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Feeding Cyprinus carpio with infectious materials mediates cyprinid herpesvirus 3 entry through infection of pharyngeal periodontal mucosa.

Fournier G, Boutier M, Stalin Raj V, Mast J, Parmentier E, Vanderwalle P, Peeters D, Lieffrig F, Farnir F, Gillet L, Vanderplasschen A - Vet. Res. (2012)

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Immunology-Vaccinology (B43b), Department of Infectious and Parasitic Diseases (B43b), Faculty of Veterinary Medicine, University of Liège, 4000 Liège, Belgium. a.vdplasschen@ulg.ac.be.

ABSTRACT
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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Progression of CyHV-3 infection in carp analysed by bioluminescence imaging. Two groups of fish (mean weight of 10 g) were infected with the CyHV-3 LUC strain either by bathing them in water containing the virus (Immersion, left column) or by feeding them with food pellets contaminated with the virus (Oral, right column). At the indicated time post-infection, six fish per group were analysed by in vivo and ex vivo bioluminescence imaging. A/For each fish, the IVIS signal (Photon/second) was determined for several organs (skin, gills, pharynx, pseudogaster, intestine, and heart) as described in the materials and methods. B/For each analysed organ, the number of positive fish is presented according to time post-infection. This experiment is representative of two independent experiments.
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Figure 4: Progression of CyHV-3 infection in carp analysed by bioluminescence imaging. Two groups of fish (mean weight of 10 g) were infected with the CyHV-3 LUC strain either by bathing them in water containing the virus (Immersion, left column) or by feeding them with food pellets contaminated with the virus (Oral, right column). At the indicated time post-infection, six fish per group were analysed by in vivo and ex vivo bioluminescence imaging. A/For each fish, the IVIS signal (Photon/second) was determined for several organs (skin, gills, pharynx, pseudogaster, intestine, and heart) as described in the materials and methods. B/For each analysed organ, the number of positive fish is presented according to time post-infection. This experiment is representative of two independent experiments.

Mentions: Imaging of firefly (Photinus pyralis) LUC was performed using an "in vivo imaging system" (IVIS) (IVIS®spectrum, Xenogen, Caliper LifeSciences, Hopkinton, Massachusetts, USA) as described previously [10]. Fish were anesthetized with benzocaine (50 mg/L of water). Fifteen minutes before bioluminescence analysis, D-luciferin (150 mg/kg body weight) (Xenogen, Caliper LifeSciences, Hopkinton, Massachusetts, USA) was administered by intraperitoneal injection. Each fish was analysed in vivo lying on its right and its left side and ex vivo after euthanasia and dissection. All the images presented in this study were acquired using a field view of 15 cm, an auto-exposure time with a maximum of 1 minute, a binning factor of 4 and a f/stop of 1. Relative intensities of transmitted light from bioluminescence were represented as a pseudocolor image ranging from violet (least intense) to red (most intense). Corresponding grey-scale photographs and color luciferase images were superimposed using the LivingImage analysis software (Xenogen, Caliper LifeSciences, Hopkinton, Massachusetts, USA). For quantitative comparisons, the Living Image software (Caliper Life Sciences) was used to obtain the total flux (p.s-1) over each region of interest (ROI). All the ROI automatically identified by the IVIS software as positive (Figure 4a) were standing out against background with a difference of at least 3 log.


Feeding Cyprinus carpio with infectious materials mediates cyprinid herpesvirus 3 entry through infection of pharyngeal periodontal mucosa.

Fournier G, Boutier M, Stalin Raj V, Mast J, Parmentier E, Vanderwalle P, Peeters D, Lieffrig F, Farnir F, Gillet L, Vanderplasschen A - Vet. Res. (2012)

Progression of CyHV-3 infection in carp analysed by bioluminescence imaging. Two groups of fish (mean weight of 10 g) were infected with the CyHV-3 LUC strain either by bathing them in water containing the virus (Immersion, left column) or by feeding them with food pellets contaminated with the virus (Oral, right column). At the indicated time post-infection, six fish per group were analysed by in vivo and ex vivo bioluminescence imaging. A/For each fish, the IVIS signal (Photon/second) was determined for several organs (skin, gills, pharynx, pseudogaster, intestine, and heart) as described in the materials and methods. B/For each analysed organ, the number of positive fish is presented according to time post-infection. This experiment is representative of two independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3295659&req=5

Figure 4: Progression of CyHV-3 infection in carp analysed by bioluminescence imaging. Two groups of fish (mean weight of 10 g) were infected with the CyHV-3 LUC strain either by bathing them in water containing the virus (Immersion, left column) or by feeding them with food pellets contaminated with the virus (Oral, right column). At the indicated time post-infection, six fish per group were analysed by in vivo and ex vivo bioluminescence imaging. A/For each fish, the IVIS signal (Photon/second) was determined for several organs (skin, gills, pharynx, pseudogaster, intestine, and heart) as described in the materials and methods. B/For each analysed organ, the number of positive fish is presented according to time post-infection. This experiment is representative of two independent experiments.
Mentions: Imaging of firefly (Photinus pyralis) LUC was performed using an "in vivo imaging system" (IVIS) (IVIS®spectrum, Xenogen, Caliper LifeSciences, Hopkinton, Massachusetts, USA) as described previously [10]. Fish were anesthetized with benzocaine (50 mg/L of water). Fifteen minutes before bioluminescence analysis, D-luciferin (150 mg/kg body weight) (Xenogen, Caliper LifeSciences, Hopkinton, Massachusetts, USA) was administered by intraperitoneal injection. Each fish was analysed in vivo lying on its right and its left side and ex vivo after euthanasia and dissection. All the images presented in this study were acquired using a field view of 15 cm, an auto-exposure time with a maximum of 1 minute, a binning factor of 4 and a f/stop of 1. Relative intensities of transmitted light from bioluminescence were represented as a pseudocolor image ranging from violet (least intense) to red (most intense). Corresponding grey-scale photographs and color luciferase images were superimposed using the LivingImage analysis software (Xenogen, Caliper LifeSciences, Hopkinton, Massachusetts, USA). For quantitative comparisons, the Living Image software (Caliper Life Sciences) was used to obtain the total flux (p.s-1) over each region of interest (ROI). All the ROI automatically identified by the IVIS software as positive (Figure 4a) were standing out against background with a difference of at least 3 log.

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Immunology-Vaccinology (B43b), Department of Infectious and Parasitic Diseases (B43b), Faculty of Veterinary Medicine, University of Liège, 4000 Liège, Belgium. a.vdplasschen@ulg.ac.be.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus