when using highly diverse combinatorial peptide phage libraries. Such experiments further provide consensus motifs for genome-wide scanning of ligands of potential biological relevance. A complementary but considerably less explored approach is to display expression products of genomic DNA, cDNA, open reading frames (ORFs, or oligonucleotide libraries designed to encode defined regions of a target proteome on phage particles. One of the main applications of such proteomic libraries has been the elucidation of antibody epitopes. This review is focused on the use of proteomic phage display to uncover protein-protein interactions of potential relevance for cellular function. The method is particularly suited for the discovery of interactions between peptide binding domains and their targets. We discuss the largely unexplored potential of this method in the discovery of domain-motif interactions of potential biological relevance.

  9. Recognition of epoxy with phage displayed peptides.

    Science.gov (United States)

    Swaminathan, Swathi; Cui, Yue

    2013-07-01

    The development of a general approach for non-destructive chemical and biological functionalization of epoxy could expand opportunities for both fundamental studies and creating various device platforms. Epoxy shows unique electrical, mechanical, chemical and biological compatibility and has been widely used for fabricating a variety of devices. Phage display has emerged as a powerful method for selecting peptides that possess enhanced selectivity and binding affinity toward a variety of targets. In this letter, we demonstrate for the first time a powerful yet benign approach for identifying binding motifs to epoxy via comprehensively screened phage displayed peptides. Our results show that the epoxy can be selectively recognized with peptide-displaying phages. Further, along with the development of epoxy-based microstructures; recognition of the epoxy with phage displayed peptides can be specifically localized in these microstructures. We anticipate that these results could open up exciting opportunities in the use of peptide-recognized epoxy in fundamental biochemical recognition studies, as well as in applications ranging from analytical devices, hybrid materials, surface and interface, to cell biology. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Phage-bacteria interaction network in human oral microbiome.

    Science.gov (United States)

    Wang, Jinfeng; Gao, Yuan; Zhao, Fangqing

    2016-07-01

    Although increasing knowledge suggests that bacteriophages play important roles in regulating microbial ecosystems, phage-bacteria interaction in human oral cavities remains less understood. Here we performed a metagenomic analysis to explore the composition and variation of oral dsDNA phage populations and potential phage-bacteria interaction. A total of 1,711 contigs assembled with more than 100 Gb shotgun sequencing data were annotated to 104 phages based on their best BLAST matches against the NR database. Bray-Curtis dissimilarities demonstrated that both phage and bacterial composition are highly diverse between periodontally healthy samples but show a trend towards homogenization in diseased gingivae samples. Significantly, according to the CRISPR arrays that record infection relationship between bacteria and phage, we found certain oral phages were able to invade other bacteria besides their putative bacterial hosts. These cross-infective phages were positively correlated with commensal bacteria while were negatively correlated with major periodontal pathogens, suggesting possible connection between these phages and microbial community structure in oral cavities. By characterizing phage-bacteria interaction as networks rather than exclusively pairwise predator-prey relationships, our study provides the first insight into the participation of cross-infective phages in forming human oral microbiota.

  11. The factors affecting effectiveness of treatment in phages therapy, mini review

    Directory of Open Access Journals (Sweden)

    Mai Huong eCHATAIN-LY

    2014-02-01

    Full Text Available In recent years, the use of lytic bacteriophages as antimicrobial agents controlling pathogenic bacteria has appeared as a promising new alternative strategy in the face of growing antibiotic resistance which has caused problems in many fields including medicine, veterinary medicine and aquaculture. The use of bacteriophages has numerous advantages over traditional antimicrobials. The effectiveness of phage applications in fighting against pathogenic bacteria depends on several factors such as the bacteriophages/target bacteria ratio, the mode and moment of treatment, environmental conditions (pH, temperature ..., the neutralization of phage and accessibility to target bacteria, amongst others. This report presents these factors and the challenges involved in developing phage therapy applications

  12. A Review of Phage Therapy against Bacterial Pathogens of Aquatic and Terrestrial Organisms

    Science.gov (United States)

    Doss, Janis; Culbertson, Kayla; Hahn, Delilah; Camacho, Joanna; Barekzi, Nazir

    2017-01-01

    Since the discovery of bacteriophage in the early 1900s, there have been numerous attempts to exploit their innate ability to kill bacteria. The purpose of this report is to review current findings and new developments in phage therapy with an emphasis on bacterial diseases of marine organisms, humans, and plants. The body of evidence includes data from studies investigating bacteriophage in marine and land environments as modern antimicrobial agents against harmful bacteria. The goal of this paper is to present an overview of the topic of phage therapy, the use of phage-derived protein therapy, and the hosts that bacteriophage are currently being used against, with an emphasis on the uses of bacteriophage against marine, human, animal and plant pathogens. PMID:28335451

  13. A Review of Phage Therapy against Bacterial Pathogens of Aquatic and Terrestrial Organisms

    Directory of Open Access Journals (Sweden)

    Janis Doss

    2017-03-01

    Full Text Available Since the discovery of bacteriophage in the early 1900s, there have been numerous attempts to exploit their innate ability to kill bacteria. The purpose of this report is to review current findings and new developments in phage therapy with an emphasis on bacterial diseases of marine organisms, humans, and plants. The body of evidence includes data from studies investigating bacteriophage in marine and land environments as modern antimicrobial agents against harmful bacteria. The goal of this paper is to present an overview of the topic of phage therapy, the use of phage-derived protein therapy, and the hosts that bacteriophage are currently being used against, with an emphasis on the uses of bacteriophage against marine, human, animal and plant pathogens.

  14. A Review of Phage Therapy against Bacterial Pathogens of Aquatic and Terrestrial Organisms.

    Science.gov (United States)

    Doss, Janis; Culbertson, Kayla; Hahn, Delilah; Camacho, Joanna; Barekzi, Nazir

    2017-03-18

    Since the discovery of bacteriophage in the early 1900s, there have been numerous attempts to exploit their innate ability to kill bacteria. The purpose of this report is to review current findings and new developments in phage therapy with an emphasis on bacterial diseases of marine organisms, humans, and plants. The body of evidence includes data from studies investigating bacteriophage in marine and land environments as modern antimicrobial agents against harmful bacteria. The goal of this paper is to present an overview of the topic of phage therapy, the use of phage-derived protein therapy, and the hosts that bacteriophage are currently being used against, with an emphasis on the uses of bacteriophage against marine, human, animal and plant pathogens.

  15. How to Name and Classify Your Phage: An Informal Guide

    Directory of Open Access Journals (Sweden)

    Evelien Adriaenssens

    2017-04-01

    Full Text Available With this informal guide, we try to assist both new and experienced phage researchers through two important stages that follow phage discovery; that is, naming and classification. Providing an appropriate name for a bacteriophage is not as trivial as it sounds, and the effects might be long-lasting in databases and in official taxon names. Phage classification is the responsibility of the Bacterial and Archaeal Viruses Subcommittee (BAVS of the International Committee on the Taxonomy of Viruses (ICTV. While the BAVS aims at providing a holistic approach to phage taxonomy, for individual researchers who have isolated and sequenced a new phage, this can be a little overwhelming. We are now providing these researchers with an informal guide to phage naming and classification, taking a “bottom-up” approach from the phage isolate level.

  16. Isolation of a Campylobacter lanienae-like bacterium from laboratory chinchillas (Chinchilla laniger).

    Science.gov (United States)

    Turowski, E E; Shen, Z; Ducore, R M; Parry, N M A; Kirega, A; Dewhirst, F E; Fox, J G

    2014-12-01

    Routine necropsies of 27 asymptomatic juvenile chinchillas revealed a high prevalence of gastric ulcers with microscopic lymphoplasmacytic gastroenteritis and typhlocolitis. Polymerase chain reaction (PCR) analysis using Campylobacter genus-specific partial 16S rRNA primers revealed the presence of Campylobacter spp. DNA in the faeces of 12 of 27 animals (44.4%). Species-specific partial 16S rRNA PCR and sequencing confirmed that these animals were colonized with Campylobacter lanienae, a gram-negative, microaerophilic bacterium that was first identified on routine faecal screening of slaughterhouse employees and subsequently isolated from faeces of livestock. Campylobacter lanienae was isolated from the faeces of six PCR-positive animals and identified with species-specific PCR and full 16S rRNA sequencing. Phylogenetic analysis showed that these isolates clustered with C. lanienae strain NCTC 13004. PCR analysis of DNA extracted from gastrointestinal tissues revealed the presence of C. lanienae DNA in the caecum and colon of these chinchillas. Gastrointestinal lesions were scored and compared between C. lanienae-positive and C. lanienae-negative animals. There was no correlation between colonization status and lesion severity in the stomach, liver, duodenum, or colon. Possible routes of C. lanienae infection in chinchillas could include waterborne transmission and faecal-oral transmission from wild mice and rats or livestock. Based on these findings, the authors conclude that C. lanienae colonizes the lower bowel of chinchillas in the absence of clinical disease. This is the first report of C. lanienae in any rodent species. Campylobacter lanienae isolates from different mammalian species demonstrate heterogeneity by 16S rRNA sequence comparison. Analysis using rpoB suggests that isolates and clones currently identified as C. lanienae may represent multiple species or subspecies.

  17. Campylobacter upsaliensis isolated from dogs produces high titer of cytolethal distending toxin

    Science.gov (United States)

    SOMROOP, Srinuan; HATANAKA, Noritoshi; AWASTHI, Sharda Prasad; OKUNO, Kentaro; ASAKURA, Masahiro; HINENOYA, Atsushi; YAMASAKI, Shinji

    2017-01-01

    Cytolethal distending toxin (CDT) consisting of CdtA, CdtB and CdtC has been reported to be a possible virulence factor of campylobacters including Campylobacter upsaliensis. In our previous study, the cdtB gene-based PCR-restriction fragment length polymorphism (RFLP) assay for detection and differentiation of 7 Campylobacter species yielded 3 different RFLP patterns (Cu-I to Cu-III). In this study, entire cdt (Cucdt) genes of each pattern were sequenced to see whether there are any differences in cdt genes, its amino acid sequences and biological activity of CuCDT. We found that all 3 representative strains harbor the entire Cucdt genes and homology between prototype and newly determined Cucdt genes was 94 to 98% with cdtA, 93 to 94% with cdtB and 92 to 93% with cdtC, while that between amino acids of CuCDT was 95 to 99% with CdtA, 97 to 98% with CdtB and 92 to 93% with CdtC. Furthermore, CDT activity produced by C. upsaliensis strains was examined by cytotoxicity assay with HeLa cells. Interestingly, C. upsaliensis produced 64 to 2,340 times higher CDT titer in comparison to other campylobacters did. In addition, Cu-III showed 64 times higher CDT titer than Cu-II, although CDT production level was almost the same by western blotting. These data suggest that CDT produced by C. upsaliensis might contribute more to human diseases in comparison to that produced by other campylobacters and Cu-III CDT seems to be more toxic to HeLa cells in comparison to Cu-I and Cu-II CDTs. PMID:28202878

  18. Thermophilic Campylobacter spp. in salad vegetables in Malaysia.

    Science.gov (United States)

    Chai, Lay Ching; Robin, Tunung; Ragavan, Usha Menon; Gunsalam, Jurin Wolmon; Bakar, Fatimah Abu; Ghazali, Farinazleen Mohamad; Radu, Son; Kumar, Malakar Pradeep

    2007-06-10

    The main aim of this study was to combine the techniques of most probable number (MPN) and polymerase chain reaction (PCR) for quantifying the prevalence and numbers of Campylobacter spp. in ulam, a popular Malaysian salad dish, from a traditional wet market and two modern supermarkets in Selangor, Malaysia. A total of 309 samples of raw vegetables which are used in ulam were examined in the study. The prevalences of campylobacters in raw vegetables were, for supermarket I, Campylobacter spp., 51.9%; Campylobacter jejuni, 40.7%; and Campylobacter coli, 35.2%: for supermarket II, Campylobacter spp., 67.7%; C. jejuni, 67.7%; and C. coli, 65.7%: and for the wet market, Campylobacter spp., 29.4%; C. jejuni, 25.5%; and C. coli, 22.6%. In addition Campylobacter fetus was detected in 1.9% of raw vegetables from supermarket I. The maximum numbers of Campylobacter spp. in raw vegetables from supermarkets and the wet market were >2400 and 460 MPN/g, respectively.

  19. Novel Campylobacter lari-like bacteria from humans and molluscs: description of Campylobacter peloridis sp. nov., Campylobacter lari subsp. concheus subsp. nov. and Campylobacter lari subsp. lari subsp. nov.

    Science.gov (United States)

    Debruyne, Lies; On, Stephen L W; De Brandt, Evie; Vandamme, Peter

    2009-05-01

    A polyphasic study was undertaken to clarify the taxonomic position of Campylobacter lari-like strains isolated from shellfish and humans. The diversity within the strain collection was initially screened by means of fluorescent amplified fragment length polymorphism analysis and whole-cell protein electrophoresis, revealing the existence of two clusters distinct from C. lari and other Campylobacter species. The divergence of these clusters was confirmed by phenotypic analysis and by 16S rRNA and hsp60 gene sequence analysis. Phylogenetic analysis identified C. lari, Campylobacter jejuni, Campylobacter coli and Campylobacter insulaenigrae as the closest phylogenetic neighbours of both taxa. DNA-DNA hybridizations revealed that one cluster, comprising 10 strains, represented a novel Campylobacter species, for which the name Campylobacter peloridis sp. nov. is proposed, with 2314BVA(T) (=LMG 23910(T) =CCUG 55787(T)) as the type strain. The second cluster, comprising six strains, represents a novel subspecies within the species C. lari, for which the name Campylobacter lari subsp. concheus subsp. nov. is proposed, with 2897R(T) (=LMG 21009(T) =CCUG 55786(T)) as the type strain. The description of C. lari subsp. concheus has the effect of automatically creating the subspecies Campylobacter lari subsp. lari subsp. nov. (type strain LMG 8846(T)=NCTC 11352(T)).

  20. Structural and functional studies of gpX of Escherichia coli phage P2 reveal a widespread role for LysM domains in the baseplates of contractile-tailed phages.

    Science.gov (United States)

    Maxwell, Karen L; Fatehi Hassanabad, Mostafa; Chang, Tom; Paul, Vivek D; Pirani, Nawaz; Bona, Diane; Edwards, Aled M; Davidson, Alan R

    2013-12-01

    A variety of bacterial pathogenicity determinants, including the type VI secretion system and the virulence cassettes from Photorhabdus and Serratia, share an evolutionary origin with contractile-tailed myophages. The well-characterized Escherichia coli phage P2 provides an excellent system for studies related to these systems, as its protein composition appears to represent the "minimal" myophage tail. In this study, we used nuclear magnetic resonance (NMR) spectroscopy to determine the solution structure of gpX, a 68-residue tail baseplate protein. Although the sequence and structure of gpX are similar to those of LysM domains, which are a large family associated with peptidoglycan binding, we did not detect a peptidoglycan-binding activity for gpX. However, bioinformatic analysis revealed that half of all myophages, including all that possess phage T4-like baseplates, encode a tail protein with a LysM-like domain, emphasizing a widespread role for this domain in baseplate function. While phage P2 gpX comprises only a single LysM domain, many myophages display LysM domain fusions with other tail proteins, such as the DNA circulation protein found in Mu-like phages and gp53 of T4-like phages. Electron microscopy of P2 phage particles with an incorporated gpX-maltose binding protein fusion revealed that gpX is located at the top of the baseplate, near the junction of the baseplate and tail tube. gpW, the orthologue of phage T4 gp25, was also found to localize to this region. A general colocalization of LysM-like domains and gpW homologues in diverse phages is supported by our bioinformatic analysis.

  1. Detection of Campylobacter jejuni and Campylobacter coli in Environmental Waters by PCR Enzyme-Linked Immunosorbent Assay

    OpenAIRE

    Sails, A. D.; Bolton, F. J.; Fox, A. J.; Wareing, D. R. A.; Greenway, D. L. A.

    2002-01-01

    A PCR enzyme-linked immunosorbent assay (ELISA) assay was applied to the detection of Campylobacter jejuni and Campylobacter coli in environmental water samples after enrichment culture. Bacterial cells were concentrated from 69 environmental water samples by using filtration, and the filtrates were cultured in Campylobacter blood-free broth. After enrichment culture, DNA was extracted from the samples by using a rapid-boiling method, and the DNA extracts were used as a template in a PCR ELIS...

  2. Twelve previously unknown phage genera are ubiquitous in global oceans.

    Science.gov (United States)

    Holmfeldt, Karin; Solonenko, Natalie; Shah, Manesh; Corrier, Kristen; Riemann, Lasse; Verberkmoes, Nathan C; Sullivan, Matthew B

    2013-07-30

    Viruses are fundamental to ecosystems ranging from oceans to humans, yet our ability to study them is bottlenecked by the lack of ecologically relevant isolates, resulting in "unknowns" dominating culture-independent surveys. Here we present genomes from 31 phages infecting multiple strains of the aquatic bacterium Cellulophaga baltica (Bacteroidetes) to provide data for an underrepresented and environmentally abundant bacterial lineage. Comparative genomics delineated 12 phage groups that (i) each represent a new genus, and (ii) represent one novel and four well-known viral families. This diversity contrasts the few well-studied marine phage systems, but parallels the diversity of phages infecting human-associated bacteria. Although all 12 Cellulophaga phages represent new genera, the podoviruses and icosahedral, nontailed ssDNA phages were exceptional, with genomes up to twice as large as those previously observed for each phage type. Structural novelty was also substantial, requiring experimental phage proteomics to identify 83% of the structural proteins. The presence of uncommon nucleotide metabolism genes in four genera likely underscores the importance of scavenging nutrient-rich molecules as previously seen for phages in marine environments. Metagenomic recruitment analyses suggest that these particular Cellulophaga phages are rare and may represent a first glimpse into the phage side of the rare biosphere. However, these analyses also revealed that these phage genera are widespread, occurring in 94% of 137 investigated metagenomes. Together, this diverse and novel collection of phages identifies a small but ubiquitous fraction of unknown marine viral diversity and provides numerous environmentally relevant phage-host systems for experimental hypothesis testing.

  3. Campylobacter jejuni acquire new host-derived CRISPR spacers when in association with bacteriophages harbouring a CRISPR-like Cas4 protein

    Directory of Open Access Journals (Sweden)

    Ian F. Connerton

    2015-01-01

    Full Text Available Campylobacter jejuni is a worldwide cause of human diarrhoeal disease. Clustered Repetitively Interspaced Palindromic Repeats (CRISPRs and associated proteins allow Bacteria and Archaea to evade bacteriophage and plasmid infection. Type II CRISPR systems are found in association with combinations of genes encoding the CRISPR-associated Cas1, Cas2, Cas4 or Csn2, and Cas9 proteins. C. jejuni possesses a minimal subtype II-C CRISPR system containing cas1, cas2, and cas9 genes whilst cas4 is notably absent. Cas4 proteins possess 5ʹ-3ʹ exonuclease activity to create recombinogenic-ends for spacer acquisition. Here we report a conserved Cas4-like protein in Campylobacter bacteriophages that creates a novel split arrangement between the bacteriophage and host that represents a new twist in the bacteriophage/host co-evolutionary arms race. The continuous association of bacteriophage and host in the carrier state life cycle of C. jejuni provided an opportunity to study spacer acquisition in this species. Remarkably all the spacer sequences observed were of host origin. We hypothesise that Campylobacter bacteriophages can use Cas4-like protein to activate spacer acquisition to use host DNA as an effective decoy to bacteriophage DNA. Bacteria that acquire self-spacers and escape phage infection must overcome CRISPR-mediated autoimmunity either by loss of the interference functions leaving them susceptible to foreign DNA incursion or tolerate changes in gene regulation.

  4. The Agricultural Antibiotic Carbadox Induces Phage-mediated Gene Transfer in Salmonella

    Directory of Open Access Journals (Sweden)

    Bradley L. Bearson

    2014-02-01

    Full Text Available Antibiotics are used for disease therapeutic or preventative effects in humans and animals, as well as for enhanced feed conversion efficiency in livestock. Antibiotics can also cause undesirable effects in microbial populations, including selection for antibiotic resistance, enhanced pathogen invasion, and stimulation of horizontal gene transfer. Carbadox is a veterinary antibiotic used in the U.S. during the starter phase of swine production for improved feed efficiency and control of swine dysentery and bacterial swine enteritis. Carbadox has been shown in vitro to induce phage-encoded Shiga toxin in Shiga toxin-producing Escherichia coli and a phage-like element transferring antibiotic resistance genes in Brachyspira hyodysenteriae, but the effect of carbadox on prophages in other bacteria is unknown. This study examined carbadox exposure on prophage induction and genetic transfer in Salmonella enterica serovar Typhimurium, a human foodborne pathogen that frequently colonizes swine without causing disease. S. Typhimurium LT2 exposed to carbadox induced prophage production, resulting in bacterial cell lysis and release of virions that were visible by electron microscopy. Carbadox induction of phage-mediated gene transfer was confirmed by monitoring the transduction of a sodCIII::neo cassette in the Fels-1 prophage from LT2 to a recipient Salmonella strain. Furthermore, carbadox frequently induced generalized transducing phages in multidrug-resistant phage type DT104 and DT120 isolates, resulting in the transfer of chromosomal and plasmid DNA that included antibiotic resistance genes. Our research indicates that exposure of Salmonella to carbadox induces prophages that can transfer virulence and antibiotic resistance genes to susceptible bacterial hosts. Carbadox-induced, phage-mediated gene transfer could serve as a contributing factor in bacterial evolution during animal production, with prophages being a reservoir for bacterial fitness

  5. The Prevalence of Antibiotic and Biocide Resistance Among Campylobacter coli and Campylobacter jejuni from Different Sources

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    Ana Mavri

    2012-01-01

    Full Text Available The increasing levels of antimicrobial resistance among foodborne bacteria are recognised as an important emerging public health problem. Reduced susceptibility to biocides also appears to be increasing. A potential concern is the possibility that the widespread use of biocides is responsible for the selection and maintenance of antibiotic-resistant bacteria. Here, we examine the prevalence of erythromycin, ciprofloxacin, triclosan, benzalkonium chloride, chlorhexidine diacetate, cetylpyridinium chloride, trisodium phosphate and sodium dodecyl sulphate resistance among 27 isolates of Campylobacter coli and 15 isolates of Campylobacter jejuni from food, animal, human and environmental water sources. These antimicrobial susceptibilities were determined by the broth microdilution method. In the 42 Campylobacter strains studied, different antibiotic resistance levels were seen. The resistance to erythromycin and ciprofloxacin was observed in 14.3 % of Campylobacter strains. A higher rate of erythromycin resistance and multi-resistance was observed among isolated C. coli than among C. jejuni strains. Similar situations were seen for triclosan. Conversely, the level of benzalkonium chloride resistance was higher in C. jejuni than in C. coli. No correlation between biocide and antibiotic resistance was observed. This study does not provide evidence to confirm that tolerance to biocides is connected to antibiotic resistance in Campylobacter.

  6. Novel plasmid conferring kanamycin and tetracycline resistance in the turkey-derived Campylobacter jejuni strain 11601MD.

    Science.gov (United States)

    Crespo, M D; Altermann, E; Olson, J; Miller, W G; Chandrashekhar, K; Kathariou, S

    2016-07-01

    In Campylobacter spp., resistance to the antimicrobials kanamycin and tetracycline is frequently associated with plasmid-borne genes. However, relatively few plasmids of Campylobacter jejuni have been fully characterized to date. A novel plasmid (p11601MD; 44,095nt) harboring tet(O) was identified in C. jejuni strain 11601MD, which was isolated from the jejunum of a turkey produced conventionally in North Carolina. Analysis of the p11601MD sequence revealed the presence of a high-GC content cassette with four genes that included tet(O) and a putative aminoglycoside transferase gene (aphA-3) highly similar to kanamycin resistance determinants. Several genes putatively involved in conjugative transfer were also identified on the plasmid. These findings will contribute to a better understanding of the distribution of potentially self-mobilizing plasmids harboring antibiotic resistance determinants in Campylobacter spp. from turkeys and other sources.

  7. Phages of lactic acid bacteria: the role of genetics in understanding phage-host interactions and their co-evolutionary processes.

    Science.gov (United States)

    Mahony, Jennifer; Ainsworth, Stuart; Stockdale, Stephen; van Sinderen, Douwe

    2012-12-20

    Dairy fermentations are among the oldest food processing applications, aimed at preservation and shelf-life extension through the use of lactic acid bacteria (LAB) starter cultures, in particular strains of Lactococcus lactis, Streptococcus thermophilus, Lactobacillus spp. and Leuconostoc spp. Traditionally this was performed by continuous passaging of undefined cultures from a finished fermentation to initiate the next fermentation. More recently, consumer demands on consistent and desired flavours and textures of dairy products have led to a more defined approach to such processes. Dairy (starter) companies have responded to the need to define the nature and complexity of the starter culture mixes, and dairy fermentations are now frequently based on defined starter cultures of low complexity, where each starter component imparts specific technological properties that are desirable to the product. Both mixed and defined starter culture approaches create the perfect environment for the proliferation of (bacterio)phages capable of infecting these LAB. The repeated use of the same starter cultures in a single plant, coupled to the drive towards higher and consistent production levels, increases the risk and negative impact of phage infection. In this review we will discuss recent advances in tracking the adaptation of phages to the dairy industry, the advances in understanding LAB phage-host interactions, including evolutionary and genomic aspects.

  8. A novel Pseudomonas aeruginosa bacteriophage, Ab31, a chimera formed from temperate phage PAJU2 and P. putida lytic phage AF: characteristics and mechanism of bacterial resistance.

    Directory of Open Access Journals (Sweden)

    Libera Latino

    Full Text Available A novel temperate bacteriophage of Pseudomonas aeruginosa, phage vB_PaeP_Tr60_Ab31 (alias Ab31 is described. Its genome is composed of structural genes related to those of lytic P. putida phage AF, and regulatory genes similar to those of temperate phage PAJU2. The virion structure resembles that of phage AF and other lytic Podoviridae (S. enterica Epsilon 15 and E. coli phiv10 with similar tail spikes. Ab31 was able to infect P. aeruginosa strain PA14 and two genetically related strains called Tr60 and Tr162, out of 35 diverse strains from cystic fibrosis patients. Analysis of resistant host variants revealed different phenotypes, including induction of pigment and alginate overproduction. Whole genome sequencing of resistant variants highlighted the existence of a large deletion of 234 kbp in two strains, encompassing a cluster of genes required for the production of CupA fimbriae. Stable lysogens formed by Ab31 in strain Tr60, permitted the identification of the insertion site. During colonization of the lung in cystic fibrosis patients, P. aeruginosa adapts by modifying its genome. We suggest that bacteriophages such as Ab31 may play an important role in this adaptation by selecting for bacterial characteristics that favor persistence of bacteria in the lung.

  9. Phages of lactic acid bacteria: The role of genetics in understanding phage-host interactions and their co-evolutionary processes

    Energy Technology Data Exchange (ETDEWEB)

    Mahony, Jennifer, E-mail: j.mahony@ucc.ie [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Ainsworth, Stuart; Stockdale, Stephen [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Sinderen, Douwe van, E-mail: d.vansinderen@ucc.ie [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Alimentary Pharmabiotic Centre, Biosciences Institute, University College Cork, Western Road, Cork (Ireland)

    2012-12-20

    Dairy fermentations are among the oldest food processing applications, aimed at preservation and shelf-life extension through the use of lactic acid bacteria (LAB) starter cultures, in particular strains of Lactococcus lactis, Streptococcus thermophilus, Lactobacillus spp. and Leuconostoc spp. Traditionally this was performed by continuous passaging of undefined cultures from a finished fermentation to initiate the next fermentation. More recently, consumer demands on consistent and desired flavours and textures of dairy products have led to a more defined approach to such processes. Dairy (starter) companies have responded to the need to define the nature and complexity of the starter culture mixes, and dairy fermentations are now frequently based on defined starter cultures of low complexity, where each starter component imparts specific technological properties that are desirable to the product. Both mixed and defined starter culture approaches create the perfect environment for the proliferation of (bacterio)phages capable of infecting these LAB. The repeated use of the same starter cultures in a single plant, coupled to the drive towards higher and consistent production levels, increases the risk and negative impact of phage infection. In this review we will discuss recent advances in tracking the adaptation of phages to the dairy industry, the advances in understanding LAB phage-host interactions, including evolutionary and genomic aspects.

  10. A PCR-RFLP assay for the detection and differentiation of Campylobacter jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis.

    Science.gov (United States)

    Kamei, Kazumasa; Asakura, Masahiro; Somroop, Srinuan; Hatanaka, Noritoshi; Hinenoya, Atsushi; Nagita, Akira; Misawa, Naoaki; Matsuda, Motoo; Nakagawa, Shinsaku; Yamasaki, Shinji

    2014-05-01

    Although Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of human gastrointestinal diseases, other Campylobacter species are also involved in human and animal infections. In this study, we developed a cytolethal distending toxin (cdt) gene-based PCR-RFLP assay for the detection and differentiation of C. jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis. Previously designed common primers, which can amplify the cdtB gene of C. jejuni, C. coli and C. fetus, were used for detecting seven Campylobacter species and differentiating between them by restriction digestion. The PCR-RFLP assay was validated with 277 strains, including 35 C. jejuni, 19 C. coli, 20 C. fetus, 24 C. hyointestinalis, 13 C. lari, 2 C. helveticus, 22 C. upsaliensis, 3 other Campylobacter spp. and 17 other species associated with human diseases. Sensitivity and specificity of the PCR-RFLP assay were 100 % except for C. hyointestinalis (88 % sensitivity). Furthermore, the PCR-RFLP assay successfully detected and differentiated C. jejuni, C. coli and C. fetus in clinical and animal samples. The results indicate that the PCR-RFLP assay is useful for the detection and differentiation of seven Campylobacter species important for human and animal diseases.

  11. Longitudinal study of the excretion patterns of thermophilic Campylobacter spp. in young pet dogs in Denmark

    DEFF Research Database (Denmark)

    Hald, Birthe; Pedersen, Karl; Wainø, Michael;

    2004-01-01

    .7% Campylobacter coli, and 2.8% Campylobacter spp. Isolates were typed by pulsed-field gel electrophoresis (PFGE) to elucidate the strain excretion pattern. All study dogs excreted Campylobacter spp. during the study period. At 3 months of age, 60% of the dogs carried Campylobacter, increasing to nearly 100...

  12. Automated Detection of Conformational Epitopes Using Phage Display Peptide Sequences

    Directory of Open Access Journals (Sweden)

    Surendra S Negi

    2009-01-01

    Full Text Available Background: Precise determination of conformational epitopes of neutralizing antibodies represents a key step in the rational design of novel vaccines. A powerful experimental method to gain insights on the physical chemical nature of conformational epitopes is the selection of linear peptides that bind with high affinities to a monoclonal antibody of interest by phage display technology. However, the structural characterization of conformational epitopes from these mimotopes is not straightforward, and in the past the interpretation of peptide sequences from phage display experiments focused on linear sequence analysis to find a consensus sequence or common sequence motifs.Results: We present a fully automated search method, EpiSearch that predicts the possible location of conformational epitopes on the surface of an antigen. The algorithm uses peptide sequences from phage display experiments as input, and ranks all surface exposed patches according to the frequency distribution of similar residues in the peptides and in the patch. We have tested the performance of the EpiSearch algorithm for six experimental data sets of phage display experiments, the human epidermal growth factor receptor-2 (HER-2/neu, the antibody mAb Bo2C11 targeting the C2 domain of FVIII, antibodies mAb 17b and mAb b12 of the HIV envelope protein gp120, mAb 13b5 targeting HIV-1 capsid protein and 80R of the SARS coronavirus spike protein. In all these examples the conformational epitopes as determined by the X-ray crystal structures of the antibody-antigen complexes, were found within the highest scoring patches of EpiSearch, covering in most cases more than 50% residues of experimental observed conformational epitopes. Input options of the program include mapping of a single peptide or a set of peptides on the antigen structure, and the results of the calculation can be visualized on our interactive web server.Availability: Users can access the EpiSearch from our web

  13. Dualities in the analysis of phage DNA packaging motors

    Science.gov (United States)

    Serwer, Philip; Jiang, Wen

    2012-01-01

    The DNA packaging motors of double-stranded DNA phages are models for analysis of all multi-molecular motors and for analysis of several fundamental aspects of biology, including early evolution, relationship of in vivo to in vitro biochemistry and targets for anti-virals. Work on phage DNA packaging motors both has produced and is producing dualities in the interpretation of data obtained by use of both traditional techniques and the more recently developed procedures of single-molecule analysis. The dualities include (1) reductive vs. accretive evolution, (2) rotation vs. stasis of sub-assemblies of the motor, (3) thermal ratcheting vs. power stroking in generating force, (4) complete motor vs. spark plug role for the packaging ATPase, (5) use of previously isolated vs. new intermediates for analysis of the intermediate states of the motor and (6) a motor with one cycle vs. a motor with two cycles. We provide background for these dualities, some of which are under-emphasized in the literature. We suggest directions for future research. PMID:23532204

  14. Structure, adsorption to host, and infection mechanism of virulent lactococcal phage p2.

    Science.gov (United States)

    Bebeacua, Cecilia; Tremblay, Denise; Farenc, Carine; Chapot-Chartier, Marie-Pierre; Sadovskaya, Irina; van Heel, Marin; Veesler, David; Moineau, Sylvain; Cambillau, Christian

    2013-11-01

    Lactococcal siphophages from the 936 and P335 groups infect the Gram-positive bacterium Lactococcus lactis using receptor binding proteins (RBPs) attached to their baseplate, a large multiprotein complex at the distal part of the tail. We have previously reported the crystal and electron microscopy (EM) structures of the baseplates of phages p2 (936 group) and TP901-1 (P335 group) as well as the full EM structure of the TP901-1 virion. Here, we report the complete EM structure of siphophage p2, including its capsid, connector complex, tail, and baseplate. Furthermore, we show that the p2 tail is characterized by the presence of protruding decorations, which are related to adhesins and are likely contributed by the major tail protein C-terminal domains. This feature is reminiscent of the tail of Escherichia coli phage λ and Bacillus subtilis phage SPP1 and might point to a common mechanism for establishing initial interactions with their bacterial hosts. Comparative analyses showed that the architecture of the phage p2 baseplate differs largely from that of lactococcal phage TP901-1. We quantified the interaction of its RBP with the saccharidic receptor and determined that specificity is due to lower k(off) values of the RBP/saccharidic dissociation. Taken together, these results suggest that the infection of L. lactis strains by phage p2 is a multistep process that involves reversible attachment, followed by baseplate activation, specific attachment of the RBPs to the saccharidic receptor, and DNA ejection.

  15. Assembling filamentous phage occlude pIV channels.

    Science.gov (United States)

    Marciano, D K; Russel, M; Simon, S M

    2001-07-31

    Filamentous phage f1 is exported from its Escherichia coli host without killing the bacterial cell. Phage-encoded protein pIV, which is required for phage assembly and secretion, forms large highly conductive channels in the outer membrane of E. coli. It has been proposed that the phage are extruded across the bacterial outer membrane through pIV channels. To test this prediction, we developed an in vivo assay by using a mutant pIV that functions in phage export but whose channel opens in the absence of phage extrusion. In E. coli lacking its native maltooligosacharride transporter LamB, this pIV variant allowed oligosaccharide transport across the outer membrane. This entry of oligosaccharide was decreased by phage production and still further decreased by production of phage that cannot be released from the cell surface. Thus, exiting phage block the pIV-dependent entry of oligosaccharide, suggesting that phage occupy the lumen of pIV channels. This study provides the first evidence, to our knowledge, for viral exit through a large aqueous channel.

  16. A Lower-Order Oligomer Form of Phage Shock Protein A (PspA) Stably Associates with the Hexameric AAA+ Transcription Activator Protein PspF for Negative Regulation

    OpenAIRE

    Joly, Nicolas; Burrows, Patricia C.; Engl, Christoph; Jovanovic, Goran; Buck, Martin

    2009-01-01

    To survive and colonise their various environments, including those used during infection, bacteria have developed a variety of adaptive systems. Amongst these is phage shock protein (Psp) response, which can be induced in Escherichia coli upon filamentous phage infection (specifically phage secretin pIV) and by other membrane-damaging agents. The E. coli Psp system comprises seven proteins, of which PspA is the central component. PspA is a bifunctional protein that is directly involved in (i...

  17. Antimicrobial resistance in Campylobacter coli and Campylobacter jejuni in cynomolgus monkeys (Macaca fascicularis) and eradication regimens.

    Science.gov (United States)

    Koga, Tetsufumi; Aoki, Wataru; Mizuno, Takashi; Wakazono, Kuniko; Ohno, Junki; Nakai, Tsunehiro; Nomiya, Takao; Fujii, Miki; Fusegawa, Keiichi; Kinoshita, Kazuya; Hamada, Takakazu; Ikeda, Yoshinori

    2017-02-01

    Campylobacter spp. are zoonotic pathogens, however, knowledge about their presence and antimicrobial resistance in nonhuman primates is limited. Our animal facility purchased cynomolgus monkeys (Macaca fascicularis) from various Asian countries: China, Cambodia, Indonesia, the Philippines, and Vietnam. Colonization by Campylobacter spp. was investigated in 238 of the monkeys from 2009 to 2012 and antimicrobial susceptibility testing was carried out for these isolates. Furthermore, we eradicated these pathogens from these monkeys. Campylobacter spp. were isolated from 47 monkeys from three specific countries: China, Cambodia, and Indonesia, with respective isolation rates of 15%, 36%, and 67%. Two monkeys, which were each infected with Campylobacter jejuni and Campylobacter coli, showed clinical symptoms of diarrhea and bloody feces. In total, 41 isolates of C. coli and 17 isolates of C. jejuni were detected. Antimicrobial susceptibility varied: in the monkeys from China, erythromycin (ERY)-, tetracycline (TET)-, and ciprofloxacin-resistant C. coli, in the monkeys from Cambodia, amoxicillin-intermediate, TET- and ciprofloxacin-resistant C. coli and amoxicillin- and ciprofloxacin-resistant C. jejuni, and in the monkeys from Indonesia, ciprofloxacin-resistant C. coli and TET- and ciprofloxacin-resistant C. jejuni were common (>75%). Multiresistant isolates of C. coli were found in monkeys from all countries and multiresistant isolates of C. jejuni were found in monkeys from Indonesia. The eradication rate with azithromycin was comparable to that with gentamicin (GEN) by oral administration, and was higher than those with amoxicillin-clavulanic acid (AMC) and chloramphenicol (CHL). From the perspective of zoonosis, we should acknowledge multiresistant Campylobacter spp. isolated from the monkeys as a serious warning. Copyright © 2015. Published by Elsevier B.V.

  18. Combined Campylobacter jejuni and Campylobacter coli Rapid Testing and Molecular Epidemiology in Conventional Broiler Flocks.

    Science.gov (United States)

    Schallegger, G; Muri-Klinger, S; Brugger, K; Lindhardt, C; John, L; Glatzl, M; Wagner, M; Stessl, B

    2016-12-01

    Campylobacter spp. are important causes of bacterial zoonosis, most often transmitted by contaminated poultry meat. From an epidemiological and risk assessment perspective, further knowledge should be obtained on Campylobacter prevalence and genotype distribution in primary production. Consequently, 15 Austrian broiler flocks were surveyed in summer for their thermophilic Campylobacter spp. contamination status. Chicken droppings, dust and drinking water samples were collected from each flock at three separate sampling periods. Isolates were confirmed by PCR and subtyped. We also compared three alternative methods (culture-based enrichment in Bolton broth, culture-independent real-time PCR and a lateral-flow test) for their applicability in chicken droppings. Twelve flocks were found to be positive for thermophilic Campylobacter spp. during the entire sampling period. Seven flocks (46.6%) were contaminated with both, C. jejuni and C. coli, five flocks harboured solely one species. We observed to a majority flock-specific C. jejuni and C. coli genotypes, which dominated the respective flock. Flocks within a distance jejuni genotypes indicating a cross-contamination event via the environment or personnel vectors. Multilocus sequence typing (MLST) of C. jejuni revealed that the majority of isolates were assigned to globally distributed clonal complexes or had a strong link to the human interface (CC ST-446 and ST4373). The combination of techniques poses an advantage over risk assessment studies based on cultures alone, as, in the case of Campylobacter, occurrence of a high variety of genotypes might be present among a broiler flock. We suggest applying the lateral-flow test under field conditions to identify 'high-shedding' broiler flocks at the farm level. Consequently, poultry farmers and veterinarians could improve hygiene measurements and direct sanitation activities, especially during the thinning period. Ultimately, real-time PCR could be applied to quantify

  19. Morphology, genome sequence, and structural proteome of type phage P335 from Lactococcus lactis

    DEFF Research Database (Denmark)

    Labrie, Simon J.; Josephsen, Jytte; Neve, Horst;

    2008-01-01

    Lactococcus lactis phage P335 is a virulent type phage for the species that bears its name and belongs phage P335 is a virulent type phage for the species that bears its name and belongs to the Siphoviridae family. Morphologically, P335 resembled the L. lactis phages TP901-1 and Tuc2009, except...

  20. Campylobacter fetus bacteremia in an immunocompetent traveler.

    Science.gov (United States)

    Mikals, Kyle; Masel, Jennifer; Gleeson, Todd

    2014-10-01

    Campylobacter fetus bacteremia is a rare human infection that occurs almost exclusively in the setting of advanced age, immunosuppression, human immunodeficiency virus infection, alcoholism, or recent gastrointestinal surgery. This report of C. fetus bacteremia in a 39-year-old immunocompetent traveler who ate raw beef identifies C. fetus as a potential emerging pathogen in normal hosts. © The American Society of Tropical Medicine and Hygiene.

  1. Prevalence and Antimicrobial Resistance of Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Salehi

    2014-07-01

    Full Text Available Background Campylobacter spp. are Gram-negative bacilli enteric pathogens that pose a major public health problem worldwide. In this genus, the most important species is Campylobacter jejuni. This bacterium causes diarrhea as its main symptom, which its intensity varies from mild to severe. Patients’ stools may be watery or bloody. Objectives In this study, we aimed to determine the prevalence of the species of Campylobacter. jejuni in Zahedan, a major city in southeastern Iran. Patients and Methods Fecal samples from 164 patients with acute diarrhea from Zahedan hospitals were collected from 2011 to 2013. Then the samples were streaked onto a campylobacter selective agar containing supplement and 7% defibrinated sheep blood. Conventional bacteriological tests (such as culture and biochemical tests were performed to confirm the genus and differentiate at the species level. Finally, disk diffusion method was performed according to the recommendation of Clinical and Laboratory Standards Institute (CLSI to determine the susceptibility of isolates to antibacterial agents. Results Out of 164 samples, 19 (11.6% were reported positive by culture which confirmed by biochemical tests. Fifteen (78.9% patients, whose samples were positive, hospitalized in infant ward. Two (10.5% patients treated as outpatients. Two remaining (10.5% patients were admitted in internal medicine ward. All of isolated strains were susceptible or moderately susceptible to erythromycin as the drug of choice. Conclusions In this study, the prevalence of the disease (11.6% is found to be more than other parts of Iran. The symptomatic infection mainly affects children younger than 5 years.

  2. Isolation of Campylobacter from human stool samples

    Directory of Open Access Journals (Sweden)

    S M Salim

    2014-01-01

    Full Text Available Context: Campylobacter is an undetected cause of diarrhoea especially under 5 years of age in most of the countries. Isolation of this organism is difficult, expensive and cumbersome. Aims: Our objective of this study was to isolate this pathogen from the stool specimens on routinely available blood containing laboratory media using the candle jar for creating the microaerophilic atmosphere in our setup. Settings and Designs: A descriptive study. Materials and Methods: A total of 50 stool samples were inoculated onto selective and non-selective media with and without filtration using a 0.45 μm membrane. The inoculated media were simultaneously incubated in microaerophilic conditions using the Anoxomat as well as in candle jars at temperatures 37°C and 42°C. The culture isolates were confirmed by standard phenotypic tests. A simplex polymerase chain reaction (PCR targeting the 16S ribosomal deoxyribonucleic acid of Campylobacter was performed on the deoxyribonucleic acid (DNA of the culture isolates as well as on the DNA extracted from the stool filtrates. Statistical Analysis: Data was expressed as a proportion. Results: Campylobacter could be isolated in 5 out of 50 stool samples using both the Anoxomat as well as the candle jar. Furthermore, we did not find any difference between the isolation using the selective and blood containing media as well as the different incubation temperatures. All the five were confirmed phenotypically and genotypically to be Campylobacter jejuni. The PCR results corroborated with that of the culture. Conclusions: Isolation by culture was as sensitive as that of the PCR.

  3. Glucose Metabolism via the Entner-Doudoroff Pathway in Campylobacter

    DEFF Research Database (Denmark)

    Vegge, Christina S; Jansen van Rensburg, Melissa J; Rasmussen, Janus J;

    2016-01-01

    Isolates of the zoonotic pathogen Campylobacter are generally considered to be unable to metabolize glucose due to lack of key glycolytic enzymes. However, the Entner-Doudoroff (ED) pathway has been identified in Campylobacter jejuni subsp. doylei and a few C. coli isolates. A systematic search f...

  4. Campylobacter infections in fattening pigs; excretion pattern and genetic diversity

    NARCIS (Netherlands)

    Weijtens, M.J.B.M.; Reinders, R.D.; Urlings, H.A.P.; Plas, van der J.

    1999-01-01

    The excretion of campylobacter by eight individually housed fattening pigs was monitored during 15 weeks. Rectal faeces samples were collected six times from these pigs and twice from their mothers (seven sows). Campylobacter was cultured from these samples on Preston medium. In some pigs, samples p

  5. Foodborne disease prevention and broiler chickens with reduced Campylobacter infection

    DEFF Research Database (Denmark)

    Bahrndorff, Simon; Rangstrup-Christensen, Lena; Nordentoft, Steen

    2013-01-01

    Studies have suggested that flies play a linking role in the epidemiology of Campylobacter spp. in broiler chickens and that fly screens can reduce the prevalence of Campylobacter spp. We examined the year-round and long-term effects of fly screens in 10 broiler chicken houses (99 flocks) in Denm...

  6. The role of Campylobacter jejuni cytolethal distending toxin in gastroenteritis

    DEFF Research Database (Denmark)

    Mortensen, Ninell P; Schiellerup, Peter; Boisen, Nadia

    2011-01-01

    The role of Campylobacter jejuni cytolethal distending toxin (CDT) on clinical outcome after gastroenteritis was investigated. Clinical data, blood serum samples, and Campylobacter spp. isolated, from each of 30 patients were collected over a period of 6 months. The CDT encoding genes, cdt...

  7. Cellular response of Campylobacter jejuni to trisodium phosphate

    DEFF Research Database (Denmark)

    Riedel, Charlotte Tandrup; Cohn, M. T.; Stabler, R. A.

    2012-01-01

    The highly alkaline compound trisodium phosphate (TSP) is used as an intervention to reduce the load of Campylobacter on poultry meat in U.S. poultry slaughter plants. The aim of the present study was to investigate the cellular responses of Campylobacter jejuni NCTC11168 when exposed to sublethal...

  8. Campylobacter infections in fattening pigs; Excretion pattern and genetic diversity

    NARCIS (Netherlands)

    Weijtens, M.J.B.M.; Reinders, R.D.; Urlings, H.A.P.; Plas, J. van der

    1999-01-01

    The excretion of campylobacter by eight individually housed fattening pigs was monitored during 15 weeks. Rectal faeces samples were collected six times from these pigs and twice from their mothers (seven sows). Campylobacter was cultured from these samples on Preston medium. In some pigs, samples

  9. Campylobacter and Toll-like receptors : implications for vaccine development

    NARCIS (Netherlands)

    de Zoete, M.R.|info:eu-repo/dai/nl/30483419X

    2010-01-01

    Campylobacter jejuni is a Gram-negative highly motile bacterium that colonizes the intestinal tract of humans, leading to inflammation of the intestinal mucosal layer. Campylobacter-induced enteritis causes (bloody) diarrhea, cramps, malaise and fever, which resolves within two weeks. In a small

  10. Effectiveness and Efficiency of Controlling Campylobacter on Broiler Chicken Meat

    NARCIS (Netherlands)

    Havelaar, A.H.; Mangen, M.J.J.; Koeijer, de A.A.; Bogaardt, M.J.; Evers, E.G.; Jacobs-Reitsma, W.F.; Pelt, van W.; Wagenaar, J.A.; Wit, de G.A.; Zee, van der H.; Nauta, M.J.

    2007-01-01

    Campylobacter bacteria are an important cause of foodborne infections. We estimated the potential costs and benefits of a large number of possible interventions to decrease human exposure to Campylobacter by consumption of chicken meat, which accounts for 20¿40% of all cases of human campylobacterio

  11. Preventing Campylobacter at the Source: Why Is It So Difficult?

    NARCIS (Netherlands)

    Wagenaar, J.A.; French, N.P.; Havelaar, A.H.

    2013-01-01

    Campylobacteriosis in humans, caused by Campylobacter jejuni and Campylobacter coli, is the most common recognized bacterial zoonosis in the European Union and the United States. The acute phase is characterized by gastrointestinal symptoms. The long-term sequelae (Guillain-Barre syndrome, reactive

  12. Quantifying Transmission of Campylobacter jejuni in Commercial Broiler Flocks

    NARCIS (Netherlands)

    Gerwe, van T.; Miflin, J.K.; Templeton, J.M.; Bouma, A.; Wagenaar, J.A.; Jacobs-Reitsma, W.F.; Stegeman, A.; Klinkenberg, D.

    2009-01-01

    Since meat from poultry colonized with Campylobacter spp. is a major cause of bacterial gastroenteritis, human exposure should be reduced by, among other things, prevention of colonization of broiler flocks. To obtain more insight into possible sources of introduction of Campylobacter into broiler f

  13. A Review of Bacteriocins to Control Campylobacter spp. in Poultry

    Science.gov (United States)

    The unacceptably high frequency of Campylobacter jejuni transmission from poultry to humans encourages scientists to consider and create alternative intervention strategies to control the pathogen in poultry production. Extremely high numbers of Campylobacter (often >108 cfu/g of poultry intestinal...

  14. Bacteriocins to Control Campylobacter spp. in Poultry—a Review

    Science.gov (United States)

    The unacceptably high frequency of Campylobacter jejuni transmission from poultry to humans encourages scientists to consider and create alternative intervention strategies to control the pathogen in poultry production. Extremely high numbers of Campylobacter (often >108 cfu/g of poultry intestinal...

  15. Campylobacter infections in fattening pigs; Excretion pattern and genetic diversity

    NARCIS (Netherlands)

    Weijtens, M.J.B.M.; Reinders, R.D.; Urlings, H.A.P.; Plas, J. van der

    1999-01-01

    The excretion of campylobacter by eight individually housed fattening pigs was monitored during 15 weeks. Rectal faeces samples were collected six times from these pigs and twice from their mothers (seven sows). Campylobacter was cultured from these samples on Preston medium. In some pigs, samples p

  16. Efficacy of natural cranberry extracts against campylobacter colonization in poultry

    Science.gov (United States)

    Campylobacter spp. has been identified as one of the leading causative agents of food borne diarrheal illness. Epi-demiological evidence has shown that poultry is the main source for human infection. Currently there are no consistently effective treatments to eliminate Campylobacter from poultry flo...

  17. Antimicrobial resistance of Campylobacter: prevalence and trends in Japan.

    Science.gov (United States)

    Igimi, S; Okada, Y; Ishiwa, A; Yamasaki, M; Morisaki, N; Kubo, Y; Asakura, H; Yamamoto, S

    2008-09-01

    Campylobacter is one of the most frequently diagnosed bacterial causes of human gastroenteritis in Japan and throughout the world. Resistance to quinolones in Campylobacter jejuni and C. coli isolated from humans has emerged in many countries during the past 15 years because fluoroquinolones are the drug of choice for the treatment of suspected bacterial gastroenteritis. Food contaminated with Campylobacter is the usual source of human infection; therefore, the presence of antimicrobial resistance strains in the food chain has raised concerns that the treatment of human infections will be compromised. The use of antimicrobial agents for food animals and in veterinary medicine is suspected to be correlated with an increase in quinolone-resistant strains of Campylobacter in food animals, especially in poultry products. In contrast to macrolide resistance in C. jejuni and C. coli isolated from humans showing a stable low rate, resistant Campylobacter spp. to quinolones have emerged in Japan. The paper summarizes food-borne Campylobacter infection in Japan, and the prevalence and trends of antimicrobial resistance of Campylobacter from the authors' data and other Japanese papers which reported the antimicrobial resistance of Campylobacter.

  18. Campylobacter and Toll-like receptors : implications for vaccine development

    NARCIS (Netherlands)

    de Zoete, M.R.

    2010-01-01

    Campylobacter jejuni is a Gram-negative highly motile bacterium that colonizes the intestinal tract of humans, leading to inflammation of the intestinal mucosal layer. Campylobacter-induced enteritis causes (bloody) diarrhea, cramps, malaise and fever, which resolves within two weeks. In a small num

  19. Campylobacter and Toll-like receptors : implications for vaccine development

    NARCIS (Netherlands)

    de Zoete, M.R.

    2010-01-01

    Campylobacter jejuni is a Gram-negative highly motile bacterium that colonizes the intestinal tract of humans, leading to inflammation of the intestinal mucosal layer. Campylobacter-induced enteritis causes (bloody) diarrhea, cramps, malaise and fever, which resolves within two weeks. In a small num

  20. Prevalence, Antimicrobial Resistance and Risk Factors for Thermophilic Campylobacter Infections in Symptomatic and Asymptomatic Humans in Tanzania.

    Science.gov (United States)

    Komba, E V G; Mdegela, R H; Msoffe, P L M; Nielsen, L N; Ingmer, H

    2015-11-01

    The genus Campylobacter comprises members known to be a leading cause of foodborne gastrointestinal illness worldwide. A study was conducted to determine the epidemiology and antimicrobial resistance of Campylobacter in humans in Morogoro, Eastern Tanzania. Isolation of Campylobacter from stool specimens adopted the Cape Town protocol. Campylobacter isolates were preliminarily identified by conventional phenotypic tests and subsequently confirmed by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry and polymerase chain reaction. Antimicrobial resistance testing employed the disc diffusion method. A small proportion of the test isolates was also subjected to agar dilution method. Risk factors for human illness were determined in an unmatched case-control study. Thermophilic Campylobacter were isolated from 11.4% of the screened individuals (n = 1195). The agreement between PCR and MALDI-TOF was perfect (κ = 1.0). Symptomatics and young individuals were infected with higher numbers than asymptomatic and adults, respectively. The majority (84.6%) of the isolates were C. jejuni and the remaining were C. coli. Isolates had highest resistance (95.6%) for colistin sulphate and lowest for ciprofloxacin (22.1%). The rates of resistance for other antibiotics (azithromycin, erythromycin, tetracycline, cephalothin, gentamycin, nalidixic acid, ampicillin, amoxycillin, norfloxacin, chloramphenicol) ranged from 44.1% to 89%. Comparison between disc diffusion and agar dilution methods indicated a good correlation, and the tests were in agreement to each other (κ ≥ 0.75). Human illness was found to be associated with young age and consumption of chicken meat and pre-prepared salad. Our data indicate the presence of antibiotic-resistant thermophilic Campylobacter in humans in the study area. There is a need for routine investigation of the presence of the organisms in gastroenteritis aetiology, including determination of their antibiotic

  1. Phage therapy of staphylococcal chronic osteomyelitis in experimental animal model

    Directory of Open Access Journals (Sweden)

    Chandan Kishor

    2016-01-01

    Full Text Available Background & objectives: Methicillin resistant Staphylococcus aureus (MRSA are the commonest cause of osteomyelitis. The aim of this study was to evaluate the role of an alternative therapy i.e. application of S. aureus specific bacteriophages in cases of osteomyelitis caused by MRSA in animal model. Methods: Twenty two rabbits were included in this study. The first two rabbits were used to test the safety of phage cocktail while the remaining 20 rabbits were divided into three groups; group A (n=4 to assess the establishment of osteomyelitis; group B (n=4 osteomyelitis developed but therapy started only after six weeks; and group C (n=12 osteomyelitis developed and therapy started after three weeks. Groups B and C rabbits were treated with four doses of cocktail of seven virulent bacteriophages at the interval of 48 h. Comparison between three groups was made on the basis of observation of clinical, radiological, microbiological, and histopathological examinations. Results: Experimental group rabbits recovered from the illness in the subsequent two weeks of the therapy. Appetite and activity of the rabbits improved, local oedema, erythema and induration subsided. There were minimal changes associated with osteomyelitis in X-ray and histopathology also showed no signs of infection with new bone formation. Control B group rabbits also recovered well from the infection. Interpretation & conclusions: The present study shows a potential of phage therapy to treat difficult infections caused by multidrug resistant bacteria.

  2. Sites and gene products involved in lambdoid phage DNA packaging.

    Science.gov (United States)

    Smith, M P; Feiss, M

    1993-04-01

    21 is a temperate lambdoid coliphage, and the genes that encode the head proteins of lambda and 21 are descended from a common ancestral bacteriophage. The sequencing of terminase genes 1 and 2 of 21 was completed, along with that of a segment at the right end of 21 DNA that includes the R4 sequence. The R4 sequence, a site that is likely involved in termination of DNA packaging, was found to be very similar to the R4 sequences of lambda and phi 80, suggesting that R4 is a recognition site that is not phage specific. DNA packaging by 21 is dependent on a host protein, integration host factor. A series of mutations in gene 1 (her mutations), which allow integration host factor-independent DNA packaging by 21, were found to be missense changes that affect predicted alpha-helixes in gp1. gp2, the large terminase subunit, is predicted to contain an ATP-binding domain and, perhaps, a second domain important for the cos-cutting activity of terminase. orf1, an open reading frame analogous in position to FI, a lambda gene involved in DNA packaging, shares some sequence identity with FI. orf1 was inactivated with nonsense and insertion mutations; these mutations were found not to affect phage growth. 21 was also not able to complement a lambda FI mutant.

  3. Phage therapy: delivering on the promise.

    Science.gov (United States)

    Harper, D R; Anderson, J; Enright, M C

    2011-07-01

    Bacteriophages are viruses that infect and, in many cases, destroy their bacterial targets. Within a few years of their initial discovery they were being investigated as therapeutic agents for infectious disease, an approach known as phage therapy. However, the nature of these exquisitely specific agents was not understood and much early use was both uninformed and unsuccessful. As a result they were replaced by chemical antibiotics once these became available. Although work on phage therapy continued (and continues) in Eastern Europe, this was not conducted to a standard allowing it to support clinical uses in areas regulated by the European Medicines Agency or the US FDA. To develop phage therapy for these areas requires work carried out in accordance with the requirements of these agencies, and, driven by the current crisis of antibiotic resistance, such clinical trials are now under way. The first Phase I clinical trial of safety was reported in 2005, and the results of the first Phase II clinical trial of efficacy of a bacteriophage therapeutic was published in 2009. While the delivery of these relatively large and complex agents to the site of disease can be more challenging than for conventional, small-molecule antibiotics, bacteriophages are then able to multiply locally even from an extremely low (picogram range) initial dose. This multiplication where and only where they are needed underlies the potential for bacteriophage therapeutics to become a much needed and powerful weapon against bacterial disease.

  4. A comparison of risk assessments on Campylobacter in broiler meat

    DEFF Research Database (Denmark)

    Nauta, Maarten; Hill, Andy; Rosenquist, Hanne

    2009-01-01

    In recent years. several quantitative risk assessments for Campylobacter in broiler meat have been developed to support risk managers in controlling this pathogen. The models encompass some or all of the consecutive stages in the broiler meat production chain: primary production, industrial...... of Campylobacter in flocks implies that monitoring for Campylobacter at the farm up to one week before slaughter may result in flocks that are falsely tested negative: once Campylobacter is established at the farm, the within-flock prevalence increases dramatically within a week. Consequently, at the point...... at reducing the Campylobacter concentration, rather than reducing the prevalence. During the stage where the consumer handles the food, cross-contamination is generally considered to be more relevant than undercooking. An important finding, shared by all, is that the tails of the distributions describing...

  5. Quantification of thermophilic Campylobacter spp. in broilers during meat processing.

    Science.gov (United States)

    Klein, Günter; Reich, Felix; Beckmann, Lutz; Atanassova, Viktoria

    2007-10-01

    Campylobacter spp. is a common cause of gastrointestinal illness. Since animal products, especially poultry meat, are an important source of human outbreaks of campylobacteriosis, tracing back to processing and initial production is of great interest. Samples were collected at a German poultry slaughterhouse for the estimation of the prevalence of Campylobacter at different processing steps. Quantification of Campylobacter in each of the samples was also performed. Out of 99 samples examined, 51 (51.5%) were positive for Campylobacter, with bacterial counts ranging from log(10) 6.5 cfu sample(-1) for carcasses to log 3.6 cfu ml(-1) for scalding water. The Campylobacter isolates (n = 51) were subtyped by pulsed-field gel electrophoresis using SmaI and KpnI restriction enzymes. Molecular typing showed a multitude of strains with different molecular patterns. Strains found in cloacal swabs before processing could also be isolated from carcasses at different processing steps.

  6. Exploring the risks of phage application in the environment

    Directory of Open Access Journals (Sweden)

    Sean eMeaden

    2013-11-01

    Full Text Available Interest in using bacteriophages to control the growth and spread of bacterial pathogens is being revived in the wake of widespread antibiotic resistance. However, little is known about the ecological effects that high concentrations of phages in the environment might have on natural microbial communities. We review the current evidence suggesting phage-mediated environmental perturbation, with a focus on agricultural examples, and describe the potential implications for human health and agriculture. Specifically, we examine the known and potential consequences of phage application in certain agricultural practices, discuss the risks of evolved bacterial resistance to phages, and question whether the future of phage therapy will emulate that of antibiotic treatment in terms of widespread resistance. Finally, we propose some basic precautions that could preclude such phenomena and highlight existing methods for tracking bacterial resistance to phage therapeutic agents.

  7. Pitfalls to avoid when using phage display for snake toxins.

    Science.gov (United States)

    Laustsen, Andreas Hougaard; Lauridsen, Line Præst; Lomonte, Bruno; Andersen, Mikael Rørdam; Lohse, Brian

    2017-02-01

    Antivenoms against bites and stings from snakes, spiders, and scorpions are associated with immunological side effects and high cost of production, since these therapies are still derived from the serum of hyper-immunized production animals. Biotechnological innovations within envenoming therapies are thus warranted, and phage display technology may be a promising avenue for bringing antivenoms into the modern era of biologics. Although phage display technology represents a robust and high-throughput approach for the discovery of antibody-based antitoxins, several pitfalls may present themselves when animal toxins are used as targets for phage display selection. Here, we report selected critical challenges from our own phage display experiments associated with biotinylation of antigens, clone picking, and the presence of amber codons within antibody fragment structures in some phage display libraries. These challenges may be detrimental to the outcome of phage display experiments, and we aim to help other researchers avoiding these pitfalls by presenting their solutions.

  8. Amplified fragment length polymorphism based identification of genetic markers and novel PCR assay for differentiation of Campylobacter fetus subspecies

    NARCIS (Netherlands)

    Bergen, van M.A.P.; Simons, G.; Graaf-van Bloois, van der L.; Putten, van J.P.; Rombout, J.; Wesley, I.; Wagenaar, J.A.

    2005-01-01

    Differentiation of Campylobacter fetus into C. fetus subsp. fetus (Cff) and C. fetus subsp. venerealis (Cfv) is important for both clinical and economic reasons. In the past, several molecular typing methods have been used for differentiation, including amplified fragment length polymorphism (AFLP).

  9. Lysogenic Conversion and Phage Resistance Development in Phage Exposed Escherichia coli Biofilms

    Directory of Open Access Journals (Sweden)

    Abram Aertsen

    2013-01-01

    Full Text Available In this study, three-day old mature biofilms of Escherichia coli were exposed once to either a temperate Shiga-toxin encoding phage (H-19B or an obligatory lytic phage (T7, after which further dynamics in the biofilm were monitored. As such, it was found that a single dose of H-19B could rapidly lead to a near complete lysogenization of the biofilm, with a subsequent continuous release of infectious H-19B particles. On the other hand, a single dose of T7 rapidly led to resistance development in the biofilm population. Together, our data indicates a profound impact of phages on the dynamics within structured bacterial populations.

  10. Relationship between Presence of Anti-Campylobacter FliD Protein Antibodies and Campylobacter jejuni Isolation from Broiler Chickens

    Science.gov (United States)

    Campylobacter jejuni, a Gram-negative rod, is a zoonotic pathogen associated with human acute bacterial gastroenteritis worldwide. Poultry products are regarded as a major source of this bacterium for human infection. Although this bacterium is a commensal in chicken cecal microbiome, Campylobacte...

  11. Risk factors for indigenous Campylobacter jejuni and Campylobacter coli infections in The Netherlands: a case-control study

    NARCIS (Netherlands)

    Doorduyn, Y.; Brandhof, van den W.E.; Duynhoven, van Y.T.H.P.; Breukink, B.J.; Wagenaar, J.A.; Pelt, van W.

    2010-01-01

    A case-control study comprising 1315 Campylobacter jejuni cases, 121 Campylobacter coli cases and 3409 frequency-matched controls was conducted in The Netherlands in 2002-2003. Risk factors for both C. jejuni and C. coli enteritis were consumption of undercooked meat and barbecued meat, ownership of

  12. Detection of Campylobacter Bacteria in Air Samples for Continuous Real-Time Monitoring of Campylobacter Colonization in Broiler Flocks

    DEFF Research Database (Denmark)

    Olsen, Katja Nyholm; Lund, Marianne; Skov, J.

    2009-01-01

    Improved monitoring tools are important for the control of Campylobacter bacteria in broiler production. In this study, we compare the sensitivities of detection of Campylobacter by PCR with feces, dust, and air samples during the lifetimes of broilers in two poultry houses and conclude that the ...

  13. Probing Tumor Microenvironment With In Vivo Phage Display

    Science.gov (United States)

    2014-10-01

    technology. A, Peptides found in the “Phage pool alone” group are listed in a descending order of frequency. Note that CISQERGESC (CIS) and CIFSGEGESC ( CIF ...expressing CISQERGESC (CIS: 0.6% of the recovered phage clones) and 202 phages expressing a relevant peptide CIFSGEGESC ( CIF : 0.2%) (Table 1A). The...term ended showed that CIS and CIF , which have very similar amino acid sequences, bind to cultured hb6011 CAFs especially to filopodia and fibrous

  14. Evolutionary Rationale for Phages as Complements of Antibiotics.

    Science.gov (United States)

    Torres-Barceló, Clara; Hochberg, Michael E

    2016-04-01

    Antibiotic-resistant bacterial infections are a major concern to public health. Phage therapy has been proposed as a promising alternative to antibiotics, but an increasing number of studies suggest that both of these antimicrobial agents in combination are more effective in controlling pathogenic bacteria than either alone. We advocate the use of phages in combination with antibiotics and present the evolutionary basis for our claim. In addition, we identify compelling challenges for the realistic application of phage-antibiotic combined therapy.

  15. DISTRIBUTION OF PHAGE TYPES AND TRANSFERABLE DRUG RESISTANCE IN SHIGELLAE

    Directory of Open Access Journals (Sweden)

    K.Badalian

    1981-08-01

    Full Text Available A total of 610 strains of Shigellae isolated from cases of diarrhea in Iran during 1962-73 were studied with respect to their phage type, as well as antibiotic resistance and transferable drug resistance along with serotyping. It was shown that there was some relation between serotypes and phage types but no association could be found between phage types and resistance pattern.

  16. Economics of reducing Campylobacter at different levels within the Belgian poultry meat chain.

    Science.gov (United States)

    Gellynck, Xavier; Messens, Winy; Halet, Dirk; Grijspeerdt, Koen; Hartnett, Emma; Viaene, Jacques

    2008-03-01

    Campylobacter infections pose a serious public health problem in Belgium. Poultry meat is most likely responsible for 40% of human campylobacteriosis cases in Belgium. On a yearly basis, consumption of poultry meat causes at least 22,000 campylobacteriosis cases, with a cost of illness of Euro 10.9 million. Several intervention measures have been proposed in literature, aiming to reduce the contamination of poultry meat and thus lead to significant reductions of human campylobacteriosis cases. This study aimed to evaluate the cost-benefit ratio, i.e., the ratio of reduced costs of illness on intervention costs of various intervention measures. These measures were selected by representatives from the poultry meat sector and experts in the field of poultry science. The selection comprised measures at the farm level (phage therapy), at the processing plant (spraying of carcasses with lactic acid or electrolyzed oxidizing water, crust freezing, or irradiation), and at the consumer level (improving kitchen hygiene and application of home freezing). Among these measures, the decontamination of carcasses with electrolyzed oxidizing water applied in the processing plant was the most efficient (17.66), followed by the use of lactic acid (4.06). In addition, phage therapy generated a positive cost-benefit ratio (2.54). Irradiation indicated the highest efficacy, but its cost-benefit ratio was rather low (0.31). There seems to be less gain by trying to improve food handling in the kitchen. The cost to reach consumers is large, while only a very limited fraction of the consumers is willing to change its behavior. The outcome of this study poses valuable information for future risk-management decisions in Belgium.

  17. Bacteriophages and Phage-Derived Proteins – Application Approaches

    Science.gov (United States)

    Drulis-Kawa, Zuzanna; Majkowska-Skrobek, Grazyna; Maciejewska, Barbara

    2015-01-01

    Currently, the bacterial resistance, especially to most commonly used antibiotics has proved to be a severe therapeutic problem. Nosocomial and community-acquired infections are usually caused by multidrug resistant strains. Therefore, we are forced to develop an alternative or supportive treatment for successful cure of life-threatening infections. The idea of using natural bacterial pathogens such as bacteriophages is already well known. Many papers have been published proving the high antibacterial efficacy of lytic phages tested in animal models as well as in the clinic. Researchers have also investigated the application of non-lytic phages and temperate phages, with promising results. Moreover, the development of molecular biology and novel generation methods of sequencing has opened up new possibilities in the design of engineered phages and recombinant phage-derived proteins. Encouraging performances were noted especially for phage enzymes involved in the first step of viral infection responsible for bacterial envelope degradation, named depolymerases. There are at least five major groups of such enzymes – peptidoglycan hydrolases, endosialidases, endorhamnosidases, alginate lyases and hyaluronate lyases – that have application potential. There is also much interest in proteins encoded by lysis cassette genes (holins, endolysins, spanins) responsible for progeny release during the phage lytic cycle. In this review, we discuss several issues of phage and phage-derived protein application approaches in therapy, diagnostics and biotechnology in general. PMID:25666799

  18. Bacteriophages and phage-derived proteins--application approaches.

    Science.gov (United States)

    Drulis-Kawa, Zuzanna; Majkowska-Skrobek, Grazyna; Maciejewska, Barbara

    2015-01-01

    Currently, the bacterial resistance, especially to most commonly used antibiotics has proved to be a severe therapeutic problem. Nosocomial and community-acquired infections are usually caused by multidrug resistant strains. Therefore, we are forced to develop an alternative or supportive treatment for successful cure of life-threatening infections. The idea of using natural bacterial pathogens such as bacteriophages is already well known. Many papers have been published proving the high antibacterial efficacy of lytic phages tested in animal models as well as in the clinic. Researchers have also investigated the application of non-lytic phages and temperate phages, with promising results. Moreover, the development of molecular biology and novel generation methods of sequencing has opened up new possibilities in the design of engineered phages and recombinant phage-derived proteins. Encouraging performances were noted especially for phage enzymes involved in the first step of viral infection responsible for bacterial envelope degradation, named depolymerases. There are at least five major groups of such enzymes - peptidoglycan hydrolases, endosialidases, endorhamnosidases, alginate lyases and hyaluronate lyases - that have application potential. There is also much interest in proteins encoded by lysis cassette genes (holins, endolysins, spanins) responsible for progeny release during the phage lytic cycle. In this review, we discuss several issues of phage and phage-derived protein application approaches in therapy, diagnostics and biotechnology in general.

  19. Revisiting phage therapy: new applications for old resources.

    Science.gov (United States)

    Nobrega, Franklin L; Costa, Ana Rita; Kluskens, Leon D; Azeredo, Joana

    2015-04-01

    The success of phage therapy is dependent on the development of strategies able to overcome the limitations of bacteriophages as therapeutic agents, the creation of an adequate regulatory framework, the implementation of safety protocols, and acceptance by the general public. Many approaches have been proposed to circumvent phages' intrinsic limitations but none have proved to be completely satisfactory. In this review we present the major hurdles of phage therapy and the solutions proposed to circumvent them. A thorough discussion of the advantages and drawbacks of these solutions is provided and special attention is given to the genetic modification of phages as an achievable strategy to shape bacteriophages to exhibit desirable biological properties.

  20. Isolation and Characterization of Phages Infecting Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Anna Krasowska

    2015-01-01

    Full Text Available Bacteriophages have been suggested as an alternative approach to reduce the amount of pathogens in various applications. Bacteriophages of various specificity and virulence were isolated as a means of controlling food-borne pathogens. We studied the interaction of bacteriophages with Bacillus species, which are very often persistent in industrial applications such as food production due to their antibiotic resistance and spore formation. A comparative study using electron microscopy, PFGE, and SDS-PAGE as well as determination of host range, pH and temperature resistance, adsorption rate, latent time, and phage burst size was performed on three phages of the Myoviridae family and one phage of the Siphoviridae family which infected Bacillus subtilis strains. The phages are morphologically different and characterized by icosahedral heads and contractile (SIOΦ, SUBω, and SPOσ phages or noncontractile (ARπ phage tails. The genomes of SIOΦ and SUBω are composed of 154 kb. The capsid of SIOΦ is composed of four proteins. Bacteriophages SPOσ and ARπ have genome sizes of 25 kbp and 40 kbp, respectively. Both phages as well as SUBω phage have 14 proteins in their capsids. Phages SIOΦ and SPOσ are resistant to high temperatures and to the acid (4.0 and alkaline (9.0 and 10.0 pH.

  1. Learning from Bacteriophages - Advantages and Limitations of Phage and Phage-Encoded Protein Applications

    Science.gov (United States)

    Drulis-Kawa, Zuzanna; Majkowska-Skrobek, Grażyna; Maciejewska, Barbara; Delattre, Anne-Sophie; Lavigne, Rob

    2012-01-01

    The emergence of bacteria resistance to most of the currently available antibiotics has become a critical therapeutic problem. The bacteria causing both hospital and community-acquired infections are most often multidrug resistant. In view of the alarming level of antibiotic resistance between bacterial species and difficulties with treatment, alternative or supportive antibacterial cure has to be developed. The presented review focuses on the major characteristics of bacteriophages and phage-encoded proteins affecting their usefulness as antimicrobial agents. We discuss several issues such as mode of action, pharmacodynamics, pharmacokinetics, resistance and manufacturing aspects of bacteriophages and phage-encoded proteins application. PMID:23305359

  2. Learning from bacteriophages - advantages and limitations of phage and phage-encoded protein applications.

    Science.gov (United States)

    Drulis-Kawa, Zuzanna; Majkowska-Skrobek, Grazyna; Maciejewska, Barbara; Delattre, Anne-Sophie; Lavigne, Rob

    2012-12-01

    The emergence of bacteria resistance to most of the currently available antibiotics has become a critical therapeutic problem. The bacteria causing both hospital and community-acquired infections are most often multidrug resistant. In view of the alarming level of antibiotic resistance between bacterial species and difficulties with treatment, alternative or supportive antibacterial cure has to be developed. The presented review focuses on the major characteristics of bacteriophages and phage-encoded proteins affecting their usefulness as antimicrobial agents. We discuss several issues such as mode of action, pharmacodynamics, pharmacokinetics, resistance and manufacturing aspects of bacteriophages and phage-encoded proteins application.

  3. Campylobacter concisus - a new player in intestinal disease

    Directory of Open Access Journals (Sweden)

    Nadeem Omar Kaakoush

    2012-02-01

    Full Text Available Over the last decade Campylobacter concisus, a highly fastidious member of the Campylobacter genus has been described as an emergent pathogen of the human intestinal tract. Historically, C. concisus was associated with the human oral cavity and has been linked with periodontal lesions, including gingivitis and periodontitis, although currently its role as an oral pathogen remains contentious. Evidence to support the role of C. concisus in acute intestinal disease has come from studies that have detected or isolated C. concisus as sole pathogen in fecal samples from diarrheic patients. C. concisus has also been associated with chronic intestinal disease, its prevalence being significantly higher in children with newly diagnosed Crohn’s disease and adults with ulcerative colitis than in controls. Further C. concisus has been isolated from biopsy specimens of patients with Crohn’s disease. While such studies support the role of C. concisus as an intestinal pathogen, its isolation from healthy individuals, and failure of some studies to show a significant difference in C. concisus prevalence in subjects with diarrhea and healthy controls has raised contention as to its role in intestinal disease. Such findings could argue against the role of C. concisus in intestinal disease, however, the fact that C. concisus strains are genetically diverse raises the possibility that differences exist in their pathogenic potential. Evidence to support this view comes from studies showing strain specific differences in the ability of C. concisus to attach to and invade cells and produce virulence factors, including toxins and hemolytic phospholipase A. Further, sequencing of the genome of a C. concisus strain isolated from a child with Crohn’s disease (UNSWCD and comparison of this with the only other fully sequenced strain (BAA-1457 would suggest that major differences exist in the genetic make-up of this species which could explain different outcomes of C

  4. Microbiological criteria for Campylobacter in broiler carcasses in Italy: a possible approach to derive them.

    Science.gov (United States)

    Comin, Damiano; Valero, Antonio; Manfreda, Gerardo; García-Gimeno, Rosa Maria; Paiusco, Antonella; De Medici, Dario; Terza, Paolo; Ferrarini, Stefano; De Cesare, Alessandra

    2014-08-01

    The aim of this paper was to provide suitable microbiological criteria (MC) for Campylobacter in broiler carcasses and a sampling plan to verify compliance with such criteria. Data were gathered in the presence and concentration of Campylobacter in broiler carcasses collected in three different Italian slaughterhouses, labelled as A, B and C. The sampling plan to be validated in each slaughterhouse included the analysis of three different carcasses collected immediately after chilling from 30 different lots, for a total of 90 samples per slaughterhouse. The number of positive samples containing above 100 CFU/g and above 1,000 CFU/g throughout the 30 tested lots was determined to estimate between-lot variability. Based on this information, the performance of four MC was evaluated for lot compliance: i) n=3; c=0; m=100 CFU/g; ii) n=3; c=0; m=1,000 CFU/g; iii) n=3; c=1; m=1,000 CFU/g and iv) n=3; c=2; m=1,000 CFU/g. Positive Campylobacter samples were found in 60% of the lots tested in slaughterhouses A and C and in 73.3% of lots from slaughterhouse B. The differences among the three slaughterhouses in the mean Campylobacter levels found in positive samples were not significant and were used to evaluate the performance of the MC. The level of lot compliance to different MC was calculated and for the most stringent one (n=3; c=0; m=100 CFU/g) was 40% at slaughterhouses A and C but only 26.7% at slaughterhouse B. The results of this study show an alternative approach to establish MC for Campylobacter in broilers. According to (1) Campylobacter prevalence and concentration in Italy, (2) applied experimental plan and (3) selected slaughterhouses, the number of compliant lots to the suggested MC ranged between 26.7 and 100%. The selection of the fit for purpose MC is a risk manager decision, based on a reasonable balance between public health and cost for poultry industries.

  5. Prevalence of Salmonella and Campylobacter in beef cattle from transport to slaughter.

    Science.gov (United States)

    Beach, John C; Murano, Elsa A; Acuff, Gary R

    2002-11-01

    The objective of this study was to evaluate the effect of typical production practices during the transport of cattle on the resulting incidence of Salmonella and Campylobacter in the feces, on the hides, and on the carcasses of these cattle and in the environment (trucks, holding pens, and knock boxes). Various factors were evaluated, including the type of animal (feedlot cattle vs. adult pasture cattle), the breed of cattle, the body condition of the animal, the age of the animal, the time of feed and water withdrawal, the contamination level of the transport vehicle at the feedlot or farm, the transport time, the time cattle were held in the holding pen at the plant, and the contamination level of the holding pen. Four groups of each type of animal were sampled on different days. Samples were collected from cattle prior to transport and after transport (rectal and hide swabs) as well as from the carcasses of these cattle. Pre- and posttransit samples were also taken from the transport vehicle and from the holding pen and knock box at the slaughter facility. For feedlot cattle, fecal shedding stayed fairly constant for both organisms before and after transport (3 to 5% for Salmonella and 64 to 68% for Campylobacter). However, the shedding rate for adult cattle increased from 1 to 21% for Salmonella but stayed constant for Campylobacter (6 to 7%). Contamination of hides with Salmonella increased for both animal types from a level of 18 to 20% to a level 50 to 56%. For Campylobacter, the contamination level decreased from 25 to 13% for feedlot cattle but remained unchanged for adult animals (1 to 2%). Nineteen percent of feedlot cattle carcasses and 54% of adult cattle carcasses tested positive for Salmonella, while only2% of feedlot cattle carcasses and none of the adult cattle carcasses tested positive for Campylobacter. Thus, for feedlot cattle, the factors considered in this study did not affect the shedding of either organism but did affect the contamination

  6. Campylobacter jejuni and Campylobacter coli in wild birds on Danish livestock farms

    DEFF Research Database (Denmark)

    Hald, Birthe; Skov, Marianne Nielsine; Nielsen, Eva Møller;

    2016-01-01

    Background: Reducing the occurrence of campylobacteriosis is a food safety issue of high priority, as in recent years it has been the most commonly reported zoonosis in the EU. Livestock farms are of particular interest, since cattle, swine and poultry are common reservoirs of Campylobacter spp. ...

  7. Computer-assisted analysis and epidemiological value of genotyping methods for Campylobacter jejuni and Campylobacter coli

    NARCIS (Netherlands)

    Boer, P. de; Duim, B.; Rigter, A.; Plas, J. van der; Jacobs-Reitsma, W.F.; Wagenaar, J.A.

    2000-01-01

    For epidemiological tracing of the thermotolerant Campylobacter species C. jejuni and C. coli, reliable and highly discriminatory typing techniques are necessary. In this study the genotyping techniques of flagellin typing (flaA typing), pulsed-field gel electrophoresis (PFGE), automated ribotyping,

  8. Campylobacter jejuni and Campylobacter coli in wild birds on Danish livestock farms

    DEFF Research Database (Denmark)

    Hald, Birthe; Skov, Marianne Nielsine; Nielsen, Eva Møller

    2016-01-01

    Background: Reducing the occurrence of campylobacteriosis is a food safety issue of high priority, as in recent years it has been the most commonly reported zoonosis in the EU. Livestock farms are of particular interest, since cattle, swine and poultry are common reservoirs of Campylobacter spp...

  9. Tight junction changes in epithelial cells by Campylobacter jejuni and non-jejuni Campylobacter species

    DEFF Research Database (Denmark)

    Bücker, Roland; Nielsen, Hans Linde; Krüg, S

    Campylobacter concisus infections of the gastrointestinal tract can be accompanied by diarrhea and inflammation, whereas colonization of the human oral cavity might have a commensal nature. We focus on the pathophysiology of C. concisus and the effects of different clinical oral and fecal C...

  10. Quantifying potential sources of surface water contamination with Campylobacter jejuni and Campylobacter coli

    NARCIS (Netherlands)

    Mughini-Gras, Lapo; Penny, Christian; Ragimbeau, Catherine; Schets, Franciska M.; Blaak, Hetty; Duim, Birgitta; Wagenaar, Jaap A.; Boer, de Albert; Cauchie, Henry-Michel; Mossong, Joel; Pelt, Van Wilfrid

    2016-01-01

    Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water

  11. Computer-assisted analysis and epidemiological value of genotyping methods for Campylobacter jejuni and Campylobacter coli

    NARCIS (Netherlands)

    Boer, P. de; Duim, B.; Rigter, A.; Plas, J. van der; Jacobs-Reitsma, W.F.; Wagenaar, J.A.

    2000-01-01

    For epidemiological tracing of the thermotolerant Campylobacter species C. jejuni and C. coli, reliable and highly discriminatory typing techniques are necessary. In this study the genotyping techniques of flagellin typing (flaA typing), pulsed-field gel electrophoresis (PFGE), automated ribotyping,

  12. Higher resistance of Campylobacter coli compared to Campylobacter jejuni at chicken slaughterhouse.

    Science.gov (United States)

    Torralbo, Alicia; Borge, Carmen; García-Bocanegra, Ignacio; Méric, Guillaume; Perea, Anselmo; Carbonero, Alfonso

    2015-04-01

    In order to compare the prevalence of Campylobacter coli and Campylobacter jejuni during the processing of broilers at slaughterhouse a total of 848 samples were analyzed during 2012 in southern Spain. Four hundred and seventy six samples were collected from cloaca, carcass surfaces and quartered carcasses. Moreover, 372 environmental swabs from equipment and scalding water were collected. Minimum inhibitory concentration (MIC) to ciprofloxacin, erythromycin, streptomycin, tetracycline and gentamicin was determined for isolates from chicken meat. The general prevalence of Campylobacter was 68.8% (40.2% of C. coli and 28.5% of C. jejuni). The relative prevalence of C. coli increased from loading dock area (41.5%) to packing area (64.6%). In contrast, the relative prevalence of C. jejuni decreased from 58.5% to 35.4%. These differences between species from initial to final area were significant (p=0.02). The highest antimicrobial resistance for C. jejuni and C. coli was detected to tetracycline (100%) and ciprofloxacin (100%), respectively. Campylobacter coli showed an antimicrobial resistance significantly higher than C. jejuni to streptomycin (p=0.002) and erythromycin (p<0.0001). Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Campylobacter jejuni: exposure assessment and hazard characterization : growth, survival and infectivity of Campylobacter jejuni

    NARCIS (Netherlands)

    Verhoeff-Bakkenes, L.

    2012-01-01

    Campylobacter jejuni, a small, curved or spirally shaped highly motile microorganism, is identified as a major cause of bacterial gastroenteritis throughout the world. Serious complications such as the Guillain-Barré syndrome and reactive arthritis might occasionally follow infection. In this

  14. Quantifying potential sources of surface water contamination with Campylobacter jejuni and Campylobacter coli

    NARCIS (Netherlands)

    Mughini-Gras, Lapo; Penny, Christian; Ragimbeau, Catherine; Schets, Franciska M.; Blaak, Hetty; Duim, Birgitta; Wagenaar, Jaap A.; Boer, de Albert; Cauchie, Henry-Michel; Mossong, Joel; Pelt, Van Wilfrid

    2016-01-01

    Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water contam

  15. Toename Salmonella en Campylobacter bij slacht : Campylobacter in pluimveesector constant, Salmonella sterk gedaald

    NARCIS (Netherlands)

    Fels-Klerx, van der H.J.; Asselt, van E.D.

    2009-01-01

    In vrijwel alle schakels van de pluimveevleesketen is de besmetting met Salmonella in de periode van 2002 tot 2005 gedaald. Campylobacter werd in diezelfde periode juist vaker in slachthuizen aangetroffen. Tijdens het slachten nam de besmetting met beide pathogenen toe, voor Salmonella gold dit voor

  16. Campylobacter jejuni: exposure assessment and hazard characterization : growth, survival and infectivity of Campylobacter jejuni

    NARCIS (Netherlands)

    Verhoeff-Bakkenes, L.

    2012-01-01

    Campylobacter jejuni, a small, curved or spirally shaped highly motile microorganism, is identified as a major cause of bacterial gastroenteritis throughout the world. Serious complications such as the Guillain-Barré syndrome and reactive arthritis might occasionally follow infection. In this

  17. Development of a selective agar plate for the detection of Campylobacter spp. in fresh produce.

    Science.gov (United States)

    Yoo, Jin-Hee; Choi, Na-Young; Bae, Young-Min; Lee, Jung-Su; Lee, Sun-Young

    2014-10-17

    This study was conducted to develop a selective medium for the detection of Campylobacter spp. in fresh produce. Campylobacter spp. (n=4), non-Campylobacter (showing positive results on Campylobacter selective agar) strains (n=49) isolated from fresh produce, indicator bacteria (n=13), and spoilage bacteria isolated from fresh produce (n=15) were plated on four Campylobacter selective media. Bolton agar and modified charcoal cefoperazone deoxycholate agar (mCCDA) exhibited higher sensitivity for Campylobacter spp. than did Preston agar and Hunt agar, although certain non-Campylobacter strains isolated from fresh produce by using a selective agar isolation method, were still able to grow on Bolton agar and mCCDA. To inhibit the growth of non-Campylobacter strains, Bolton agar and mCCDA were supplemented with 5 antibiotics (rifampicin, polymyxin B, sodium metabisulfite, sodium pyruvate, ferrous sulfate) and the growth of Campylobacter spp. (n=7) and non-Campylobacter strains (n=44) was evaluated. Although Bolton agar supplemented with rifampicin (BR agar) exhibited a higher selectivity for Campylobacter spp. than did mCCDA supplemented with antibiotics, certain non-Campylobacter strains were still able to grow on BR agar (18.8%). When BR agar with various concentrations of sulfamethoxazole-trimethoprim were tested with Campylobacter spp. (n=8) and non-Campylobacter (n=7), sulfamethoxazole-trimethoprim was inhibitory against 3 of 7 non-Campylobacter strains. Finally, we validated the use of BR agar containing 50mg/L sulfamethoxazole (BRS agar) or 0.5mg/L ciprofloxacin (BRCS agar) and other selective agars for the detection of Campylobacter spp. in chicken and fresh produce. All chicken samples were positive for Campylobacter spp. when tested on mCCDA, BR agar, and BRS agar. In fresh produce samples, BRS agar exhibited the highest selectivity for Campylobacter spp., demonstrating its suitability for the detection of Campylobacter spp. in fresh produce.

  18. Exploring PFGE for Detecting Large Plasmids in Campylobacter jejuni and Campylobacter coli Isolated from Various Retail Meats

    Directory of Open Access Journals (Sweden)

    Daya Marasini

    2014-10-01

    Full Text Available Campylobacter spp. is one of the most prevalent bacterial pathogens in retail meat, particularly poultry, and is a leading cause of diarrhea in humans. Studies related to Campylobacter large plasmids are limited in the literature possibly due to difficulty in isolating them using available alkaline lysis methods. The objectives of this study were to determine the prevalence of plasmids, particularly large ones, in Campylobacter spp. isolated from various Oklahoma retail meats, and to explore PFGE (Pulsed Field Gel Electrophoresis as a tool in facilitating the detection of these plasmids. One hundred and eighty nine strains (94 Campylobacter jejuni and 95 Campylobacter coli were screened for the presence of plasmids using both alkaline lysis and PFGE. Plasmids were detected in 119/189 (63% using both methods. Most of the plasmids detected by alkaline lysis were smaller than 90 kb and only three were larger than 90 kb. Plasmids over 70 kb in size were detected in 33 more strains by PFGE of which 11 strains contained larger than 90 kb plasmids. Plasmids were more prevalent in Campylobacter coli (73.5% than in Campylobacter jejuni (52%. BglII restriction analysis of plasmids isolated from 102 isolates revealed 42 different restriction patterns. In conclusion, PFGE was able to detect large plasmids up to 180 Kb in Campylobacter spp. which might have been missed if the alkaline lysis method was solely used. Campylobacter spp. isolated from retail meats harbor a diverse population of plasmids with variable sizes. To our knowledge, this is the first study to use PFGE to detect large plasmids in Campylobacter.

  19. Trisodium phosphate and sodium hypochlorite are more effective as antimicrobials against Campylobacter and Salmonella on duck as compared to chicken meat.

    Science.gov (United States)

    Sarjit, Amreeta; Dykes, Gary A

    2015-06-16

    Little work has been reported on the use of commercial antimicrobials against foodborne pathogens on duck meat. We investigated the effectiveness of trisodium phosphate (TSP) and sodium hypochlorite (SH) as antimicrobial treatments against Campylobacter and Salmonella on duck meat under simulated commercial water chilling conditions. The results were compared to the same treatments on well-studied chicken meat. A six strain Campylobacter or Salmonella cocktail was inoculated (5 ml) at two dilution levels (10(4) and 10(8) cfu/ml) onto 25 g duck or chicken meat with skin and allowed to attach for 10 min. The meat was exposed to three concentrations of pH adjusted TSP (8, 10 and 12% (w/v), pH 11.5) or SH (40, 50 and 60 ppm, pH 5.5) in 30 ml water under simulated spin chiller conditions (4 °C, agitation) for 10 min. In a parallel experiment the meat was placed in the antimicrobial treatments before inoculation and bacterial cocktails were added to the meat after the antimicrobial solution was removed while all other parameters were maintained. Untreated controls and controls using water were included in all experiments. Bacterial numbers were determined on Campylobacter blood-free selective agar and Mueller Hinton agar or xylose deoxycholate agar and tryptone soya agar using the thin agar layer method for Campylobacter and Salmonella, respectively. All TSP concentrations significantly (pCampylobacter (~1.2-6.4 log cfu/cm(2)) and Salmonella (~0.4-6.6 log cfu/cm(2)) on both duck and chicken meat. On duck meat, numbers of Campylobacter were less than the limit of detection at higher concentrations of TSP and numbers of Salmonella were less than the limit of detection at all concentrations of TSP except one. On chicken meat, numbers of Campylobacter and Salmonella were less than the limit of detection only at the lower inoculum level and higher TSP concentrations. By contrast only some of the concentrations of SH significantly (pCampylobacter and Salmonella (~0.2-1.5 log

  20. Proteomic and genomic analysis reveals novel Campylobacter jejuni outer membrane proteins and potential heterogeneity

    Directory of Open Access Journals (Sweden)

    Eleanor Watson

    2014-09-01

    Full Text Available Gram-negative bacterial outer membrane proteins play important roles in the interaction of bacteria with their environment including nutrient acquisition, adhesion and invasion, and antibiotic resistance. In this study we identified 47 proteins within the Sarkosyl-insoluble fraction of Campylobacter jejuni 81-176, using LC–ESI-MS/MS. Comparative analysis of outer membrane protein sequences was visualised to reveal protein distribution within a panel of Campylobacter spp., identifying several C. jejuni-specific proteins. Smith–Waterman analyses of C. jejuni homologues revealed high sequence conservation amongst a number of hypothetical proteins, sequence heterogeneity of other proteins and several proteins which are absent in a proportion of strains.

  1. Campylobacter-Associated Hemolytic Uremic Syndrome Associated with Pulmonary-Renal Syndrome.

    Science.gov (United States)

    Bowen, Emily Elizabeth; Hangartner, Robert; Macdougall, Iain

    2016-03-01

    Common causes of pulmonary-renal syndrome include anti-glomerular basement membrane (anti-GBM) disease anti-neutrophil cytoplasmic antibody (ANCA) positive vasculitis, and systemic lupus erythematosus. We describe a case of life-threatening pulmonary hemorrhage associated with Campylobacter hemolytic uremic syndrome (HUS), which we believe is a new disease entity. We hypothesize that the cause of this pulmonary-renal syndrome was an immunological reaction to Campylobacter; and that the initiation of high-dose steroids was responsible for the rapid reversal of the patient's pulmonary and renal impairment. The aim of this article is to raise awareness of this unusual cause of a pulmonary-renal syndrome, guiding physicians to recognize it as a potential complication, and to consider high-dose steroids in managing the condition.

  2. Complete Genome Sequences of Four Novel Escherichia coli Bacteriophages Belonging to New Phage Groups

    DEFF Research Database (Denmark)

    Carstens, Alexander B; Kot, Witold; Hansen, Lars H

    2015-01-01

    Here, we describe the sequencing and genome annotations of a set of four Escherichia coli bacteriophages (phages) belonging to newly discovered groups previously consisting of only a single phage and thus expand our knowledge of these phage groups.......Here, we describe the sequencing and genome annotations of a set of four Escherichia coli bacteriophages (phages) belonging to newly discovered groups previously consisting of only a single phage and thus expand our knowledge of these phage groups....

  3. Facing antibiotic resistance: Staphylococcus aureus phages as a medical tool.

    Science.gov (United States)

    Kaźmierczak, Zuzanna; Górski, Andrzej; Dąbrowska, Krystyna

    2014-07-01

    Staphylococcus aureus is a common and often virulent pathogen in humans. This bacterium is widespread, being present on the skin and in the nose of healthy people. Staphylococcus aureus can cause infections with severe outcomes ranging from pustules to sepsis and death. The introduction of antibiotics led to a general belief that the problem of bacterial infections would be solved. Nonetheless, pathogens including staphylococci have evolved mechanisms of drug resistance. Among current attempts to address this problem, phage therapy offers a promising alternative to combat staphylococcal infections. Here, we present an overview of current knowledge on staphylococcal infections and bacteriophages able to kill Staphylococcus, including experimental studies and available data on their clinical use.

  4. Facing Antibiotic Resistance: Staphylococcus aureus Phages as a Medical Tool

    Directory of Open Access Journals (Sweden)

    Zuzanna Kaźmierczak

    2014-07-01

    Full Text Available Staphylococcus aureus is a common and often virulent pathogen in humans. This bacterium is widespread, being present on the skin and in the nose of healthy people. Staphylococcus aureus can cause infections with severe outcomes ranging from pustules to sepsis and death. The introduction of antibiotics led to a general belief that the problem of bacterial infections would be solved. Nonetheless, pathogens including staphylococci have evolved mechanisms of drug resistance. Among current attempts to address this problem, phage therapy offers a promising alternative to combat staphylococcal infections. Here, we present an overview of current knowledge on staphylococcal infections and bacteriophages able to kill Staphylococcus, including experimental studies and available data on their clinical use.

  5. CAMPYLOBACTER LARI AND CAMPYLOBACTER UPSALIENSIS -NEW ENTITIES IN THE CAMPYLOBACTERIOSES ETIOLOGY

    Directory of Open Access Journals (Sweden)

    Gordana Tasić

    2002-03-01

    Full Text Available The campylobacterioses are diseases occurring in people and animals and they are caused by the bacteria of the Campylobacter gender. The Campylobacter gender today comprises 15 species though the status of some of them has not been precisely defined yet. Most of them are isolated from the material of human origin; but with the exception of C. jejuni and C. coli, the other species are of less importance for human medicine. Beside C. jejuni and C. coli today there are other species of the Campylobacters acknowledged as the human pathogens in immune-compromised persons, homosexuals and in overall human population. The gender representative, C. fetus subsp. fetus, has been confirmed as the main cause of abortion in sheep and cows; it is rarely recognized in human pathology but once it is found it is usually in immunedeficient persons. Since 1984 in scientific literature in the world new entities in the campylobacterioses have been described. Campylobacter lari is a potentially pathogenic species that is rarely described. The pathogenic potential of this bacteria for people is for the first time confirmed in 1984 in the description of a case of fatal bacterioremia caused bu C. lari in some immune-compromised patients. C. lari is described as the cause of enteritis, gastroenteritis, appendicitis and purulent pleuritis. The gastroenteritis epidemic caused by C. lari was described in 1985; itemerged in an area inhabited by sea-gulls. C. upsaliensis is today confirmed as a Campylobacter species causing diarrhea and bacteremia in healthy and immune-compromised persons. This microorganism can be even more frequent cause of gastroenteritis than shown by official data.

  6. Replicating phages in the epidermal mucosa of the eel (Anguilla anguilla

    Directory of Open Access Journals (Sweden)

    Miguel eCarda Diéguez

    2015-01-01

    Full Text Available In this work, we used the eel (Anguilla anguilla as an animal model to test the hypothesis of Barr et al. about the putative role of the epidermal mucosa as a phage enrichment layer. To this end, we analyzed the microbial content of the skin mucus of wild and farmed eels by using a metagenomic approach. We found a great abundance of replicating phages (concatemers in all the samples. They were assembled in four complete genomes of three Myovirus and one Podovirus. We also found evidences that ΦKZ and Podovirus phages could be part of the resident microbiota associated to the eel mucosal surface and persist on them over the time. Moreover, the viral abundance estimated by epiflorescent counts and by metagenomic recruitment from eel mucosa was higher than that of the surrounding water. Taken together, our results support the hypothesis that claims a possible role of phages in the animal mucus as agents controlling bacterial populations, including pathogenic species, providing a kind of innate immunity.

  7. Characterization of Bacillus phage-K2 isolated from chungkookjang, a fermented soybean foodstuff.

    Science.gov (United States)

    Kim, Eun Ju; Hong, Jeong Won; Yun, Na-Rae; Lee, Young Nam

    2011-01-01

    An investigation of a virulent Bacillus phage-K2 (named Bp-K2) isolated from chungkookjang (a fermented soybean foodstuff) was made. Bp-K2 differed in infectivity against a number of Bacillus subtilis strains including starter strains of chungkookjang and natto, being more infectious to Bacillus strains isolated from the chungkookjang, but much less active against a natto strain. Bp-K2 is a small DNA phage whose genome size is about 21 kb. Bp-K2 is a tailed bacteriophage with an isometric icosahedral head (50 nm long on the lateral side, 80 nm wide), a long contractile sheath (85-90 nm × 28 nm), a thin tail fiber (80-85 nm long, 10 nm wide), and a basal plate (29 nm long, 47 nm wide) with a number of spikes, but no collar. The details of the structures of Bp-K2 differ from natto phage ϕBN100 as well as other known Bacillus phages such as SPO1-like or ϕ 29-like viruses. These data suggest that Bp-K2 would be a new member of the Myoviridae family of Bacillus bacteriophages.

  8. Optimality models of phage life history and parallels in disease evolution.

    Science.gov (United States)

    Bull, J J

    2006-08-21

    Optimality models constitute one of the simplest approaches to understanding phenotypic evolution. Yet they have shortcomings that are not easily evaluated in most organisms. Most importantly, the genetic basis of phenotype evolution is almost never understood, and phenotypic selection experiments are rarely possible. Both limitations can be overcome with bacteriophages. However, phages have such elementary life histories that few phenotypes seem appropriate for optimality approaches. Here we develop optimality models of two phage life history traits, lysis time and host range. The lysis time models show that the optimum is less sensitive to differences in host density than suggested by earlier analytical work. Host range evolution is approached from the perspective of whether the virus should avoid particular hosts, and the results match optimal foraging theory: there is an optimal "diet" in which host types are either strictly included or excluded, depending on their infection qualities. Experimental tests of both models are feasible, and phages provide concrete illustrations of many ways that optimality models can guide understanding and explanation. Phage genetic systems already support the perspective that lysis time and host range can evolve readily and evolve without greatly affecting other traits, one of the main tenets of optimality theory. The models can be extended to more general properties of infection, such as the evolution of virulence and tissue tropism.

  9. Phage endolysins with broad antimicrobial activity against Enterococcus faecalis clinical strains.

    Science.gov (United States)

    Proença, Daniela; Fernandes, Sofia; Leandro, Clara; Silva, Filipa Antunes; Santos, Sofia; Lopes, Fátima; Mato, Rosario; Cavaco-Silva, Patrícia; Pimentel, Madalena; São-José, Carlos

    2012-06-01

    Increasing antibiotic resistance of bacterial pathogens has drawn the attention to the potential use of bacteriophage endolysins as alternative antibacterial agents. Here we have identified, characterized, and studied the lytic potential of two endolysins, Lys168 and Lys170, from phages infecting Enterococcus faecalis. Lys168 and Lys170 belong to the cysteine, histidine-dependent amidohydrolases/peptidases (CHAP) and amidase-2 protein families, respectively. Lys168 is quite a unique enterococcal phage endolysin. It shares 95% amino acidic identity with the endolysin of Staphylococcus aureus phage SAP6, which in turn is distantly related to all known CHAP endolysins of S. aureus phages. Lys170 seems to be a natural chimera assembling catalytic and cell-wall-binding domains of different origin. Both endolysins showed a clear preference to act against E. faecalis and they were able to lyse a high proportion of clinical isolates of this species. Specifically, Lys168 and Lys170 lysed more than 70% and 90% of the tested isolates, respectively, which included a panel of diverse and typed strains representative of highly prevalent clonal complexes. Lys170 was active against all tested E. faecalis VRE strains. The quasi specificity toward E. faecalis is discussed considering the nature of the enzymes' functional domains and the structure of the cell wall peptidoglycan.

  10. Occurrence of Campylobacter spp. in water in Northern Ireland: implications for public health.

    Science.gov (United States)

    Moore, J E; Caldwell, P S; Millar, B C; Murphy, P G

    2001-11-01

    The occurrence of Campylobacter spp was examined in a variety of waters in Northern Ireland. Conventional cultural techniques were employed with 768 water specimens, including drinking waters (tap, spring, bore hole and bottled) and recreational waters (swimming pool, lough, river and sea). Positive waters included 1/11 (9.1%) drinking water from untreated well water, as well as 5/12 (41.7%) untreated surface waters from loughs and 7/8 (87.5%) untreated river waters. Overall, untreated surface waters may represent a source of contamination with Campylobacter spp. in Northern Ireland, where they have a recreational involvement or are used as a drinking source by man or agricultural livestock. Therefore waterborne campylobacteriosis should be considered in patients presenting with acute enteritis and a history of participation in water sports/activities. As faecal coliform organisms have been previously shown to be poor markers of water quality, especially for Campylobacter spp, new criteria should be established to assess the risk of this infection and to evaluate and monitor the quality of water used for recreational purposes.

  11. Predominant Campylobacter jejuni sequence types persist in Finnish chicken production.

    Directory of Open Access Journals (Sweden)

    Ann-Katrin Llarena

    Full Text Available Consumption and handling of chicken meat are well-known risk factors for acquiring campylobacteriosis. This study aimed to describe the Campylobacter jejuni population in Finnish chickens and to investigate the distribution of C. jejuni genotypes on Finnish chicken farms over a period of several years. We included 89.8% of the total C. jejuni population recovered in Finnish poultry during 2004, 2006, 2007, 2008, and 2012 and used multilocus sequence typing (MLST and pulsed-field gel electrophoresis (PFGE to characterize the 380 isolates. The typing data was combined with isolate information on collection-time and farm of origin. The C. jejuni prevalence in chicken slaughter batches was low (mean 3.0%, CI95% [1.8%, 4.2%], and approximately a quarter of Finnish chicken farms delivered at least one positive chicken batch yearly. In general, the C. jejuni population was diverse as represented by a total of 63 sequence types (ST, but certain predominant MLST lineages were identified. ST-45 clonal complex (CC accounted for 53% of the isolates while ST-21 CC and ST-677 CC covered 11% and 9% of the isolates, respectively. Less than half of the Campylobacter positive farms (40.3% delivered C. jejuni-contaminated batches in multiple years, but the genotypes (ST and PFGE types generally varied from year to year. Therefore, no evidence for a persistent C. jejuni source for the colonization of Finnish chickens emerged. Finnish chicken farms are infrequently contaminated with C. jejuni compared to other European Union (EU countries, making Finland a valuable model for further epidemiological studies of the C. jejuni in poultry flocks.

  12. Characterization of Five Podoviridae Phages Infecting Citrobacter freundii.

    Science.gov (United States)

    Hamdi, Sana; Rousseau, Geneviève M; Labrie, Simon J; Kourda, Rim S; Tremblay, Denise M; Moineau, Sylvain; Slama, Karim B

    2016-01-01

    Citrobacter freundii causes opportunistic infections in humans and animals, which are becoming difficult to treat due to increased antibiotic resistance. The aim of this study was to explore phages as potential antimicrobial agents against this opportunistic pathogen. We isolated and characterized five new virulent phages, SH1, SH2, SH3, SH4, and SH5 from sewage samples in Tunisia. Morphological and genomic analyses revealed that the five C. freundii phages belong to the Caudovirales order, Podoviridae family, and Autographivirinae subfamily. Their linear double-stranded DNA genomes range from 39,158 to 39,832 bp and are terminally redundant with direct repeats between 183 and 242 bp. The five genomes share the same organization as coliphage T7. Based on genomic comparisons and on the phylogeny of the DNA polymerases, we assigned the five phages to the T7virus genus but separated them into two different groups. Phages SH1 and SH2 are very similar to previously characterized phages phiYeO3-12 and phiSG-JL2, infecting, respectively, Yersinia enterocolitica and Salmonella enterica, as well as sharing more than 80% identity with most genes of coliphage T7. Phages SH3, SH4, and SH5 are very similar to phages K1F and Dev2, infecting, respectively, Escherichia coli and Cronobacter turicensis. Several structural proteins of phages SH1, SH3, and SH4 were detected by mass spectrometry. The five phages were also stable from pH 5 to 10. No genes coding for known virulence factors or integrases were found, suggesting that the five isolated phages could be good candidates for therapeutic applications to prevent or treat C. freundii infections. In addition, this study increases our knowledge about the evolutionary relationships within the T7virus genus.

  13. Arsenic resistance and prevalence of arsenic resistance genes in Campylobacter jejuni and Campylobacter coli isolated from retail meats.

    Science.gov (United States)

    Noormohamed, Aneesa; Fakhr, Mohamed K

    2013-08-07

    Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli) isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4-2,048 μg/mL), roxarsone (4-2048 μg/mL), arsenate (16-8,192 μg/mL) and arsenite (4-2,048 μg/mL). A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli) were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB) by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512-2,048 μg/mL), roxarsone (512-2,048 μg/mL), and arsenate (128-1,024 μg/mL), but at lower concentrations for arsenite (4-16 μg/mL). Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3), 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR). The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates.

  14. Arsenic Resistance and Prevalence of Arsenic Resistance Genes in Campylobacter jejuni and Campylobacter coli Isolated from Retail Meats

    Directory of Open Access Journals (Sweden)

    Mohamed K. Fakhr

    2013-08-01

    Full Text Available Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4–2,048 μg/mL, roxarsone (4–2048 μg/mL, arsenate (16–8,192 μg/mL and arsenite (4–2,048 μg/mL. A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512–2,048 μg/mL, roxarsone (512–2,048 μg/mL, and arsenate (128–1,024 μg/mL, but at lower concentrations for arsenite (4–16 μg/mL. Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3, 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR. The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates.

  15. Arsenic Resistance and Prevalence of Arsenic Resistance Genes in Campylobacter jejuni and Campylobacter coli Isolated from Retail Meats

    Science.gov (United States)

    Noormohamed, Aneesa; Fakhr, Mohamed K.

    2013-01-01

    Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli) isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4–2,048 μg/mL), roxarsone (4–2048 μg/mL), arsenate (16–8,192 μg/mL) and arsenite (4–2,048 μg/mL). A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli) were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB) by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512–2,048 μg/mL), roxarsone (512–2,048 μg/mL), and arsenate (128–1,024 μg/mL), but at lower concentrations for arsenite (4–16 μg/mL). Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3), 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR). The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates. PMID:23965921

  16. Climate variability and campylobacter infection: an international study

    Science.gov (United States)

    Sari Kovats, R.; Edwards, Sally J.; Charron, Dominique; Cowden, John; D'Souza, Rennie M.; Ebi, Kristie L.; Gauci, Charmaine; Gerner-Smidt, Peter; Hajat, Shakoor; Hales, Simon; Hernández Pezzi, Gloria; Kriz, Bohumir; Kutsar, Kuulo; McKeown, Paul; Mellou, Kassiani; Menne, Bettina; O'Brien, Sarah; Pelt, Wilfrid; Schmid, Hans

    2005-03-01

    Campylobacter is among the most important agents of enteritis in developed countries. We have described the potential environmental determinants of the seasonal pattern of infection with campylobacter in Europe, Canada, Australia and New Zealand. Specifically, we investigated the role of climate variability on laboratory-confirmed cases of campylobacter infection from 15 populations. Regression analysis was used to quantify the associations between timing of seasonal peaks in infection in space and time. The short-term association between weekly weather and cases was also investigated using Poisson regression adapted for time series data. All countries in our study showed a distinct seasonality in campylobacter transmission, with many, but not all, populations showing a peak in spring. Countries with milder winters have peaks of infection earlier in the year. The timing of the peak of infection is weakly associated with high temperatures 3 months previously. Weekly variation in campylobacter infection in one region of the UK appeared to be little affected by short-term changes in weather patterns. The geographical variation in the timing of the seasonal peak suggests that climate may be a contributing factor to campylobacter transmission. The main driver of seasonality of campylobacter remains elusive and underscores the need to identify the major serotypes and routes of transmission for this disease.

  17. Presence of Campylobacter spp. in refrigerated chicken cuts

    Directory of Open Access Journals (Sweden)

    Juliane Alves

    2013-12-01

    Full Text Available Campylobacter spp. is a common cause of bacterial food-borne illness. Birds, especially poultry are primary reservoirs of C. jejuni. The aim of this study was to evaluate the occurrence of Campylobacter spp. in chicken cuts purchased in supermarkets of Londrina, Parana. A total of 50 samples of chicken cuts, such as breasts, thighs and drumsticks were analyzed. The confirmation of the presence of Campylobacter spp. was performed by identifying the suspected colonies on the selective medium using the polymerase chain reaction. Of the 50 samples analyzed, 28 (56% were positive for Campylobacter spp. Chicken meat, as observed in this study, is a possible source of Campylobacter transmission to humans. This study alerts for the importance to analyze the occurrence of Campylobacter in chicken meat, due to the significant number of positive samples observed and no available epidemiological data in Brazil. The correct orientation about handling and cooking of chicken meat is also necessary to prevent human infection by Campylobacter spp.

  18. Characterization and Testing the Efficiency of Acinetobacter baumannii Phage vB-GEC_Ab-M-G7 as an Antibacterial Agent

    Directory of Open Access Journals (Sweden)

    Ia Kusradze

    2016-10-01

    Full Text Available Acinetobacter baumannii is a gram-negative, non-motile bacterium that, due to its multidrug resistance, has become a major nosocomial pathogen .The increasing number of multidrug resistant (MDR strains has renewed interest in phage therapy. The aim of our study was to assess the effectiveness of phage administration in Acinetobacter baumannii wound infections in an animal model to demonstrate phage therapy as non-toxic, safe and alternative antibacterial remedy. Using classical methods for the study of bacteriophage properties, we characterized phage vB-GEC_Ab-M-G7 as a dsDNA myovirus with a 90kb genome size. Important characteristics of vB-GEC_Ab-M-G7include a short latent period and large burst size, wide host range, resistance to chloroform and thermal and pH stability. In a rat wound model, phage application effectively decreased the number of bacteria isolated from the wounds of successfully treated animals. This study highlights the effectiveness of the phage therapy and provides further insight into treating infections caused by MDR strains using phage administration.

  19. Improved Binding Activity of Antibodies against Major Histocompatibility Complex Class I Chain-Related Gene A by Phage Display Technology for Cancer-Targeted Therapy

    Directory of Open Access Journals (Sweden)

    Achara Phumyen

    2012-01-01

    Full Text Available Major histocompatibility complex class I chain-related gene A (MICA is an NKG2D ligand that is over-expressed under cellular stress including cancer transformation and viral infection. High expression of MICA in cancer tissues or patients' sera is useful for prognostic or follow-up markers in cancer patients. In this study, phage display technology was employed to improve antigen-binding activities of anti-MICA monoclonal antibodies (WW2G8, WW6B7, and WW9B8. The 12 amino acid residues in the complementarity determining regions (CDRs on the V domain of the heavy chain CDR3 (HCDR3 of these anti-MICA antibodies were modified by PCR-random mutagenesis, and phages displaying mutated anti-MICA Fab were constructed. After seven rounds of panning, five clones of phages displaying mutant anti-MICA Fab which exhibited 3–7-folds higher antigen-binding activities were isolated. Two clones of the mutants (phage-displayed mutant Fab WW9B8.1 and phage-displayed mutant Fab WW9B8.21 were confirmed to have antigen-binding specificity for cell surface MICA proteins by flow cytometry. These phage clones are able to recognize MICA in a native form according to positive results obtained by indirect ELISA and flow cytometry. Thus, these phage particles could be potentially used for further development of nanomedicine specifically targeting cancer cells expressing MICA proteins.

  20. Spatio-temporal patterns of Campylobacter colonization in Danish broilers

    DEFF Research Database (Denmark)

    Chowdhury, S; Themudo, G E; Sandberg, M

    2013-01-01

    SUMMARY Despite a number of risk-factor studies in different countries, the epidemiology of Campylobacter colonization in broilers, particularly spatial dependencies, is still not well understood. A series of analyses (visualization and exploratory) were therefore conducted in order to obtain...... a better understanding of the spatial and temporal distribution of Campylobacter in the Danish broiler population. In this study, we observed a non-random temporal occurrence of Campylobacter, with high prevalence during summer and low during winter. Significant spatio-temporal clusters were identified...

  1. Prevalence of Campylobacter concisus in diarrhoea of immunocompromised patients

    DEFF Research Database (Denmark)

    Aabenhus, Rune; Permin, Henrik; On, Stephen L.W.;

    2002-01-01

    The importance of Campylobacter species other than C. jejuni/coli in diarrhoeal disease is largely unknown. We wished to determine the prevalence and clinical presentation of C. concisus infection in patients with enteric disease in a tertiary hospital. Stool specimens were routinely tested...... for the presence of Campylobacter species, by use of the filter isolation method. The medical records of the C. concisus-positive patients were reviewed. Of 224 Campylobacter isolates obtained, 110 were identified as C. concisus. Concomitant infection occurred in only 27% of cases. 13 means of protein profiling we...

  2. Chronic diarrhea associated with Campylobacter jejuni infection in a cat.

    Science.gov (United States)

    Fox, J G; Claps, M; Beaucage, C M

    1986-08-15

    Campylobacter jejuni was isolated from a cat with chronic diarrhea. The diarrheic cat and another cat (which previously had diarrhea) in the same household had bactericidal antibody titers to the C jejuni. Clinical response to antibiotic therapy and not recovering Campylobacter sp from normal feces after treatment also supported the diagnosis of Campylobacter-associated diarrhea. Although the owner had a protracted episode of diarrhea, C jejuni was not isolated from the owner's feces, nor was a bactericidal antibody detected in the owner's serum.

  3. Heat tolerance of dairy lactococcal c2 phages

    DEFF Research Database (Denmark)

    Nielsen, Cecilie Lykke Marvig; Basheer, Aideh; Neve, H.

    2011-01-01

    Nine Lactococcus lactis c2 phages propagated on different hosts were screened for thermal resistance in skimmed milk. Pronounced variations in thermal resistance were found. Three phages displayed high sensitivity towards heat resulting in >8 log reductions after 70 °C for 5 min, whereas the most...

  4. Fluorescent T7 display phages obtained by translational frameshift

    NARCIS (Netherlands)

    Slootweg, E.J.; Keller, H.J.H.G.; Hink, M.A.; Borst, J.W.; Bakker, J.; Schots, A.

    2006-01-01

    Lytic phages form a powerful platform for the display of large cDNA libraries and offer the possibility to screen for interactions with almost any substrate. To visualize these interactions directly by fluorescence microscopy, we constructed fluorescent T7 phages by exploiting the flexibility of pha

  5. Kitchen Practices Used in Handling Broiler Chickens and Survival of Campylobacter spp. on Cutting Surfaces in Kampala, Uganda

    National Research Council Canada - National Science Library

    Irene Wanyenya; Charles Muyanja; George William Nasinyama

    2004-01-01

    .... The survival of Campylobacter spp. on kitchen cutting surfaces was determined by inoculating approximately 106 CFU of Campylobacter jejuni onto sterile plastic, wooden, and metal cutting boards...

  6. A Quantitative Real-Time PCR Approach for Assessing Campylobacter jejuni and Campylobacter coli Colonization in Broiler Herds.

    Science.gov (United States)

    Haas, Katrin; Overesch, Gudrun; Kuhnert, Peter

    2017-04-01

    Human campylobacteriosis is a major public health concern in developed countries, with Campylobacter jejuni and Campylobacter coli from poultry recognized as the main source of human infection. Identification of Campylobacter-positive broiler herds before slaughter is essential for implementing measures to avoid carryover of pathogens via the slaughter process into the food chain. However, appropriate methods that have been validated for testing poultry flocks antemortem are lacking for Campylobacter. A quantitative real-time PCR (qPCR) that allows simultaneous detection and quantification of C. jejuni and C. coli was adapted and optimized to be applied on boot socks. The adjusted qPCR serves as an easy, sensitive, and quantitative method for Campylobacter detection in poultry flocks antemortem by analysis of boot socks. An adequate correlation was found between qPCR and culture, as well as between boot socks and cecal samples, which are regarded as the "gold standard." Therefore, boot sock sampling followed by qPCR analysis provides a reliable and simple method for assessing Campylobacter load within a flock prior to slaughter. The approach allows categorization of broiler herds into negative, low, moderate, or high Campylobacter colonization. Based on the results of this new approach, risk assessment models, such as evaluating the possible effect of sorting flocks before slaughter, can be easily implemented. Similarly, targeted identification of highly colonized flocks for improvement of biosecurity measures at the farm level will become feasible, presenting an opportunity to increase food safety.

  7. Efficacy of a typing scheme for Campylobacter based on the combination of true and questionable CRISPR.

    Science.gov (United States)

    de Cárdenas, Inés; Fernández-Garayzábal, José F; de la Cruz, María-Luisa; Domínguez, Lucas; Ugarte-Ruiz, María; Gómez-Barrero, Susana

    2015-12-01

    This study evaluates an improved scheme for Campylobacter genotyping based on the combination of true and questionable CRISPR (clustered regularly interspaced short palindromic repeats) elements. A total of 180 Campylobacter strains (Campylobacter jejuni n=93 and Campylobacter coli n=87), isolated from neck skin and caecal content of broilers, poultry meat and sewage water were analysed. Another 97 C. jejuni DNA samples from cases of human campylobacteriosis were assessed. Sixty-three genotypes were found in C. jejuni considering only true CRISPR, and 16 additional genotypes were identified when questionable CRISPR were also taken into account. Likewise in C. coli the number of genotypes increased from eight for only true CRISPR to 14 after including questionable CRISPR elements. The number of typeable C. jejuni and C. coli isolates was 115 (60.5%) and 17 (19.5%) respectively considering only true CRISPR. These percentages increased to 92.7% (n=176) and 39.1% (n=34) respectively when both true and questionable CRISPR were considered. 60.9% of the C. coli isolates were non-typeable by CRISPR due to the lack of any PCR amplifiable CRISPR loci, which raises questions about CRISPR analysis as an appropriate method for C. coli typing. However the assessment of true and questionable CRISPR has proved to be fairly useful for typing C. jejuni due to its high discriminatory power (Simpson's index=0.960) and typeability (92.7%) values. The results of the present work show that our genotyping method based on the combination of true and questionable CRISPR elements may be used as a suitable complementary tool to existing C. jejuni genotyping methods. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Phages Preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, Present and Future

    Directory of Open Access Journals (Sweden)

    Annika Gillis

    2014-07-01

    Full Text Available Many bacteriophages (phages have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here.

  9. Prevalence of thermotolerant Campylobacter in broiler flocks and broiler carcasses in Italy

    Directory of Open Access Journals (Sweden)

    Giacomo Migliorati

    2010-12-01

    Full Text Available In accordance with European Union regulations, from 5 February until 15 December 2008, sampling and analysis activities were conducted in Italy to assess the extent of contamination caused by thermotolerant Campylobacter in broiler chickens farmed nationwide. The survey involved 48 poultry slaughterhouses distributed across eleven regions of Italy, where the caeca and carcasses of 393 slaughter batches were sampled. A total of 284 batches (72.3% gave positive results for Campylobacter spp. as follows: 52.1% were contaminated by C. jejuni, 55.6% by C. coli and 1.1% by C. lari. C. jejuni and C. coli were isolated together in 37 batches (13% of positive results. Campylobacter spp. was isolated only from the caeca in 251 slaughter batches (63.9% including caecal isolates of C. jejuni (48.2%, C. coli (50.6%, and C. lari (1.2%. Carcasses from 182 batches (46.3% were contaminated by C. jejuni in 40.7% of cases, C. coli in 57.7% and the absence of C. lari from all batches examined. The contamination level observed in the carcasses ranged between 10 and 1.6 × 107 cfu/g.

  10. Detection of Campylobacter jejuni in rectal swab samples from Rousettus amplexicaudatus in the Philippines.

    Science.gov (United States)

    Hatta, Yuki; Omatsu, Tsutomu; Tsuchiaka, Shinobu; Katayama, Yukie; Taniguchi, Satoshi; Masangkay, Joseph S; Puentespina, Roberto; Eres, Eduardo; Cosico, Edison; Une, Yumi; Yoshikawa, Yasuhiro; Maeda, Ken; Kyuwa, Shigeru; Mizutani, Tetsuya

    2016-09-01

    Bats are the second diversity species of mammals and widely distributed in the world. They are thought to be reservoir and vectors of zoonotic pathogens. However, there is scarce report of the evidence of pathogenic bacteria kept in bats. The precise knowledge of the pathogenic bacteria in bat microbiota is important for zoonosis control. Thus, metagenomic analysis targeting the V3-V4 region of the 16S rRNA of the rectal microbiota in Rousettus amplexicaudatus was performed using high throughput sequencing. The results revealed that 103 genera of bacteria including Camplyobacter were detected. Campylobacter was second predominant genus, and Campylobacter coli and Campylobacter jejuni were identified in microbiome of R. amplexicaudatus. Campylobacteriosis is one of the serious bacterial diarrhea in human, and the most often implicated species as the causative agent of campylobacteriosis is C. jejuni. Therefore, we investigated the prevalence of C. jejuni in 91 wild bats with PCR. As a result of PCR assay targeted on 16S-23S intergenic spacer, partial genome of C. jejuni was detected only in five R. amplexicaudatus. This is the first report that C. jejuni was detected in bat rectal swab samples. C. jejuni is the most common cause of campylobacteriosis in humans, transmitted through water and contact with livestock animals. This result indicated that R. amplexicaudatus may be a carrier of C. jejuni.

  11. Oral Campylobacter species: Initiators of a subgroup of inflammatory bowel disease?

    Science.gov (United States)

    Zhang, Li

    2015-08-21

    In recent years, a number of studies detected a significantly higher prevalence of Campylobacter species such as Campylobacter concisus (C. concisus) in intestinal biopsies and fecal samples collected from patients with inflammatory bowel disease (IBD) compared to controls. Most of these Campylobacter species are not of zoonotic origin but are human oral Campylobacter species. Bacterial species usually cause diseases in the location where they colonize. However, C. concisus and other oral Campylobacter species are associated with IBD occurring at the lower parts of the gastrointestinal tract, suggesting that these Campylobacter species may have unique virulence factors that are expressed in the lower parts of the gastrointestinal tract.

  12. Occurrence of Campylobacter jejuni and Campylobacter coli and their biotypes in beef and dairy cattle from the south of Chile Ocorrência de Campylobacter jejuni e Campylobacter coli e seus biotipos em bovinos de corte e de leite no sul do Chile

    Directory of Open Access Journals (Sweden)

    Heriberto Fernández

    2009-09-01

    Full Text Available The prevalence of Campylobacter jejuni and Campylobacter coli and their biotypes in beef and dairy cattle from the South of Chile was established. Campylobacter were statistically more prevalent among beef cattle (35.9% than among dairy cattle (21.3%, being C. jejuni the species most frequently isolated.Foi estabelecida a prevalência de Campylobacter jejuni e Campylobacter coli e seus biotipos, em bovinos de corte e de leite do sul do Chile. Campylobacter foi estatisticamente mais prevalente nos bovinos de corte (35,9% do que nos bovinos de leite (21,3%, sendo C. jejuni a espécie mais frequentemente isolada.

  13. Antimicrobial resistance profiling and molecular subtyping of Campylobacter spp. from processed turkey

    Directory of Open Access Journals (Sweden)

    Sherwood Julie S

    2009-09-01

    Full Text Available Abstract Background Campylobacter is a major cause of human disease worldwide and poultry are identified as a significant source of this pathogen. Most disease in humans is associated with the consumption of contaminated poultry or cross-contamination with other foods. The primary drugs of choice for treatment of human campylobacteriosis include erythromycin and ciprofloxacin. In this study, we investigated the prevalence of resistance to erythromycin and ciprofloxacin in Campylobacter isolates recovered from turkey carcasses at two processing plants in the Upper Midwest US. Further analysis of a subset of isolates was carried out to assess resistance and genotype profiles. Results Campylobacter isolates from plant A (n = 439; including 196 C. coli and 217 C. jejuni and plant B (n = 362, including 281 C. coli and 62 C. jejuni were tested for susceptibility to ciprofloxacin and erythromycin using agar dilution. C. coli were more frequently resistant than C. jejuni in both plants, including resistance to ciprofloxacin (28% of C. jejuni and 63% of C. coli, plant B; and 11% of C. coli, plant A. Erythromycin resistance was low among C. jejuni (0% plant A and 0.3% plant B compared to C. coli (41%, plant A and 17%, plant B. One hundred resistant and susceptible isolates were selected for additional antimicrobial susceptibility testing, restriction fragment length polymorphism analysis of the flaA gene (fla typing, and pulsed-field gel electrophoresis (PFGE. Fla-PFGE types obtained (n = 37 were associated with a specific plant with the exception of one type that was isolated from both plants. C. coli isolates (n = 65 were grouped into 20 types, while C. jejuni isolates (n = 35 were grouped into 17 types. Most isolates with identical fla-PFGE patterns shared identical or very similar antimicrobial resistance profiles. PFGE alone and composite analysis using fla-PFGE with resistance profiles separated C. jejuni and C. coli into distinct groups. Conclusion

  14. Molecular Typing of Campylobacter jejuni and Campylobacter coli Isolated from Various Retail Meats by MLST and PFGE.

    Science.gov (United States)

    Noormohamed, Aneesa; Fakhr, Mohamed K

    2014-01-08

    Campylobacter species are one of the leading causes of foodborne disease in the United States. Campylobacter jejuni and Campylobacter coli are the two main species of concern to human health and cause approximately 95% of human infections. Molecular typing methods, such as pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) are often used to source track foodborne bacterial pathogens. The aim of the present study was to compare PFGE and MLST in typing strains of C. jejuni and C. coli that were isolated from different Oklahoma retail meat sources. A total of 47 Campylobacter isolates (28 C. jejuni and 19 C. coli) isolated from various retail meat samples (beef, beef livers, pork, chicken, turkey, chicken livers, and chicken gizzards) were subjected to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). PFGE was able to group the 47 Campylobacter isolates into two major clusters (one for C. jejuni and one for C. coli) but failed to differentiate the isolates according to their source. MLST revealed 21 different sequence types (STs) that belonged to eight different clonal complexes. Twelve of the screened Campylobacter isolates (8 C. jejuni and 4 C. coli) did not show any defined STs. All the defined STs of C. coli isolates belonged to ST-828 complex. The majority of C. jejuni isolates belonged to ST-353, ST-607, ST-52, ST-61, and ST-21 complexes. It is worthy to mention that, while the majority of Campylobacter isolates in this study showed STs that are commonly associated with human infections along with other sources, most of the STs from chicken livers were solely reported in human cases. In conclusion, retail meat Campylobacter isolates tested in this study particularly those from chicken livers showed relatedness to STs commonly associated with humans. Molecular typing, particularly MLST, proved to be a helpful tool in suggesting this relatedness to Campylobacter human isolates.

  15. Specific probe selection from landscape phage display library and its application in enzyme-linked immunosorbent assay of free prostate-specific antigen.

    Science.gov (United States)

    Lang, Qiaolin; Wang, Fei; Yin, Long; Liu, Mingjun; Petrenko, Valery A; Liu, Aihua

    2014-03-01

    Probes against targets can be selected from the landscape phage library f8/8, displaying random octapeptides on the pVIII coat protein of the phage fd-tet and demonstrating many excellent features including multivalency, stability, and high structural homogeneity. Prostate-specific antigen (PSA) is usually determined by immunoassay, by which antibodies are frequently used as the specific probes. Herein we found that more advanced probes against free prostate-specific antigen (f-PSA) can be screened from the landscape phage library. Four phage monoclones were selected and identified by the specificity array. One phage clone displaying the fusion peptide ERNSVSPS showed good specificity and affinity to f-PSA and was used as a PSA capture probe in a sandwich enzyme-linked immunosorbent assay (ELISA) array. An anti-human PSA monoclonal antibody (anti-PSA mAb) was used to recognize the captured antigen, followed by horseradish peroxidase-conjugated antibody (HRP-IgG) and o-phenylenediamine, which were successively added to develop plate color. The ELISA conditions such as effect of blocking agent, coating buffer pH, phage concentration, antigen incubation time, and anti-PSA mAb dilution for phage ELISA were optimized. On the basis of the optimal phage ELISA conditions, the absorbance taken at 492 nm on a microplate reader was linear with f-PSA concentration within 0.825-165 ng/mL with a low limit of detection of 0.16 ng/mL. Thus, the landscape phage is an attractive biomolecular probe in bioanalysis.

  16. Genomic and Functional Analyses of Rhodococcus equi Phages ReqiPepy6, ReqiPoco6, ReqiPine5, and ReqiDocB7 ▿

    Science.gov (United States)

    Summer, E. J.; Liu, M.; Gill, J. J.; Grant, M.; Chan-Cortes, T. N.; Ferguson, L.; Janes, C.; Lange, K.; Bertoli, M.; Moore, C.; Orchard, R. C.; Cohen, N. D.; Young, R.

    2011-01-01

    The isolation and results of genomic and functional analyses of Rhodococcus equi phages ReqiPepy6, ReqiDocB7, ReqiPine5, and ReqiPoco6 (hereafter referred to as Pepy6, DocB7, Pine5, and Poco6, respectively) are reported. Two phages, Pepy6 and Poco6, more than 75% identical, exhibited genome organization and protein sequence likeness to Lactococcus lactis phage 1706 and clostridial prophage elements. An unusually high fraction, 27%, of Pepy6 and Poco6 proteins were predicted to possess at least one transmembrane domain, a value much higher than the average of 8.5% transmembrane domain-containing proteins determined from a data set of 36,324 phage protein entries. Genome organization and protein sequence comparisons place phage Pine5 as the first nonmycobacteriophage member of the large Rosebush cluster. DocB7, which had the broadest host range among the four isolates, was not closely related to any phage or prophage in the database, and only 23 of 105 predicted encoded proteins could be assigned a functional annotation. Because of the relationship of Rhodococcus to Mycobacterium, it was anticipated that these phages should exhibit some of the features characteristic of mycobacteriophages. Traits that were identified as shared by the Rhodococcus phages and mycobacteriophages include the prevalent long-tailed morphology and the presence of genes encoding LysB-like mycolate-hydrolyzing lysis proteins. Application of DocB7 lysates to soils amended with a host strain of R. equi reduced recoverable bacterial CFU, suggesting that phage may be useful in limiting R. equi load in the environment while foals are susceptible to infection. PMID:21097585

  17. Genomic sequence and activity of KS10, a transposable phage of the Burkholderia cepacia complex

    Directory of Open Access Journals (Sweden)

    Shrivastava Savita

    2008-12-01

    Full Text Available Abstract Background The Burkholderia cepacia complex (BCC is a versatile group of Gram negative organisms that can be found throughout the environment in sources such as soil, water, and plants. While BCC bacteria can be involved in beneficial interactions with plants, they are also considered opportunistic pathogens, specifically in patients with cystic fibrosis and chronic granulomatous disease. These organisms also exhibit resistance to many antibiotics, making conventional treatment often unsuccessful. KS10 was isolated as a prophage of B. cenocepacia K56-2, a clinically relevant strain of the BCC. Our objective was to sequence the genome of this phage and also determine if this prophage encoded any virulence determinants. Results KS10 is a 37,635 base pairs (bp transposable phage of the opportunistic pathogen Burkholderia cenocepacia. Genome sequence analysis and annotation of this phage reveals that KS10 shows the closest sequence homology to Mu and BcepMu. KS10 was found to be a prophage in three different strains of B. cenocepacia, including strains K56-2, J2315, and C5424, and seven tested clinical isolates of B. cenocepacia, but no other BCC species. A survey of 23 strains and 20 clinical isolates of the BCC revealed that KS10 is able to form plaques on lawns of B. ambifaria LMG 19467, B. cenocepacia PC184, and B. stabilis LMG 18870. Conclusion KS10 is a novel phage with a genomic organization that differs from most phages in that its capsid genes are not aligned into one module but rather separated by approximately 11 kb, giving evidence of one or more prior genetic rearrangements. There were no potential virulence factors identified in KS10, though many hypothetical proteins were identified with no known function.

  18. γ-PGA Hydrolases of Phage Origin in Bacillus subtilis and Other Microbial Genomes.

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    Stefania Mamberti

    Full Text Available Poly-γ-glutamate (γ-PGA is an industrially interesting polymer secreted mainly by members of the class Bacilli which forms a shield able to protect bacteria from phagocytosis and phages. Few enzymes are known to degrade γ-PGA; among them is a phage-encoded γ-PGA hydrolase, PghP. The supposed role of PghP in phages is to ensure access to the surface of bacterial cells by dismantling the γ-PGA barrier. We identified four unannotated B. subtilis genes through similarity of their encoded products to PghP; in fact these genes reside in prophage elements of B. subtilis genome. The recombinant products of two of them demonstrate efficient polymer degradation, confirming that sequence similarity reflects functional homology. Genes encoding similar γ-PGA hydrolases were identified in phages specific for the order Bacillales and in numerous microbial genomes, not only belonging to that order. The distribution of the γ-PGA biosynthesis operon was also investigated with a bioinformatics approach; it was found that the list of organisms endowed with γ-PGA biosynthetic functions is larger than expected and includes several pathogenic species. Moreover in non-Bacillales bacteria the predicted γ-PGA hydrolase genes are preferentially found in species that do not have the genetic asset for polymer production. Our findings suggest that γ-PGA hydrolase genes might have spread across microbial genomes via horizontal exchanges rather than via phage infection. We hypothesize that, in natural habitats rich in γ-PGA supplied by producer organisms, the availability of hydrolases that release glutamate oligomers from γ-PGA might be a beneficial trait under positive selection.

  19. Filamentous Phages As a Model System in Soft Matter Physics.

    Science.gov (United States)

    Dogic, Zvonimir

    2016-01-01

    Filamentous phages have unique physical properties, such as uniform particle lengths, that are not found in other model systems of rod-like colloidal particles. Consequently, suspensions of such phages provided powerful model systems that have advanced our understanding of soft matter physics in general and liquid crystals in particular. We described some of these advances. In particular we briefly summarize how suspensions of filamentous phages have provided valuable insight into the field of colloidal liquid crystals. We also describe recent experiments on filamentous phages that have elucidated a robust pathway for assembly of 2D membrane-like materials. Finally, we outline unique structural properties of filamentous phages that have so far remained largely unexplored yet have the potential to further advance soft matter physics and material science.

  20. Chemical posttranslational modification of phage-displayed peptides.

    Science.gov (United States)

    Ng, Simon; Tjhung, Katrina F; Paschal, Beth M; Noren, Christopher J; Derda, Ratmir

    2015-01-01

    Phage-displayed peptide library has fueled the discovery of novel ligands for diverse targets. A new type of phage libraries that displays not only linear and disulfide-constrained cyclic peptides but moieties that cannot be encoded genetically or incorporated easily by bacterial genetic machinery has emerged recently. Chemical posttranslational modification of phage library is one of the simplest approaches to encode nonnatural moieties. It confers the library with new functionality and makes it possible to select and evolve molecules with properties not found in the peptides, for instance, glycopeptides recognized by carbohydrate-binding protein and peptides with photoswitching capability. To this end, we describe the newly emerging techniques to chemically modify the phage library and quantify the efficiency of the reaction with a biotin-capture assay. Finally, we provide the methods to construct N-terminal Ser peptide library that allows site-selective modification of phage.