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Coinfections

2012

German National Academy of Sciences Leopoldina, Halle (Saale), Germany

June 7th to 8th 2012 www.coinfections2012.com

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About the European Academy of Microbiology (EAM) One of the goals of FEMS has been to amplify the impact of microbiology and microbiologists in Europe. For this purpose, it is important to establish a leadership group of European microbiologists that will be in close association with FEMS. As a result, FEMS executives established an “Academy” of senior microbiologists within Europe with the objective of providing a source of

advice for the Federation and a united advisory source for governmental and other bodies. The European Academy of Microbiology promotes excellence in microbiology in Europe through targeted programs and activities. Members of the academy are elected through a highly selective, peer-reviewed process, based on their records of scientific achievement and original contributions that have advanced microbiology.

EAM Board Members

Philippe Sansonetti (France) EAM president

Hans Dieter Klenk (Germany) EAM board member

Tone Tønjum (Norway) EAM board member

Eliora Ron (Israel) EAM Secretary

Bernhard Schink (Germany) FEMS president

Hillary Lappin-Scott (UK) EAM board member

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Scope This scientific event highlights recent advances in our understanding of polymicrobial infections, synergies between microbes and relevant aspects of symbiosis. Scientists at the international forefront will give presentations and contribute to the discussions. Also, there will be posters presented by meeting participants. The presentations will cover the entire field from molecules to medicine. Simultaneous infection of a host by multiple and synergistic pathogens, incremental infections arising by initial infection followed by superinfection and simultaneous coinfection of a single cell occur where one otherwise minor organism unleashes the full pathogenic potential of an established pathogen. When pathogens enforce each other synergistically, disease transmission and progression are enhanced, in syndemism. One prime example of such a globally common coinfection to be highlighted at the meeting involves both tuberculosis and HIV, where one is exacerbating the other in the pandemic. Coinfections arise with representatives of all classes of microbes, bacteria, viruses, yeast/ fungi and parasites as well as combinations of these. Coinfections can also affect each other negatively in microbial interference where one bacterial species suppresses the virulence or colonization of the bacteria, alleviating the pathogenic pressure. One example of the latter is when Pseudomonas aeruginos suppresses pathogenic Staphylococcus aureus colony formation. The global prevalence of coinfections is commonplace, but the impact of coinfections is greatly underestimated and the actual numbers are currently non-available. We will therefore also elucidate some of the societal impact of coinfections and disease and discuss controversies in regard to pathogenesis, diagnostics and treatment. The multifaceted approach to the emerging field of coinfections makes this conference a “must” for students and researchers in the infection biology sciences.

Scientific committee: Hans Dieter Klenk, Eliora Ron, Philippe Sansonetti and Tone Tønjum

Organizing committee: Barbara Dartee, Anne Haukvik and Chared Verschuur

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PROGRAM | FEMS/Leopoldina - Halle (Saale), Germany European Academy of Microbiology (EAM) meeting

Thursday June 7th 13:00-14:00 Lunch 14:00-14:10 Welcome to Coinfections2012: Philippe Sansonetti/Hans Dieter Klenk Session 1: Microbial communities I: Complex microbial communities and gene and signal exchanges Chair: Tone Tønjum 14:10-14.40 Albert Osterhaus, Erasmus Medical Center, Rotterdam The Netherlands: Emergin virus infections in a changing world 14:40-15:10

Søren Molin, The Technical University of Denmark, Lyngby, Denmark: Adaptive evolution of Pseudomonas aeruginosa populations in airways of cystic fibrosis patients

Session 2: Bacterial/viral symbionts of bacteria - Co-infections with parasites Chair: hans Dieter Klenk 15:10-15:40

Nicolas Fasel, University of Lausanne, Switzerland: A tale of a virus and of a parasite

15:40- 16:10

Achim Hoerauf, University Bonn Medical Center Munich, Germany: Filarial nematodes and their Wolbachia endosymbionts unleash the immune response eliciting clinical filariasis

16:10- 16:40

Coffee break

Session 3: Significance of co-infections – Epidemiology Chair: Philippe Sansonetti 16:40-17:10

Birgitta Henriques Normark, Karolinska Institute, Stockholm, Sweden: Pneumococcal-influenza co-infections

17:10-17:20 Brian Davis, University of Michigan, US: Influenza and Community-acquired Pneumonia Interactions: The Impact of Order and Time of Infection on Population Patterns 17:20-17:30

Eva Böttcher-Friebertshäuser, Institut für Virologie, Philipps-Universität Marburg, Germany: Activation of influenza viruses by proteases from host cells and bacteria

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Session 4: Viral modulation of the immune response to bacterial infections 17:30-17:50

Robert J. Wilkinson, University of Cape Town, South Africa, Imperial College London and MRC National Institute for Medical Research, UK: HIV-1 and the immune response to tuberculosis ----

17:50-18:10 Patrik M. Bavoil, University of Maryland School of Dentistry, US: Launching Pathogens & Disease (PAD), a new journal of FEMS 18:15-

Poster session

Dinner

Friday June 8th Session 5: Microbial interactions Chair: Hans Dieter Klenk 09:00-09:30 Jonathan McCullers, University of Tennessee Health Sciences Center and St. Jude Children’s Research Hospital, Memphis, US: From viruses to worms: How pathogen-associated innate immune defects facilitate bacterial pneumonia 09:30-10:00 Linda Wammes, Leiden University Medical Center, Leiden, The Netherlands: Immunological consequences of helminth-malaria co-infections 10:00-10:30

Coffee break & posters

Session 6: Microbial communities II: Microbiota in infections & Susceptibility to infections 10:45-11:30 Julie Pfeiffer, University of Texas Southwestern Medical Centre, Dallas, US: Intestinal microbiota promote enteric virus infections 11:30-12:00

Patrik M. Bavoil, University of Maryland School of Dentistry, US: Eco-Pathogenomics of Chlamydial Reproductive Tract Infection

Concluding remarks: Philippe Sansonetti/Hans Dieter Klenk 12:00-13:00 Lunch

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About the speakers

Albert Osterhaus, Professor, Head of the Institute of Virology of the Erasmus Medical Center, Rotterdam, The Netherlands

Søren Molin, Professor, The Technical University of Denmark, Lyngby, Denmark

EMERGING VIRUS INFECTIONS IN A CHANGING WORLD

ADAPTIVE EVOLUTION OF PSEUDOMONAS AERUGINOSA POPULATIONS IN AIRWAYS OF CYSTIC FIBROSIS PATIENTS

Dr. Albert Osterhaus is one of the world’s leading virologists and his group was the first to identify human infection with the avian influenza strain H5N1. His first major breakthrough came in 1998 when he identified a new morbilivirus that caused a mass dieoff of seals in Northwestern Europe. In 1997, his group discovered that a Hong Kong flu strain that had killed a three-year-old boy belonged to an avian influenza strain called H5N1. He was also the first scientist to show that H5N1 can be transferred into humans. In 2000 he and his team identified Influenza B virus, a type of virus that normally infects only humans in seals off the coast of the Netherlands. In 2001, his group identified human metapneumovirus (hMPV), which causes a spectrum of respiratory illnesses ranging from mild upper respiratory tract infections to severe bronchiolitis and pneumonia. In 2003, at the height of the panic over SARS (Severe Acquired Respiratory Syndrome) in Hong Kong, he again showed his skill at moving fast to tackle a serious problem. Within three weeks he had proved that the disease was caused by a newly discovered coronavirus that resides in civet cats, other carnivorous animals or bats. Currently he heads a 100-strong lab at Erasmus MC, Rotterdam, is the co-founder of two biotech companies, and is part of numerous global collaborations. He is particularly interested in viruses that cross species barriers, are highly pathogenic and which cause disease globally viruses such as HIV, SARS CoV and influenza viruses.

Patients suffering from the genetic disorder cystic fibrosis (CF) aquire airway infections at a young age, and several microorganisms are usually identified in the patients. Peudomonas aeruginosa is assumed to be the major cause of morbidity and mortality, but it cannot be excluded that other infecting microorganisms play a significant role in the progression of the lung disease. Micro-evolution studies have shown that during the life span of most CF patients there is a continuous adaptation of specific lineages of P. aeruginosa, which colonize the patient airways in early childhood and stay associated with the patients for the rest of their lives. This adaptation is normally interpreted to reflect the specific conditions of this particular host/ microbe relationship, but it is possible that also microbe/ microbe interactions impact on the evolutionary routes. Genomic and functional studies of several isolates have provided insight into the evolutionary trajectories by which the bacteria gradually adapt to the environmental conditions in the CF airways. From these and other data we have derived an evolutionary ’road-map’ explaining how P. aeruginosa in different ways can attain life-styles compatible with the stressful airway environment of CF patients continuously treated with antibiotics and harbouring a complex micro-flora of both aerobic and anaerobic bacteria and fungi. In addition to an increased understanding of biological evolutionary mechanisms we think that our results and conclusions may also pave the way for improved treatment of the CF airway infections.

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Nicolas Fasel, Professor, University of Lausanne, Switzerland

Achim Hoerauf, Professor, University Bonn Medical Center Munich, Germany

A TALE OF A VIRUS AND OF A PARASITE

FILARIAL NEMATODES AND THEIR WOLBACHIA ENDOSYMBIONTS UNLEASH THE IMMUNE RESPONSE ELICITING CLINICAL FILARIASIS

In Latin America, Leishmania Viannia (L.Viannia) subgenus species cause cutaneous (CL) and mucocutaneous (MCL) leishmaniasis. Approximately, 5-10% of individuals asymptomatic or with resolved CL lesions can develop MCL, which is associated with persistent immune responses showing pro-inflammatory mediators and dissemination of the infection. Hamsters infected with L.Viannia parasites isolated from human MCL lesions reproduce the metastatic phenotype with primary and secondary lesion development. Using this model, clones derived from metastasizing L. guyanensis strains were characterised as being metastatic (L.g.M+), or nonmetastatic (L.g.M-) after infection, depending on their ability to reproducibly develop secondary metastatic lesions. We showed that metastasizing parasites have a high Leishmania RNA Virus-1 (LRV1) burden that is recognized by the host Toll-Like receptor 3 (TLR3) to induce pro-inflammatory cytokines and chemokines. Paradoxically, these TLR3-mediated immune responses rendered mice more susceptible to infection and the animals developed an increased footpad swelling and parasitemia. We are currently evaluating the prevalence of LRV in different human isolates, the innate and acquired immune response to LRV as well as understanding the immunological mediators implicated in the dissemination of the infection. Our results open the doorway to better diagnosis of risk for MCL disease and facilitate the development of new, and more efficient treatment regimes.

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About the speakers

Birgitta Henriques Normark, Professor, Karolinska Institute, Stockholm, Sweden

Robert J. Wilkinson, Professor, University of Cape Town, South Africa, Imperial College London and MRC National Institute for Medical Research, UK

PNEUMOCOCCAL-INFLUENZA CO-INFECTIONS

HIV-1 AND THE IMMUNE RESPONSE TO TUBERCULOSIS

Streptococcus pneumoniae is a major contributor to morbidity and mortality worldwide, causing 1.5-2 million deaths yearly, which is within the same range as mortality associated with tuberculosis. Pneumococci are the most common cause of otitis media, sinusitis and community acquired pneumonia, but also a major cause of septicemia and meningitis. Despite being a devastating pathogen, pneumococci are also found to colonize healthy children and up to 60% of children attending daycare centers may harbor these bacteria in the nasopharynx without disease. The major risk groups for developing a pneumococcal invasive disease are small children and the elderly, as well as immunocompromised individuals. Prior virus infections, especially influenza A virus (IAV) infections, also sensitize the host. The mechanisms for this sensitization are not fully understood. Influenza viruses cause common infections that are spread around the world in seasonal epidemics, resulting in the deaths of between 250,000 and 500,000 people every year and up to millions in some pandemic years. Infection with IAV alone is rarely fatal. Instead, death associated with IAV is usually mediated by super-infection with bacteria, mainly S. pneumoniae. Data on the epidemiology as well as novel mechanistic data of pneumococcal-influenza coinfections will be provided.

HIV associated tuberculosis is one of the greatest health challenges on the continent of Africa. Over the last 7 years we have contributed to studies of interferon gamma release assays in HIV infected patients. We have investigated the effects and mechanisms of preventive therapies (antiretroviral therapy, isoniazid and vitamin D). We have determined that protective antiretroviral mediated immune recovery in HIV-TB is associated with expansion of central memory T cells rather than the commonly determined effector response. Conversely pathological immune restoration, as exemplified by the HIV-TB immune reconstitution inflammatory syndrome, is contributed to by dysregulated Th1 expansions and by exaggerated cytokine release. We have determined by clinical trials that corticosteroids help suppress pathological immunity via downregulation of IL-6 and TNF. We have recently documented that severe vitamin D deficiency is highly prevalent amongst black Africans and associated with HIV-TB. We have recently completed a clinical trial in which 1539 HIV infected persons were randomised to antiretrovirals vs antiretrovirals plus isoniazid for the prevention of TB. We have also enrolled 104 of a projected 940 HIV infected persons to an ongoing phase IIB trial of a novel tuberculosis vaccine.

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Patrik M. Bavoil, University of Maryland School of Dentistry, US

Jonathan McCullers, Professor and Chair of Pediatrics University of Tennessee Health Sciences Center and St. Jude Children’s Research Hospital, US

LAUNCHING PATHOGENS & DISEASE (PAD), A NEW JOURNAL OF FEMS

FROM VIRUSES TO WORMS: HOW PATHOGENASSOCIATED INNATE IMMUNE DEFECTS FACILITATE BACTERIAL PNEUMONIA

ECO-PATHOGENOMICS OF CHLAMYDIAL REPRODUCTIVE TRACT INFECTION The Bavoil laboratory studies the pathogenesis of the obligate intracellular pathogen, Chlamydia, and its bacteriophages. Specific research areas include the role of Chlamydia type III secretion in pathogenesis and development, the impact of Chlamydia phage infection on disease, the role of the polymorphic membrane protein family of C. trachomatis in infection and disease and comparative genomics within the Chlamydiaceae. Drs. Patrik Bavoil and Jacques Ravel are co-Directors of the NIH-NIAID-funded STI Cooperative Research Center “Ecopathogenomics of Chlamydial Reproductive Tract Infection” (EPCRTI), also known as the CHARM study, and of the contract “A Systems Biology Approach to Chlamydia trachomatis Persistent/Recurrent Infection”.

Bacterial pneumonia is the leading cause of death in children under 5 years of age. Fatal infections are typically seen in hosts compromised by either chronic medical illnesses or co-infections. The mechanisms by which coinfecting pathogens alter innate immune responses to bacteria is an area of great interest and of active study. In this seminar, Dr. McCullers will review recent progress in his laboratory towards understanding immune defects engendered by two disparate co-infections, influenza viruses and helminths. Through divergent pathways, both can alter innate immune responses to Streptococcus pneumoniae and facilitate fatal bacterial pneumonia. The implications of these findings for prevention and treatment of lung disease will be discussed.

Dr. Bavoil is Chief-Editor of the journal FEMS Immunology and Medical Microbiology, soon to be relaunched as “Pathogens and Disease” (01/01/13) and President of the Chlamydia Basic Research Society (CBRS).

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About the speakers

Linda Wammes, MSc MD, Leiden University Medical Center, Leiden, The Netherlands

Julie Pfeiffer, Assistant professor, University of Texas Southwestern Medical Centre, Dallas, US

IMMUNOLOGICAL CONSEQUENCES MALARIA CO-INFECTIONS

HELMINTH-

INTESTINAL MICROBIOTA PROMOTE ENTERIC VIRUS INFECTIONS

Helminths affect more than one billion people worldwide and predominantly reside in (sub-)tropical areas. Although helminths generally do not kill people, they can cause mild to severe morbidities, due to their persistent stay in the human body for years. Helminths can strongly influence the host immune system by driving polarized type 2 responses, accompanied by high levels of circulating IgE and eosinophilia. In addition, helminths are regarded as masters of regulation, as they manipulate the immune system to prevent strong immunity to ensure their own survival. This results in a general T cell hyporesponsiveness that not only affects responses to helminth antigens, but also bystander antigens, such as allergens, vaccines and other pathogens. It is hypothesized that helminths can do so by the generation of a regulatory network, existing of different regulatory subsets, i.e. regulatory T and B cells and alternatively activated macrophages. Part of the research in our group is focusing on how helminth-induced immune modulation affects immune responses and clinical symptoms of coinfections such as malaria, a common phenomenon in tropical areas. Insight in these processes may help to develop strategies for vaccination against malaria and mass deworming programs.

Enteric viruses encounter a vast microbial community in the mammalian digestive tract. However, the effect of the intestinal microbiota on enteric viruses is not well understood. Using mouse models, it was recently shown that intestinal bacteria promote infection with three unrelated enteric viruses, poliovirus, reovirus, and mouse mammary tumor virus. The enhanced replication and pathogenesis of these viruses in microbe-containing mice could occur through microbe-dependent effects on the host and/or virus. For poliovirus, data suggest that bacterial products directly interact with virus particles and increase viral infectivity. Specifically, bacterial surface polysaccharides, such as lipopolysaccharide and peptidoglyan, enhance poliovirus infectivity by enhancing virion stability and aiding attachment to host cells. Overall, enteric viruses may have evolved mechanisms to use gut microbes as an environmental sensor to initiate replication at the optimal site in the intestine.

OF

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Oral poster presentations

Eva Böttcher-Friebertshäuser, Institut für Virologie, Philipps-Universität Marburg, Germany ACTIVATION OF INFLUENZA VIRUSES BY CELLULAR AND BACTERIAL PROTEASES Eva Böttcher-Friebertshäuser, Hans-Dieter Klenk and Wolfgang Garten. Institute of Virology, PhilippsUniversity Marburg, Germany Influenza viruses cause acute infection of the respiratory tract that affects millions of people during seasonal outbreaks and occasional pandemics. Bacterial coinfections contribute significantly to severe pneumonia and deaths during influenza epidemics and pandemics and most commonly isolated bacteria are Staphylococcus aureus, Streptococcus pneumoniae or Haemophilus influenzae. Viral infection is initiated by the surface glycoprotein hemagglutinin (HA) that mediates binding to cell surface receptors and fusion of the virus envelope with the endosomal membrane in order to release the viral genome into the target cell. HA is synthesized as a precursor protein HA0 and has to be cleaved by an exogenous protease into the subunits HA1 and HA2 to gain its fusion capacity. Most influenza viruses require activation by trypsin-like proteases and we identified the transmembrane proteases TMPRSS2 (transmembrane protease, serine S1 member 2) and HAT (human airway trypsin-like protease) as relevant enzymes in the human airway epithelium. TMPRSS2 cleaves HA within the cell during its transport to the plasma membrane, whereas HAT is expressed as an enzymatically active protease on the cell surface that mediates activation of HA during assembly and budding as well as prior to entry into new cells. Interestingly, a protease secreted by S. aureus has been shown to cleave HA in vitro and to increase pathogenicity upon co-infection in mice. Furthermore, streptokinase and staphylokinase support activation of HA via conversion of plasminogen to plasmin, which cleaves the HA of some influenza strains.

Brian Davis, University of Michigan, US

INFLUENZA AND COMMUNITY-ACQUIRED PNEUMONIA INTERACTIONS: THE IMPACT OF ORDER AND TIME OF INFECTION ON POPULATION PATTERNS Brian M. Davis, Allison E. Aiello, Suzanne Dawid, Pejman Rohani, Sourya Shrestha, and Betsy Foxman Discoveries made during the 1918 influenza A pandemic and reports of severe disease associated with coinfection during the 2009 hemagglutinin type 1 and neuraminidase type 1 (commonly known as H1N1 or swine flu) pandemic have renewed interest in the role of coinfection in disease pathogenesis. The authors assessed how various timings of coinfection with influenza virus and pneumoniacausing bacteria could affect the severity of illness at multiple levels of interaction, including the biologic and population levels. Animal studies most strongly support a single pathway of coinfection with influenza inoculation occurring approximately 7 days before inoculation with Streptococcus pneumoniae, but less-examined pathways of infection also may be important for human disease. The authors discussed the implications of each pathway for disease prevention and what they would expect to see at the population level if there were sufficient data available. Lastly, the authors identified crucial gaps in the study of timing of coinfection and proposed related research questions. Am J Epidemiol. 2012 Mar 1;175(5):363-7. Epub 2012 Jan 12.

In conclusion, both cellular and bacterial proteases may contribute to the propagation and pathogenicity of coinfections of influenza virus and certain bacteria.

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Poster abstracts

MULTIPLE INFECTIONS SPECIES IN CATS

WITH MORE

PARVOVIRUS

Mara Battilani, Andrea Balboni, Massimo Giunti. Department of Veterinary Medical Sciences, Alma Mater Studiorum, University of Bologna, Italy. Parvoviruses of carnivores include three closely related autonomous parvoviruses: canine parvovirus (CPV), feline panleukopenia virus (FPV) and mink enteritis virus (MEV). These viruses cause a variety of serious diseases, especially in young patients, since they have a remarkable predilection for replication in rapidly dividing cells. FPV is not the only parvovirus species which infects cats; in addition to MEV, the new variants of canine parvovirus, CPV-2a, 2b and 2c have also penetrated the feline host-range, and they are able to infect and replicate in cats, causing diseases indistinguishable from feline panleukopenia. Fmthennore, as cats are susceptible to both CPV-2 and FPV viruses, superinfection and coinfection with multiple parvovirus strains may occur, potentially facilitating recombination and high genetic heterogeneity. In this study we reported the first clinical case of co-infection with canine parvovirus (CPV) type 2a and feline panleukopenia virus (FPV) with the presence of several viral variants in a not vaccinated 3-months­ yrs-old male kitten with signs of gastroenteritis. Clinical details and haematological findings were supplied in addition to virological results already published, with the aim to investigate the outcome ofparvovirus multiple infections. CPV represent a novel pathogen for cats and may have a synergistic action with FPV, it could alter the clinical course ofthe classical feline panleukopenia, with the acceleration and increase of the seriousness of the infection. The presence of two parvovirus species in the same host increase the chance to establishes persistent infection and stresses the centrality of cat in the epidemiology ofparvovirus of carnivores as source of genetic complexity and emergence ofnew variants. In conclusion, this study provides new important results about the evolutionary dynamics of CPV infections in cats, showing that CPV has presumably started a new process of readaptation in feline hosts.

COINFECTION WITH DIFFERENT CORONAVIRUS IN R. ferrumequinum IN ITALY 1 2 Andrea Balboni ,Jan Felix Drexler , Mara Battilani 1 • 1

Department of Veterinary Medical Sciences, Alma Mater Studiorum, University of Bologna, Italy.

2

Institute of Virology, University of Bonn Medical Centre, Germany.

Coronaviruses (CoVs) are subdivided into four genera. All the mammalian CoVs belong to the first two genera, Alpha­ and Betacoronavirus , whereas all avian CoVs belong to the Gamma- and Deltacoronavirus. In the last few years it was discovered a worldwide association between Alpha- and Betacoronavirus and bats which has led to suppose that all mammalian coronaviruses derive to ancestral viruses residing in bats. Fmthermore, it has also been shown that bats might be the natural reservoir for several viruses closely related genetically to the aetiologic agent of the severe acute respiratory syndrome (SARS..CoV), known as SARS-Iike coronaviruses (SARS-like CoVs) and belonging to the genus Betacoronavirus. Recently, a SYBR Green real-time PCR assay specific for SARS-related CoVs was applied to 45 bats (Rhinolophusferrumequinum) faecal samples collected in Italy in 2009 and resulted in a prevalence of 42 %. The successive sequencing of a fragment of RNAdependent RNA polymerase gene (RdRp), frequently used for phylogenetic analysis, has allowed us to identify the presence of two coronaviruses belonging to different genus, Alpha­ and Betacoronavirus respectively, in the same bat faecal sample. Bioinformatic analysis showed a strict correlation of the betacoronavirus with SARSlike CoVs and a high identity of the alphacoronavirus with other European, African and Asian strains. Coinfections by different coronaviruses are the basis of the recombination phenomena that contribute to the evolution of these viruses. In pmticular, the presence in the same bat of a SARS-like CoV with other coronaviruses stresses the possibility that recombinant events may have a role in the evolutionary process of SARS-related coronaviruses.

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INFLUENZA AND COMMUNITY-ACQUIRED PNEUMONIA INTERACTIONS: THE IMPACT OF ORDER AND TIME ON POPULATION PATTERNS Brian M. Davis, Allison E. Aiello, Suzanne Dawid, Pejman Rohani, Sourya Shrestha, and Betsy Foxman School of public Health, University of Michigan, US Discoveries made during the 1918 influenza A pandemic and reports of severe disease associated with coinfection during the 2009 hemagglutinin type 1 and neuraminidase type 1 (commonly known as H1N1 or swine flu) pandemic have renewed interest in the role of coinfection in disease pathogenesis. The authors assessed how various timings of coinfection with influenza virus and pneumoniacausing bacteria could affect the severity of illness at multiple levels of interaction, including the biologic and population levels. Animal studies most strongly support a single pathway of coinfection with influenza inoculation occurring approximately 7 days before inoculation with Streptococcus pneumoniae, but less-examined pathways of infection also may be important for human disease. The authors discussed the implications of each pathway for disease prevention and what they would expect to see at the population level if there were sufficient data available. Lastly, the authors identified crucial gaps in the study of timing of coinfection and proposed related research questions. Am J Epidemiol. 2012 Mar 1;175(5):363-7. Epub 2012 Jan 12.

COHABITATION IN ENVIRONMENT.

THE

CYSTIC

FIBROSIS

LUNG

Fothergill JL1, Liu X2, Addison RJ2, Walshaw MJ3, Ledson MJ3, Southern K4, McNamara P4, Brockhurst MA2, Winstanley C1. 1.Institute of Infection and Global Health, University of Liverpool, UK 2. Institute of Integrative Biology, University of Liverpool, UK. 3. Amanda Unit, Liverpool Heart and Chest Hospital, UK. 4. Alder Hey Children’s Hospital, Liverpool, UK. Bacteria in the natural environment form complex, interacting communities. This is also true for bacterial infections of animals and humans, where much of the diversity that we see may be driven by microbes interacting with each other as well as with the host. The Cystic Fibrosis (CF) lung facilitates the cohabitation of diverse microbial organisms and we are only just beginning to understand the extent of this diversity. In order to characterise the CF lung microbiome, matched sputum and bronchoalveolar lavage (BAL) samples from three children were sequenced using 454 pyrosequencing of 16S rRNA sequences. This enabled the microbial community in each sample to be identified and compared to determine how representative sputum samples are of the lower lung. Furthermore, BAL samples from the left and right lung were also compared to determine spatial differences in the CF lung microbial population. Considerable differences in the microbial population could be found between samples. A retrospective analysis of published data was performed to further describe the CF lung meta-community. This revealed an extremely diverse range of bacteria that are capable colonising the CF lung, with 317 species from 172 different genera identified. These data will be used to develop a multispecies biofilm model in which microbial interactions (both antagonistic and synergistic) can be investigated. Understanding these complex interactions may uncover novel therapeutic targets and ultimately lead to altered CF patient management.

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Poster abstracts

SYNTHESIS IOF A FLUORESCENT CHITOSAN DERIVATIVE AND ITS APPLICATION AS ANTIBACTERIAL AGENT

PLANT EXTRACTS APPLICATION FOR SALMONELLA CONTROL IN EGG FARMS

2 Shaaban H. Moussa1 ; Klaus Opwis ; Asmaa S. Farouk3

2 1 Ahmed A. Tayel ; Wael F. El-Tras ; Shaaban H. Moussa1

1Genetic Engineering and Biotechnology Research Institute, Minoufiya University, El-Sadat City, Egypt.

1

2DTNW- Deutsches Textilforschungszentrum NordWest e. V Institut an der, Universitat Duisburg 足 Essen, Krefeld, Germany

Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt

3National Research Center, Textile Research Division, Dokki, Cairo, Egypt Chitosan cationic polysaccharide has been reported to be a non-toxic biodegradable antibacterial agent which supports its usage in fields of biotechnology, medications. cosmetology, pharmacology and Fluorescein isothiocyanate-labeled chitosan (FITC-CTS) was synthesized as a macromolecular fluorophore to investigate the molecular mechanism of interaction between chitosan and bacterial cell wall. As a preliminary study, we investigated the antibacterial performance of (FITC-CTS) against Escherichia coli and Micrococcus lutues. Fluorescence imaging was used to obtain high- resolution images of the effect of chitosan on the bacterial morphology. The fluorescence observations of the effect of chitosan as antibacterial agent were correlated with viable cell number.

ENTERIC

Genetic Engineering and Biotechnology Research Institute, Minufiya University, P. 0. box 79/22857, El-Sadat City, Egypt

2

Human illness caused by infection with enteric Salmonella dramatically increased worldwide since their first epidemiological tracking in the mid1970s. Salmonella enterica serovar Typhimurium (S. Typhimurium) and serovar Enteritidis (S. Enteritidis) are the primary and predominant causes of foodborne salmonellosis, that routinely contaminate eggs, in the world. The hen house environment and the contaminated eggshells are from the main routes for Salmonella infection and transmission. The discovery and application of natural alternatives from plant origin to synthetic and chemical biocides is an increasing major global demand. The antimicrobial activity of eight plant extracts against S. Typhimurium and S. Enteritidis was in vitro determined using both quantitaive and qualitative assays. The most effective plant extracts, i.e. garlic (Allium sativum) leaves and pomegranate (Punica granatum) peels, were applied in laying farms as aerosol, for sanitizing flocks environment, and immersion solution, for disinfecting the experimentally contaminated eggs. The combination between the minimal inhibitory concentrations (MIC) from the two extracts increased their antibacterial potentiality and resulted in a complete inhibition of contaminating S. enterica growth. The application of plant extracts, however, could be recommended as eco-friendly, safe and powerful alternatives to the frequently applied chemical sanitizers and disinfectants against the invasion of enteric Salmonella in egg farms.

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CHARACTERIZATION OF POLYMICROBIAL ON ENDOTRACHEAL TUBES

BIOFILMS

1 1 , Vandecandelaere Ilse Matthijs Nele , Van Nieuwerburgh 2 Filip , Deforce Diete1.2, Nelis Hans1, Depuydt Pieter3, Coenye Tom1 1

Laboratory of Pharmaceutical Microbiology, Ghent University, Harelbekestraat 72, 9000 Ghent, Belgium 2

Laboratory ofPhannaceutical Biotechnology, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium

3

COHABITATION IN ENVIRONMENT.

THE

CYSTIC

FIBROSIS

LUNG

Fothergill JL1, Liu X2, Addison RJ2, Walshaw MJ3, Ledson MJ3, Southern K4, McNamara P4, Brockhurst MA2, Winstanley C1. 1.Institute of Infection and Global Health, University of Liverpool, UK 2. Institute of Integrative Biology, University of Liverpool, UK. 3. Amanda Unit, Liverpool Heart and Chest Hospital, UK. 4. Alder Hey Children’s Hospital, Liverpool, UK.

Department of Intensive Care, Ghent University Hospital, De Pintelaan 185, Ghent, Belgium

Bacteria in the natural environment form complex, interacting communities. This is also

Mechanically ventilated patients often develop ventilator-associated pneumonia (VAP). In fact, the introduction of an endotracheal tube (ET) and its subsequent colonization are important risk factors for the development of VAP. In the present study, we investigated the bacterial diversity of ET biofilms by culture dependent (i.e. selective growth media, 16S rRNA gene sequencing and biochemical tests) and culture independent techniques (i.e. 16S rRNA gene clone libraries and pyrosequencing). In total, 554 isolates were obtained using selective growth media. The majority of the Gram-positive bacteria were identified as Micrococcus luteus and Staphylococcus spp. including Staphylococcus epidermidis, Staphylococcus aureus and Staphylococcus saprophyticus. Also, lactic acid bacteria, corynefonns, Bacillus spp. and Streptomyces spp. were identified. Pseudomonas aeruginosa and Enterobacteriaceae were the dominant groups among the Gram-negative bacteria. In addition, 65 isolates were identified as Candida spp.

true for bacterial infections of animals and humans, where much of the diversity that we see may be

Culture independent identification techniques relied on the construction of 16S rRNA gene clone libraries and pyrosequencing of PCR amplified 16S rRNA genes of selected samples. Sequencing the clone libraries showed that Enterobacteriaceae and lactic acid bacteria were the most dominant groups. In general, sequencing of the clone libraries confirmed the presence of typical aerobic pathogens as observed by the culture dependent identification techniques. In contrast, pyrosequencing of 4 selected samples revealed the presence of typical oral bacteria such as Prevotella spp., Veillonella spp. and lactic acid bacteria.

driven by microbes interacting with each other as well as with the host. The Cystic Fibrosis (CF) lung facilitates the cohabitation of diverse microbial organisms and we are only just beginning to understand the extent of this diversity. In order to characterise the CF lung microbiome, matched sputum and bronchoalveolar lavage (BAL) samples from three children were sequenced using 454 pyrosequencing of 16S rRNA sequences. This enabled the microbial community in each sample to be identified and compared to determine how representative sputum samples are of the lower lung. Furthermore, BAL samples from the left and right lung were also compared to determine spatial differences in the CF lung microbial population. Considerable differences in the microbial population could be found between samples. A retrospective analysis of published data was performed to further describe the CF lung meta-community. This revealed an extremely diverse range of bacteria that are capable colonising the CF lung, with 317 species from 172 different genera identified. These data will be used to develop a multispecies biofilm model in which microbial interactions (both antagonistic and synergistic) can be investigated. Understanding these complex interactions may uncover novel therapeutic targets and ultimately lead to altered CF patient management.

In conclusion, a combination of culture dependent and independent techniques is necessary to obtain a complete picture of the diversity in polymicrobial biofilms present on endotracheal tubes.

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Poster abstracts

PsrP PROMOTES BIOFILM FORMATION BUT INHIBITS ADHESION.

CHILDREN EXPOSURE TO CAMPYLOBACTERIOSIS VIA POULTRY BACKYARDS

Vimberg Vladimir

Wael F. El-Tras1; Ahmed A. Tayel2; Nevein N. El-Kady 3

The nasopharynx is the ecological niche where many commensal bacteria and potential respiratory or invasive pathogens like Streptococcus pneumoniae (pneumococcus) may coexist together in biofilm. Biofilm formation by pneumococcus is required for bacteria persistence in the host nasopharynx and is a first step in development of invasive disease. It was previously demonstrated that pneumococcus PsrP can be an intra-species bacterial adhesin. PsrP is a giant protein of 4776 amino acids that could form a filamentous structure able to span the polysaccharide capsular layer. PsrP is encoded by the most invasive pneumococcal clonal lineages of serotypes 1 and 4.

1

Biofilm formation by pneumococcus wild-type (TIGR4, serotype 4), psrP knockout, psrP overexpression strains and the strain, lacking polysaccharide capsule, were observed after 2, 5, 20 hours of bacteria growth. PsrP was required for biofilm formation but inhibited bacteria initial adhesion to surface. Interestingly, polysaccharide capsule had similar, but stronger, effect on biofilm formation. We visualized the capsule, stained with the capsule specific antibodies, in the biofilm by immunofluorescence microscopy. First, wildtype and psrP overexpression strains were more intensively stained with the capsule antibodies than psrP knockout, suggesting that capsule amount around the cells and PsrP expression is connected. Second, the biofilm matrix of wildtype and psrP overexpression strains contained long fibers, stained with the capsule specific antibodies. The fibers were completely absent around the cells of psrP knockout strain. ELISA and EM confirmed that psrP knockout strain has lower amount of the capsule around the cells than wild-type and psrP overexpression strains. Altogether our results suggest that PsrP is a component of pneumococcus polysaccharide capsule that facilitates biofilm matrix formation. Biofilm matrix may be a favorable place for coexistence of different bacterial species together. We will investigate the role of the PsrP in intra-species biofilms development.

Department of Hygiene and Preventive Medicine (Zoonoses), Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt

Genetic Engineering and Biotechnology Research Institute, Minoufiya University, P. O. box 79/22857, ElSadat City, Egypt 2

Childhood and Motherhood Care Unit, Gharbia Health Management, Ministry of Health, Egypt 3

During August 2010 to January 2012, this study was conducted in five villages located in Northern Egypt, where farmers accustomed to grow backyard poultry for their consumption and/or sales, to investigate the probable zoonotic transmission of Campylobacter sp. to children contacted with poultry backyards. Selection of the examined children and poultry fecal samples was based on the presence of children in contact with live poultry. The study population comprised 128 households owned poultry backyards. The child and convenience backyard samples were collected in the same day for each household and examined for the presence of C. jejuni and C. coli. Among examined cases, 21 children out of 131 (16.03%) had the infection with Campylobacter; Fifteen of them were infected with C. jejuni while another 3 children had C. coli infection and the combined infection was recorded in 3 children. Females were more susceptible for Campylobacteriosis than males; the infection percentage in females was 18.42 while in males was 12.73. The rate of infection in age of > 8 to 13 year was 22.22 % while a significantly lower rate (6 %) was recorded in age range of 4 to ≤ 8 years. the total positive poultry samples for Campylobacter were 78 out of 263 examined samples (29.66%). The percentage of combined infection was 12.5% in households with positive backyards (containing only one positive poultry sample) and infected children. The detection of Campylobacter coinfection, among children and connected poultry, may validate the zoonotic transmission of Campylobacteriosis in Egypt.

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NOTES

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of Sciences Leopoldina for hosting this meeting!

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We would like to thank the German National Academy

Coinfections 2012 Book of Abstracts  

Book of Abstracts, Coinfections 2012

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