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Krzysztof Rakus - One of the best experts on this subject based on the ideXlab platform.
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Using Prokaryotic Mutagenesis and In Vivo Bioluminescent Imaging
2016Co-Authors: Recombinant Cyprinid, Maxime Boutier, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Anca Reschner, Guillaume Fournier, Gavin S Wilkie, Herpesvirus Vaccine, Frédéric FarnirAbstract:Cyprinid Herpesvirus 3 (CyHV 3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candi-date, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vac-cine candidate. The safety and efficacy of this strain were studied using an in vivo biolumi-nescent imaging system (IVIS), qPCR, and histopathological examination, whic
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Cyprinid Herpesvirus 3 : An Archetype of Fish AlloHerpesviruses
Advances in virus research, 2015Co-Authors: Maxime Boutier, David M Stone, Catherine Vancsok, Krzysztof Rakus, Keith Way, Maygane Ronsmans, Joanna Jazowiecka-rakus, Léa Morvan, Ma. Michelle D. Peñaranda, S.j. Van BeurdenAbstract:The order Herpesvirales encompasses viruses that share structural, genetic, and biological properties. However, members of this order infect hosts ranging from molluscs to humans. It is currently divided into three phylogenetically related families. The Alloherpesviridae family contains viruses infecting fish and amphibians. There are 12 alloHerpesviruses described to date, 10 of which infect fish. Over the last decade, Cyprinid Herpesvirus 3 (CyHV-3) infecting common and koi carp has emerged as the archetype of fish alloHerpesviruses. Since its first description in the late 1990s, this virus has induced important economic losses in common and koi carp worldwide. It has also had negative environmental implications by affecting wild carp populations. These negative impacts and the importance of the host species have stimulated studies aimed at developing diagnostic and prophylactic tools. Unexpectedly, the data generated by these applied studies have stimulated interest in CyHV-3 as a model for fundamental research. This review intends to provide a complete overview of the knowledge currently available on CyHV-3.
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rational development of an attenuated recombinant Cyprinid Herpesvirus 3 vaccine using prokaryotic mutagenesis and in vivo bioluminescent imaging
PLOS Pathogens, 2015Co-Authors: Maxime Boutier, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Anca Reschner, Guillaume Fournier, Frédéric Farnir, Gavin S Wilkie, Calixte Bayrou, F. LieffrigAbstract:Cyprinid Herpesvirus 3 (CyHV-3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candidate, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vaccine candidate. The safety and efficacy of this strain were studied using an in vivo bioluminescent imaging system (IVIS), qPCR, and histopathological examination, which demonstrated that it enters fish via skin infection similar to the wild type strain. However, compared to the parental wild type strain, the vaccine candidate replicated at lower levels and spread less efficiently to secondary sites of infection. Transmission experiments allowing water contamination with or without additional physical contact between fish demonstrated that the vaccine candidate has a reduced ability to spread from vaccinated fish to naive sentinel cohabitants. Finally, IVIS analyses demonstrated that the vaccine candidate induces a protective mucosal immune response at the portal of entry. Thus, the present study is the first to report the rational development of a recombinant attenuated vaccine against CyHV-3 for mass vaccination of carp. We also demonstrated the relevance of the CyHV-3 carp model for studying alloHerpesvirus transmission and mucosal immunity in teleost skin.
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Veterinary Research This Provisional
2014Co-Authors: Maygane Ronsmans, F. Lieffrig, Maxime Boutier, Krzysztof Rakus, Frédéric Farnir, Daniel Desmecht, Fabien Ectors, Michaël Vandecan, Charles Mélard, Alain VanderplasschenAbstract:PDF corresponds to the article as it appeared upon acceptance. Fully formatted PDF and full text (HTML) versions will be made available soon. Sensitivity and permissivity of Cyprinus carpio to Cyprinid Herpesvirus 3 during the early stages of its development: importance of the epidermal mucus as an innate immune barrier Veterinary Research 2014, 45:100 doi:10.1186/s13567-014-0100-
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Cyprinid Herpesvirus 3: an interesting virus for applied and fundamental research
Veterinary Research, 2013Co-Authors: Krzysztof Rakus, Maxime Boutier, Catherine Vancsok, Maygane Ronsmans, Joanna Jazowiecka-rakus, Ping Ouyang, Anca Reschner, Alain VanderplasschenAbstract:Cyprinid Herpesvirus 3 (CyHV-3), a member of the family Alloherpesviridae is the causative agent of a lethal, highly contagious and notifiable disease in common and koi carp. The economic importance of common and koi carp industries together with the rapid spread of CyHV-3 worldwide, explain why this virus became soon after its isolation in the 1990s a subject of applied research. In addition to its economic importance, an increasing number of fundamental studies demonstrated that CyHV-3 is an original and interesting subject for fundamental research. In this review, we summarized recent advances in CyHV-3 research with a special interest for studies related to host-virus interactions.
Zenichiro Kawabata - One of the best experts on this subject based on the ideXlab platform.
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effects of daily temperature fluctuation on the survival of carp infected with Cyprinid Herpesvirus 3
Aquaculture, 2014Co-Authors: Teruhiko Takahara, Toshifumi Minamoto, Mie N Honjo, Kimiko Uchii, Hideyuki Doi, Takafumi Ito, Zenichiro KawabataAbstract:Abstract Fluctuating water temperatures can affect fitness in fish when the opportunity to select habitats with appropriate temperature is limited. Despite the importance of the relationships between water temperature and host–pathogen interactions, reports on the susceptibility of fish to infectious viruses under conditions of changing water temperature are limited. Here, we compared the survival rates of common carp (Cyprinus carpio) infected with Cyprinid Herpesvirus 3 (CyHV-3) in water in which the temperature varied from 22 °C ± 3 °C and in water with a constant temperature of 22 °C or 25 °C. We also examined changes in concentrations of CyHV-3 DNA and cortisol released from infected fish into ambient water as indicators of CyHV-3 transmission and stress response, respectively. The survival rates of fish infected with CyHV-3 were lower, and concentrations of CyHV-3 DNA and cortisol were higher, in the fluctuating-temperature treatments than in the constant-temperature treatments. Our findings provide direct evidence that carp are highly susceptible to CyHV-3 infection when water temperatures change diurnally. Moreover, such temperature fluctuations can promote transmission of CyHV-3 in the wild. Preserving a variety of aquatic environments including water temperature may help to prevent disease outbreaks and to conserve fish populations.
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seasonal reactivation enables Cyprinid Herpesvirus 3 to persist in a wild host population
FEMS Microbiology Ecology, 2014Co-Authors: Kimiko Uchii, Toshifumi Minamoto, Mie N Honjo, Zenichiro KawabataAbstract:Emerging infectious diseases are of growing concern in wildlife conservation and animal health. To better understand the consequences of these diseases, a key question lies in how they persist in host populations after they emerge. Using a gene expression approach, we investigated the mechanisms underlying the persistence of an emerging virus, Cyprinid Herpesvirus 3 (CyHV-3), which has been spreading to wild populations of common carp (Cyprinus carpio) in Japan since 2003. Seasonal expression patterns of CyHV-3 genes in wild seropositive carp indicated that replication-related genes were transcribed only during the spring when water temperatures were permissive to CyHV-3 replication. In contrast, possible latency-related genes, which are expressed when CyHV-3 do not multiply, were also transcribed under nonpermissive conditions. These observations suggest that CyHV-3 may persist in carriers by establishing latent infection and then reactivating periodically coincident with the spring temperature increase when carp aggregate for mating, allowing successive virus transmissions between hosts during mating every year. Our results revealed that the life cycle of CyHV-3 may fit perfectly into the ecology of its host, resulting in the long-term persistence of this emerging virus in wild common carp populations.
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nationwide Cyprinid Herpesvirus 3 contamination in natural rivers of japan
Research in Veterinary Science, 2012Co-Authors: Toshifumi Minamoto, Mie N Honjo, Hiroki Yamanaka, Kimiko Uchii, Zenichiro KawabataAbstract:Cyprinid Herpesvirus 3 (CyHV-3) disease is a significant threat for common and koi carp cultivators and for freshwater ecosystems. To determine the prevalence of CyHV-3 in Japanese rivers, a nationwide survey of all national class-A rivers was undertaken in the Summer of 2008. The virus was concentrated from river water samples using the cation-coated filter method. CyHV-3 DNA was detected in 90 rivers, representing 90% of 103 successfully analysed rivers. More than 100,000 copies of CyHV-3 DNA per litre of sample were detected in four rivers, higher than that reported during the Yura River outbreak in 2007. For CyHV-3-positive rivers, the log CyHV-3 density was negatively correlated with the water temperature on the sampling date and positively correlated with the suspended solids and dissolved oxygen, which are annually averaged for each river. Our results demonstrate that virus detection using molecular biology techniques is a powerful tool for monitoring the presence of CyHV-3 in natural environments.
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quantification of Cyprinid Herpesvirus 3 in environmental water by using an external standard virus
Applied and Environmental Microbiology, 2010Co-Authors: Mie N Honjo, Toshifumi Minamoto, Hiroki Yamanaka, Kimiko Uchii, Kazuaki Matsui, Alata A Suzuki, Yukihiro Kohmatsu, Takaji Iida, Zenichiro KawabataAbstract:Cyprinid Herpesvirus 3 (CyHV-3), a lethal DNA virus that spreads in natural lakes and rivers, infects common carp and koi. We established a quantification method for CyHV-3 that includes a viral concentration method and quantitative PCR combined with an external standard virus. Viral concentration methods were compared using the cation-coated filter and ultrafiltration methods. The recovery of virus-like particles was similar for the two methods (cation-coated filter method, 44% ± 19%, n = 3; ultrafiltration method, 50% ± 3%, n = 3); however, the former method was faster and more suitable for routine determinations. The recovery of seeded CyHV-3 based on the cation-coated filter method varied by more than 3 orders of magnitude among the water samples. The recovery yield of CyHV-3 was significantly correlated with that of the seeded λ phage, and the average ratio of λ to the CyHV-3 recovery yield was 1.4, indicating that λ is useful as an external standard virus for determining the recovery yield of CyHV-3. Therefore, to quantify CyHV-3 in environmental water, a known amount of λ was added as an external standard virus to each water sample. Using this method, CyHV-3 DNA was detected in 6 of the 10 (60%) types of environmental water tested; the highest concentration of CyHV-3 DNA was 2 × 105 copies liter−1. The lowest recovery limit of CyHV-3 DNA was 60 copies liter−1. This method is practical for monitoring CyHV-3 abundance in environmental water.
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quantification of Cyprinid Herpesvirus 3 in environmental water by using an external standard virus
Applied and Environmental Microbiology, 2010Co-Authors: Mie N Honjo, Toshifumi Minamoto, Hiroki Yamanaka, Kimiko Uchii, Kazuaki Matsui, Alata A Suzuki, Yukihiro Kohmatsu, Takaji Iida, Zenichiro KawabataAbstract:Cyprinid Herpesvirus 3 (CyHV-3) is a lethal DNA virus that infects the common carp (Cyprinus carpio L.) and koi carp (C. carpio koi). The occurrence of the disease in the United Kingdom has been dated to 1996, following outbreaks in the United States, Israel, Europe, and South Asia (10), and has afflicted cultured ornamental and common carps, causing severe losses to fish breeders, retailers, and hobbyists (28). Therefore, the characterization and diagnosis of the disease have been the subject of intensive research (15). In recent years, the mortality of wild carp has been reported in natural freshwater environments (11, 18, 23). In Lake Biwa in Japan, 60 to 80% of the wild carp population (>100,000) died in 2004, presumably due to CyHV-3 infection (Shiga Prefectural Government, http://www.pref.shiga.jp/g/suisan-s/seika/files/seikah1711.pdf [in Japanese]) (18). The mass mortality of wild carp can directly and indirectly affect community composition and environmental ecosystems (18). Nevertheless, the occurrence of the disease and the means of transmission of CyHV-3 in the natural environment are still not well understood. CyHV-3 is present in several organs of infected fish, such as the intestines, kidneys (7), and gills (29). CyHV-3 is also detected in droppings (3); therefore, infected fish are suspected of releasing CyHV-3 into natural waters. Seasonal variation and the spatial distribution of CyHV-3 may be important for understanding the transmission routes and mechanisms by which CyHV-3 spreads. However, the lack of a reliable method for quantifying CyHV-3 in environmental water precludes our elucidation of how this disease spreads. In general, the concentration of a pathogen in environmental water is considerably lower than that found in host bodies. Therefore, a CyHV-3 concentration method is required to detect and quantify the virus in environmental water. Several methods have been developed for determining concentrations of viruses in water samples. Ultrafiltration can concentrate a pathogen from a large volume of water in <100 liters (27, 35). An alternative method involving the use of electronegative or electropositive microporous adsorbent filters has also been used to concentrate viruses from environmental water (1, 8). The mechanism of concentration in this method is based on electrostatic interactions. Haramoto et al. established a cation-coated filter method in which viruses that had been trapped were eluted with NaOH solution (pH 10.8) instead of the conventional solution, beef extract, which inhibits the PCR (12, 13). The concentrated viruses can then be used for PCR-mediated identification. Using this method, they succeeded in the qualitative detection of CyHV-3 DNA from river water samples (13). Viral recovery during concentration is influenced by soluble organic compounds (33, 34) and salts (31) in the water, which may vary in each sample. Therefore, quantification of the viral DNA from concentrated environmental water samples has been difficult. Because sediments contain many substances that influence DNA recovery, Mumy and Findlay developed a method for the routine determination of DNA extraction efficiency using an external DNA recovery standard, as follows: λ DNA was added to sediments, the total DNA was extracted, and the amount of target DNA recovered was determined by quantitative PCR (22). In this study, we established a method for quantifying CyHV-3 in environmental water using a viral concentration method and TaqMan PCR combined with an external standard virus. To choose a suitable viral concentration method, we compared the viral recovery yields between the ultrafiltration and cation-coated filter methods, and the procedure was modified to increase sensitivity. We then confirmed that the recovery yields of CyHV-3 and the external standard virus λ from different environmental waters throughout the procedure were positively correlated. Finally, we applied this method to environmental water samples taken from Lake Biwa and Takaragaike Pond in Japan at 3 years and 1 month, respectively, after an outbreak of the disease for the detection and quantification of CyHV-3.
Guillaume Fournier - One of the best experts on this subject based on the ideXlab platform.
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Using Prokaryotic Mutagenesis and In Vivo Bioluminescent Imaging
2016Co-Authors: Recombinant Cyprinid, Maxime Boutier, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Anca Reschner, Guillaume Fournier, Gavin S Wilkie, Herpesvirus Vaccine, Frédéric FarnirAbstract:Cyprinid Herpesvirus 3 (CyHV 3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candi-date, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vac-cine candidate. The safety and efficacy of this strain were studied using an in vivo biolumi-nescent imaging system (IVIS), qPCR, and histopathological examination, whic
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rational development of an attenuated recombinant Cyprinid Herpesvirus 3 vaccine using prokaryotic mutagenesis and in vivo bioluminescent imaging
PLOS Pathogens, 2015Co-Authors: Maxime Boutier, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Anca Reschner, Guillaume Fournier, Frédéric Farnir, Gavin S Wilkie, Calixte Bayrou, F. LieffrigAbstract:Cyprinid Herpesvirus 3 (CyHV-3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candidate, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vaccine candidate. The safety and efficacy of this strain were studied using an in vivo bioluminescent imaging system (IVIS), qPCR, and histopathological examination, which demonstrated that it enters fish via skin infection similar to the wild type strain. However, compared to the parental wild type strain, the vaccine candidate replicated at lower levels and spread less efficiently to secondary sites of infection. Transmission experiments allowing water contamination with or without additional physical contact between fish demonstrated that the vaccine candidate has a reduced ability to spread from vaccinated fish to naive sentinel cohabitants. Finally, IVIS analyses demonstrated that the vaccine candidate induces a protective mucosal immune response at the portal of entry. Thus, the present study is the first to report the rational development of a recombinant attenuated vaccine against CyHV-3 for mass vaccination of carp. We also demonstrated the relevance of the CyHV-3 carp model for studying alloHerpesvirus transmission and mucosal immunity in teleost skin.
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Open Access Skin mucus of Cyprinus carpio inhibits Cyprinid Herpesvirus 3 binding to epidermal cells
2013Co-Authors: Victor Stalin Raj, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Benjamin Michel, Guillaume Fournier, Berenice Costes, Cédric Delforges, Frédéric Farnir, Baptiste LeroyAbstract:Skin mucus of Cyprinus carpio inhibits Cyprinid Herpesvirus 3 binding to epidermal cells Raj et al
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Feeding Cyprinus carpio with infectious materials mediates Cyprinid Herpesvirus 3 entry through infection of pharyngeal periodontal mucosa
Veterinary Research, 2012Co-Authors: Guillaume Fournier, F. Lieffrig, Maxime Boutier, Jan Mast, Frédéric Farnir, Victor Stalin Raj, Eric Parmentier, Pierre Vanderwalle, Dominique Peeters, Laurent GilletAbstract:Cyprinid Herpesvirus 3 (CyHV-3), also known as Koi Herpesvirus, is the etiological agent of a mortal disease in common and koi carp. Recently, we investigated the entry of CyHV-3 in carp using bioluminescence imaging and a CyHV-3 recombinant strain expressing luciferase (LUC). We demonstrated that the skin is the major portal of entry after inoculation of carp by immersion in water containing CyHV-3. While this model of infection mimics some natural conditions in which infection takes place, other epidemiological conditions could favour entry of virus through the digestive tract. Here, we investigated whether ingestion of infectious materials mediates CyHV-3 entry through the digestive tract. Carp were fed with materials contaminated with the CyHV-3 LUC recombinant (oral contamination) or immersed in water containing the virus (contamination by immersion). Bioluminescence imaging analyses performed at different times post-infection led to the following observations: (i) the pharyngeal periodontal mucosa is the major portal of entry after oral contamination, while the skin is the major portal of entry after contamination by immersion. (ii) Both modes of inoculation led to the spreading of the infection to the various organs tested. However, the timing and the sequence in which some of the organs turned positive were different between the two modes of inoculation. Finally, we compared the disease induced by the two inoculation modes. They led to comparable clinical signs and mortality rate. The results of the present study suggest that, based on epidemiological conditions, CyHV-3 can enter carp either by skin or periodontal pharyngeal mucosal infection.
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Skin mucus of Cyprinus carpio inhibits Cyprinid Herpesvirus 3 binding to epidermal cells
Veterinary Research, 2011Co-Authors: Victor Stalin Raj, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Benjamin Michel, Guillaume Fournier, Berenice Costes, Cédric Delforges, Frédéric Farnir, Baptiste LeroyAbstract:Cyprinid Herpesvirus 3 (CyHV-3) is the aetiological agent of a mortal and highly contagious disease in common and koi carp. The skin is the major portal of entry of CyHV-3 in carp after immersion in water containing the virus. In the present study, we used in vivo bioluminescence imaging to investigate the effect of skin mucus removal and skin epidermis lesion on CyHV-3 entry. Physical treatments inducing removal of the mucus up to complete erosion of the epidermis were applied on a defined area of carp skin just before inoculation by immersion in infectious water. CyHV-3 entry in carp was drastically enhanced on the area of the skin where the mucus was removed with or without associated epidermal lesion. To investigate whether skin mucus inhibits CyHV-3 binding to epidermal cells, tail fins with an intact mucus layer or without mucus were inoculated ex vivo. While electron microscopy examination revealed numerous viral particles bound on the fins inoculated after mucus removal, no particle could be detected after infection of mucus-covered fins. Finally, anti-CyHV-3 neutralising activity of mucus extract was tested in vitro. Incubation of CyHV-3 with mucus extract reduced its infectivity in a dose dependent manner. The present study demonstrates that skin mucus removal and epidermal lesions enhance CyHV-3 entry in carp. It highlights the role of fish skin mucus as an innate immune protection against viral epidermal entry.
F. Lieffrig - One of the best experts on this subject based on the ideXlab platform.
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rational development of an attenuated recombinant Cyprinid Herpesvirus 3 vaccine using prokaryotic mutagenesis and in vivo bioluminescent imaging
PLOS Pathogens, 2015Co-Authors: Maxime Boutier, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Anca Reschner, Guillaume Fournier, Frédéric Farnir, Gavin S Wilkie, Calixte Bayrou, F. LieffrigAbstract:Cyprinid Herpesvirus 3 (CyHV-3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candidate, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vaccine candidate. The safety and efficacy of this strain were studied using an in vivo bioluminescent imaging system (IVIS), qPCR, and histopathological examination, which demonstrated that it enters fish via skin infection similar to the wild type strain. However, compared to the parental wild type strain, the vaccine candidate replicated at lower levels and spread less efficiently to secondary sites of infection. Transmission experiments allowing water contamination with or without additional physical contact between fish demonstrated that the vaccine candidate has a reduced ability to spread from vaccinated fish to naive sentinel cohabitants. Finally, IVIS analyses demonstrated that the vaccine candidate induces a protective mucosal immune response at the portal of entry. Thus, the present study is the first to report the rational development of a recombinant attenuated vaccine against CyHV-3 for mass vaccination of carp. We also demonstrated the relevance of the CyHV-3 carp model for studying alloHerpesvirus transmission and mucosal immunity in teleost skin.
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Veterinary Research This Provisional
2014Co-Authors: Maygane Ronsmans, F. Lieffrig, Maxime Boutier, Krzysztof Rakus, Frédéric Farnir, Daniel Desmecht, Fabien Ectors, Michaël Vandecan, Charles Mélard, Alain VanderplasschenAbstract:PDF corresponds to the article as it appeared upon acceptance. Fully formatted PDF and full text (HTML) versions will be made available soon. Sensitivity and permissivity of Cyprinus carpio to Cyprinid Herpesvirus 3 during the early stages of its development: importance of the epidermal mucus as an innate immune barrier Veterinary Research 2014, 45:100 doi:10.1186/s13567-014-0100-
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Feeding Cyprinus carpio with infectious materials mediates Cyprinid Herpesvirus 3 entry through infection of pharyngeal periodontal mucosa
Veterinary Research, 2012Co-Authors: Guillaume Fournier, F. Lieffrig, Maxime Boutier, Jan Mast, Frédéric Farnir, Victor Stalin Raj, Eric Parmentier, Pierre Vanderwalle, Dominique Peeters, Laurent GilletAbstract:Cyprinid Herpesvirus 3 (CyHV-3), also known as Koi Herpesvirus, is the etiological agent of a mortal disease in common and koi carp. Recently, we investigated the entry of CyHV-3 in carp using bioluminescence imaging and a CyHV-3 recombinant strain expressing luciferase (LUC). We demonstrated that the skin is the major portal of entry after inoculation of carp by immersion in water containing CyHV-3. While this model of infection mimics some natural conditions in which infection takes place, other epidemiological conditions could favour entry of virus through the digestive tract. Here, we investigated whether ingestion of infectious materials mediates CyHV-3 entry through the digestive tract. Carp were fed with materials contaminated with the CyHV-3 LUC recombinant (oral contamination) or immersed in water containing the virus (contamination by immersion). Bioluminescence imaging analyses performed at different times post-infection led to the following observations: (i) the pharyngeal periodontal mucosa is the major portal of entry after oral contamination, while the skin is the major portal of entry after contamination by immersion. (ii) Both modes of inoculation led to the spreading of the infection to the various organs tested. However, the timing and the sequence in which some of the organs turned positive were different between the two modes of inoculation. Finally, we compared the disease induced by the two inoculation modes. They led to comparable clinical signs and mortality rate. The results of the present study suggest that, based on epidemiological conditions, CyHV-3 can enter carp either by skin or periodontal pharyngeal mucosal infection.
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Cyprinid Herpesvirus 3
Emerging Infectious Diseases, 2010Co-Authors: Benjamin Michel, F. Lieffrig, Guillaume Fournier, Berenice Costes, Alain VanderplasschenAbstract:The recently designated Cyprinid Herpesvirus 3 (CyHV-3) is an emerging agent that causes fatal disease in common and koi carp. Since its emergence in the late 1990s, this highly contagious pathogen has caused severe financial losses in common and koi carp culture industries worldwide. In addition to its economic role, recent studies suggest that CyHV-3 may have a role in fundamental research. CyHV-3 has the largest genome among viruses in the order Herpesvirales and serves as a model for mutagenesis of large DNA viruses. Other studies suggest that the skin of teleost fish represents an efficient portal of entry for certain viruses. The effect of temperature on viral replication suggests that the body temperature of its poikilotherm host could regulate the outcome of the infection (replicative vs. nonreplicative). Recent advances with regard to CyHV-3 provide a role for this virus in fundamental and applied research.
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the genome of Cyprinid Herpesvirus 3 encodes 40 proteins incorporated in mature virions
Journal of General Virology, 2010Co-Authors: Benjamin Michel, F. Lieffrig, Baptiste Leroy, Ruddy Wattiez, Alain Vanderplasschen, Stalin V Raj, Jan Mast, Berenice CostesAbstract:Koi Herpesvirus, also known as Cyprinid Herpesvirus 3 (CyHV-3), is the aetiological agent of an emerging and mortal disease in common and koi carp. CyHV-3 virions present the characteristic morphology of other members of the order Herpesvirales, being composed of an envelope, a capsid containing the genome and a tegument. This study identified CyHV-3 structural proteins and the corresponding encoding genes using liquid chromatography tandem mass spectrometry-based proteomic approaches. In addition, exponentially modified protein abundance index analyses were used to estimate the relative abundance of the identified proteins in CyHV-3 virions. These analyses resulted in the identification of 40 structural proteins, which were classified based on bioinformatic analyses as capsid (three), envelope (13), tegument (two) and unclassified (22) structural proteins. Finally, a search for host proteins in purified CyHV-3 virions indicated the potential incorporation of up to 18 distinct cellular proteins. The identification of the proteins incorporated into CyHV-3 virions and determination of the viral genes encoding these proteins are key milestones for further fundamental and applied research on this virus.
Baptiste Leroy - One of the best experts on this subject based on the ideXlab platform.
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Open Access Skin mucus of Cyprinus carpio inhibits Cyprinid Herpesvirus 3 binding to epidermal cells
2013Co-Authors: Victor Stalin Raj, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Benjamin Michel, Guillaume Fournier, Berenice Costes, Cédric Delforges, Frédéric Farnir, Baptiste LeroyAbstract:Skin mucus of Cyprinus carpio inhibits Cyprinid Herpesvirus 3 binding to epidermal cells Raj et al
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Open Access Identification and localization of the structural proteins of anguillid Herpesvirus 1
2013Co-Authors: Baptiste Leroy, Olga L M Haenen, Ben P H Peeters, Peter J M Rottier, Marc Y Engelsma, Ruddy Wattiez, Sjef Boeren, Jacques Jm Vervoort, Alain FAbstract:Many of the known fish Herpesviruses have important aquaculture species as their natural host, and may cause serious disease and mortality. Anguillid Herpesvirus 1 (AngHV-1) causes a hemorrhagic disease in European eel, Anguilla anguilla. Despite their importance, fundamental molecular knowledge on fish Herpesviruses is still limited. In this study we describe the identification and localization of the structural proteins of AngHV-1. Purified virions were fractionated into a capsid-tegument and an envelope fraction, and premature capsids were isolated from infected cells. Proteins were extracted by different methods and identified by mass spectrometry. A total of 40 structural proteins were identified, of which 7 could be assigned to the capsid, 11 to the envelope, and 22 to the tegument. The identification and localization of these proteins allowed functional predictions. Our findings include the identification of the putative capsid triplex protein 1, the predominant tegument protein, and the major antigenic envelope proteins. Eighteen of the 40 AngHV-1 structural proteins had sequence homologues in related Cyprinid Herpesvirus 3 (CyHV-3). Conservation of fish Herpesvirus structural genes seemed to be high for the capsid proteins, limited for the tegument proteins, and low for the envelope proteins. The identification and localization of the structural proteins of AngHV-1 in this study adds to the fundamental knowledge of members of the Alloherpesviridae family, especially of the Cyprinivirus genus
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Identification and localization of the structural proteins of anguillid Herpesvirus 1
Veterinary Research, 2011Co-Authors: Steven J Van Beurden, Olga L M Haenen, Ben P H Peeters, Peter J M Rottier, Marc Y Engelsma, Baptiste Leroy, Ruddy Wattiez, Sjef Boeren, Jacques Jm Vervoort, Alain VanderplasschenAbstract:Many of the known fish Herpesviruses have important aquaculture species as their natural host, and may cause serious disease and mortality. Anguillid Herpesvirus 1 (AngHV-1) causes a hemorrhagic disease in European eel, Anguilla anguilla . Despite their importance, fundamental molecular knowledge on fish Herpesviruses is still limited. In this study we describe the identification and localization of the structural proteins of AngHV-1. Purified virions were fractionated into a capsid-tegument and an envelope fraction, and premature capsids were isolated from infected cells. Proteins were extracted by different methods and identified by mass spectrometry. A total of 40 structural proteins were identified, of which 7 could be assigned to the capsid, 11 to the envelope, and 22 to the tegument. The identification and localization of these proteins allowed functional predictions. Our findings include the identification of the putative capsid triplex protein 1, the predominant tegument protein, and the major antigenic envelope proteins. Eighteen of the 40 AngHV-1 structural proteins had sequence homologues in related Cyprinid Herpesvirus 3 (CyHV-3). Conservation of fish Herpesvirus structural genes seemed to be high for the capsid proteins, limited for the tegument proteins, and low for the envelope proteins. The identification and localization of the structural proteins of AngHV-1 in this study adds to the fundamental knowledge of members of the Alloherpesviridae family, especially of the Cyprinivirus genus.
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Skin mucus of Cyprinus carpio inhibits Cyprinid Herpesvirus 3 binding to epidermal cells
Veterinary Research, 2011Co-Authors: Victor Stalin Raj, Krzysztof Rakus, Maygane Ronsmans, Ping Ouyang, Benjamin Michel, Guillaume Fournier, Berenice Costes, Cédric Delforges, Frédéric Farnir, Baptiste LeroyAbstract:Cyprinid Herpesvirus 3 (CyHV-3) is the aetiological agent of a mortal and highly contagious disease in common and koi carp. The skin is the major portal of entry of CyHV-3 in carp after immersion in water containing the virus. In the present study, we used in vivo bioluminescence imaging to investigate the effect of skin mucus removal and skin epidermis lesion on CyHV-3 entry. Physical treatments inducing removal of the mucus up to complete erosion of the epidermis were applied on a defined area of carp skin just before inoculation by immersion in infectious water. CyHV-3 entry in carp was drastically enhanced on the area of the skin where the mucus was removed with or without associated epidermal lesion. To investigate whether skin mucus inhibits CyHV-3 binding to epidermal cells, tail fins with an intact mucus layer or without mucus were inoculated ex vivo. While electron microscopy examination revealed numerous viral particles bound on the fins inoculated after mucus removal, no particle could be detected after infection of mucus-covered fins. Finally, anti-CyHV-3 neutralising activity of mucus extract was tested in vitro. Incubation of CyHV-3 with mucus extract reduced its infectivity in a dose dependent manner. The present study demonstrates that skin mucus removal and epidermal lesions enhance CyHV-3 entry in carp. It highlights the role of fish skin mucus as an innate immune protection against viral epidermal entry.
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the genome of Cyprinid Herpesvirus 3 encodes 40 proteins incorporated in mature virions
Journal of General Virology, 2010Co-Authors: Benjamin Michel, F. Lieffrig, Baptiste Leroy, Ruddy Wattiez, Alain Vanderplasschen, Stalin V Raj, Jan Mast, Berenice CostesAbstract:Koi Herpesvirus, also known as Cyprinid Herpesvirus 3 (CyHV-3), is the aetiological agent of an emerging and mortal disease in common and koi carp. CyHV-3 virions present the characteristic morphology of other members of the order Herpesvirales, being composed of an envelope, a capsid containing the genome and a tegument. This study identified CyHV-3 structural proteins and the corresponding encoding genes using liquid chromatography tandem mass spectrometry-based proteomic approaches. In addition, exponentially modified protein abundance index analyses were used to estimate the relative abundance of the identified proteins in CyHV-3 virions. These analyses resulted in the identification of 40 structural proteins, which were classified based on bioinformatic analyses as capsid (three), envelope (13), tegument (two) and unclassified (22) structural proteins. Finally, a search for host proteins in purified CyHV-3 virions indicated the potential incorporation of up to 18 distinct cellular proteins. The identification of the proteins incorporated into CyHV-3 virions and determination of the viral genes encoding these proteins are key milestones for further fundamental and applied research on this virus.
