• 1. International Committee on Taxonomy of Viruses. Virus Taxonomy. Available at: www.ictvonline.org/virusTaxonomy.asp?taxnode_id=20140756. Accessed Mar 8, 2016.

    • Search Google Scholar
    • Export Citation
  • 2. Fölsch DW, Leloup P. Fatale endemische Infektion in einem Serpentarium. Diagnose, Behandlung und prophylaktische Maßnahmen. Tierarztl Prax 1976; 4: 527536.

    • Search Google Scholar
    • Export Citation
  • 3. Jacobson ER, Gaskin JM, Simpson CF, et al. Paramyxo-like virus infection in a rock rattlesnake. J Am Vet Med Assoc 1980; 177: 796799.

    • Search Google Scholar
    • Export Citation
  • 4. Jacobson ER, Gaskin JM, Page D, et al. Illness associated with paramyxo-like virus infection in a zoologic collection of snakes. J Am Vet Med Assoc 1981; 179: 12271230.

    • Search Google Scholar
    • Export Citation
  • 5. Jacobson ER, Gaskin JM, Wells S, et al. Epizootic of ophidian paramyxovirus in a zoological collection: pathological, microbiological and serological findings. J Zoo Wildl Med 1992; 23: 318327.

    • Search Google Scholar
    • Export Citation
  • 6. Kolesnikovas CKM, Grego KF, Rameh de Albuquerque LC, et al. Ophidian paramyxovirus in Brazilian vipers (Bothrops alternatus). Vet Rec 2006; 159: 390392.

    • Search Google Scholar
    • Export Citation
  • 7. Jacobson ER. Viruses and viral diseases of reptiles. In: Jacobson ER, ed. Infectious diseases and pathology of reptiles. Color atlas and text. Boca Raton, Fla: CRC Press, 2007; 395460.

    • Search Google Scholar
    • Export Citation
  • 8. Chitty J. Respiratory system. In: Girling SJ, Raiti P, eds. BSAVA manual of reptiles. 2nd ed. Gloucester, England: British Small Animal Veterinary Association, 2004; 230242.

    • Search Google Scholar
    • Export Citation
  • 9. Hyndman TH, Shilton CM, Marschang RE. Paramyxoviruses in reptiles: a review. Vet Microbiol 2013; 165: 200213.

  • 10. Essbauer S, Ahne W. Viruses of lower vertebrates. J Vet Med B Infect Dis Vet Public Health 2001; 48: 403475.

  • 11. Marschang RE. Viruses infecting reptiles. Viruses 2011; 3: 20872126.

  • 12. Strik NI, Alleman AR, Harr KE. Circulating inflammatory cells. In: Jacobson ER, ed. Infectious diseases and pathology of reptiles. Color atlas and text. Boca Raton, Fla: CRC Press, 2007; 167218.

    • Search Google Scholar
    • Export Citation
  • 13. Zimmerman LM, Vogel LA, Bowden RM. Understanding the vertebrate immune system: insights from the reptilian perspective. J Exp Biol 2010; 213: 661671.

    • Search Google Scholar
    • Export Citation
  • 14. Origgi FC. Reptile immunology. In: Jacobson ER, ed. Infectious diseases and pathology of reptiles. Color atlas and text. Boca Raton, Fla: CRC Press, 2007; 131166.

    • Search Google Scholar
    • Export Citation
  • 15. Merchant ME, Roche CM, Thibodeaux D, et al. Identification of alternative pathway serum complement activity in the blood of the American alligator (Alligator mississippiensis). Comp Biochem Physiol B Biochem Mol Biol 2005; 141: 281288.

    • Search Google Scholar
    • Export Citation
  • 16. Coico R, Sunshine G, eds. Immunology: a short course. 6th ed. Hoboken, NJ: John Wiley & Sons, Inc, 2009.

  • 17. el Ridi R, Wahby AF, Saad AH, et al. Concanavalin A responsiveness and interleukin 2 production in the snake Spalerosophis diadema. Immunobiology 1987; 174: 177189.

    • Search Google Scholar
    • Export Citation
  • 18. Zimmerman LM, Paitz RT, Vogel LA, et al. Variation in the seasonal patterns of innate and adaptive immunity in the red-eared slider (Trachemys scripta). J Exp Biol 2010; 213: 14771483.

    • Search Google Scholar
    • Export Citation
  • 19. Schwanz L, Warner DA, McGaugh S, et al. State-dependent physiological maintenance in a long-lived ectotherm, the painted turtle (Chrysemys picta). J Exp Biol 2011; 214: 8897.

    • Search Google Scholar
    • Export Citation
  • 20. Ambrosius H. Immunoglobulins and antibody production in reptiles. In: Marchalonis JJ, ed. Comparative immunology. Oxford, England: Blackwell Scientific Publications, 1976; 298334.

    • Search Google Scholar
    • Export Citation
  • 21. el Ridi R, Mansour MH, Zada S. Immunoglobulins of the snake Psammophis sibilans: Studies using a monoclonal antibody. Immunobiology 1991; 184: 113.

    • Search Google Scholar
    • Export Citation
  • 22. Work TM, Balazs GH, Rameyer RA, et al. Assessing humoral and cell-mediated immune response in Hawaiian green turtles, Chelonia mydas. Vet Immunol Immunopathol 2000; 74: 179194.

    • Search Google Scholar
    • Export Citation
  • 23. Origgi FC, Klein PA, Mathes K, et al. Enzyme-linked immunosorbent assay for detecting herpesvirus exposure in Mediterranean tortoises (spur-thighed tortoise [Testudo graeca] and Hermann's tortoise [Testudo hermanni]). J Clin Microbiol 2001; 39: 31563163.

    • Search Google Scholar
    • Export Citation
  • 24. Zhou X, Guo Q, Dai H. Molecular characterization and expression profiles in response to bacterial infection of Chinese soft-shelled turtle interleukin-8 (IL-8), the first reptilian chemokine gene. Dev Comp Immunol 2009; 33: 838847.

    • Search Google Scholar
    • Export Citation
  • 25. Longenecker BM, Mosmann TR. “Natural” antibodies to chicken MHC antigens are present in mice, rats, humans, alligators and allogeneic chickens. Immunogenetics 1980; 11: 293302.

    • Search Google Scholar
    • Export Citation
  • 26. Madsen T, Ujvari B, Nandakumar KS, et al. Do “infectious” prey select for high levels of natural antibodies in tropical pythons? Evol Ecol 2007; 21: 271279.

    • Search Google Scholar
    • Export Citation
  • 27. Sparkman AM, Palacios MG. A test of life-history theories of immune defence in two ecotypes of the garter snake, Thamnophis elegans. J Anim Ecol 2009; 78: 12421248.

    • Search Google Scholar
    • Export Citation
  • 28. Gravendyck M, Ammermann P, Marschang RE, et al. Paramyxoviral and reoviral infections of iguanas on Honduran Islands. J Wildl Dis 1998; 34: 3338.

    • Search Google Scholar
    • Export Citation
  • 29. Pees M, Schmidt V, Marschang RE, et al. Prevalence of viral infections in captive collections of boid snakes in Germany. Vet Rec 2010; 166: 422425.

    • Search Google Scholar
    • Export Citation
  • 30. Jacobson ER, Origgi FC. Serodiagnostics. In: Jacobson ER, ed. Infectious diseases and pathology of reptiles. Color atlas and text. Boca Raton, Fla: CRC Press, 2007; 381394.

    • Search Google Scholar
    • Export Citation
  • 31. Herbst LH, Klein PA. Monoclonal antibodies for the measurement of class-specific antibody responses in the green turtle, Chelonia mydas. Vet Immunol Immunopathol 1995; 46: 317335.

    • Search Google Scholar
    • Export Citation
  • 32. Kania SA, Kennedy M, Nowlin N, et al. Development of an enzyme linked immunosorbent assay for the diagnosis of ophidian paramyxovirus, in Proceedings. Eur Assoc Zoo Wildl Vet 3rd Sci Meet 2000; 7173.

    • Search Google Scholar
    • Export Citation
  • 33. Graczyk TK, Cranfield MR. Detection of Cryptosporidium- specific serum immunoglobulins in captive snakes by a polyclonal antibody in the indirect ELISA. Vet Res 1997; 28: 131142.

    • Search Google Scholar
    • Export Citation
  • 34. Lock BA, Green LG, Jacobson ER, et al. Use of an ELISA for detection of antibody responses in Argentine boa constrictors (Boa constrictor occidentalis). Am J Vet Res 2003; 64: 388395.

    • Search Google Scholar
    • Export Citation
  • 35. Jacobson ER, Adams HP, Geisbert TW, et al. Pulmonary lesions in experimental ophidian paramyxovirus pneumonia of Aruba island rattlesnakes, Crotalus unicolor. Vet Pathol 1997; 34: 450459.

    • Search Google Scholar
    • Export Citation
  • 36. Jacobson ER, Gaskin JM, Flanagan JP, et al. Antibody responses of Western diamondback rattlesnakes (Crotalus atrox) to inactivated ophidian Paramyxovirus vaccines. J Zoo Wildl Med 1991; 22: 184190.

    • Search Google Scholar
    • Export Citation
  • 37. Pees M, Schmidt V, Schlömer J, et al. Significance of the sampling points and the aerobic microbiological culture for the diagnosis of respiratory infections in reptiles. Dtsch Tierarztl Wochenschr 2007; 114: 388393.

    • Search Google Scholar
    • Export Citation
  • 38. Schmidt V, Marschang RE, Abbas MD, et al. Detection of pathogens in Boidae and Pythonidae with and without respiratory disease. Vet Rec 2013; 172: 236.

    • Search Google Scholar
    • Export Citation
  • 39. Ahne W, Batts WN, Kurath G, et al. Comparative sequence analyses of sixteen reptilian paramyxoviruses. Virus Res 1999; 63: 6574.

  • 40. Pees M, Neul A, Müller K, et al. Virus distribution and detection in corn snakes (Pantherophis guttatus) after experimental infection with three different ferlavirus strains. Vet Microbiol 2016; 182: 213222.

    • Search Google Scholar
    • Export Citation
  • 41. Marschang RE, Papp T, Frost JW. Comparison of paramyxovirus isolates from snakes, lizards and a tortoise. Virus Res 2009; 144: 272279.

    • Search Google Scholar
    • Export Citation
  • 42. Abbas MD, Marschang RE, Schmidt V, et al. A unique novel reptilian paramyxovirus, four atadenovirus types and a reovirus identified in a concurrent infection of a corn snake (Pantherophis guttatus) collection in Germany. Vet Microbiol 2011; 150: 7079.

    • Search Google Scholar
    • Export Citation
  • 43. Hernandez-Divers SJ. Diagnostic techniques. In: Mader DR, ed. Reptile medicine and surgery. 2nd ed. St Louis: Saunders Elsevier, 2006; 490532.

    • Search Google Scholar
    • Export Citation
  • 44. Clark P, Boardman W, Raidal S, eds. Atlas of clinical avian hematology. New York: Wiley-Blackwell, 2009; 132.

  • 45. Campbell TW, Ellis CK, eds. Avian and exotic animal hematology and cytology. 3rd ed. Ames, Iowa: Blackwell Publishing, 2007; 381.

  • 46. Fudge AM, ed. Laboratory medicine: avian and exotic pets. Philadelphia: Saunders Co, 2000; 1264.

  • 47. Rösler R, Abbas MD, Papp T, et al. Detection of antibodies against paramyxoviruses in tortoises. J Zoo Wildl Med 2013; 44: 333339.

    • Search Google Scholar
    • Export Citation
  • 48. Wilson MB, Nakane PK. Recent developments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies. In: Knapp W, Holubar K, Wick G, eds. Immunofluorescence and related straining techniques. Amsterdam: Elsevier/North Holland Biomedical Press, 1978; 215224.

    • Search Google Scholar
    • Export Citation
  • 49. Mukaka MM. Statistics corner: a guide to appropriate use of correlation coefficient in medical research. Malawi Med J 2012; 24: 6971.

    • Search Google Scholar
    • Export Citation
  • 50. Eze CP, Okoye JOA, Ogbonna IO, et al. Comparative evaluation of the effects of velogenic Newcastle disease virus infection on the hematology of ducks and chickens. Open J Vet Med 2014; 4: 113121.

    • Search Google Scholar
    • Export Citation
  • 51. Orós J, Sicilia J, Torrent A, et al. Immunohistochemical detection of ophidian paramyxovirus in snakes in the Canary Islands. Vet Rec 2001; 149: 2123.

    • Search Google Scholar
    • Export Citation
  • 52. Allender MC, Mitchell MA, Dreslik MJ, et al. Measuring agreement and discord among hemagglutination inhibition assays against different ophidian paramyxovirus strains in the Eastern massasauga (Sistrurus catenatus catenatus). J Zoo Wildl Med 2008; 39: 358361.

    • Search Google Scholar
    • Export Citation
  • 53. Papp T, Seybold J, Marschang RE. Paramyxovirus infection in a leopard tortoise (Geochelone pardalis babcocki) with respiratory disease. J Herpetol Med Surg 2010; 20: 6468.

    • Search Google Scholar
    • Export Citation
  • 54. Hellebuyck T, Van Steendam K, Deforce D, et al. Autovaccination confers protection against Devriesea agamarum associated septicemia but not dermatitis in bearded dragons (Pogona vitticeps). PLoS One 2014; 9: e113084.

    • Search Google Scholar
    • Export Citation
  • 55. Jacobson ER, Origgi FC. Use of serology in reptile medicine. Sem Avian Exot Pet Med 2002; 11: 3345.

  • 56. Pettinello R, Dooley H. The immunoglobulins of cold-blooded vertebrates. Biomolecules 2014; 4: 10451069.

  • 57. Woodruff JF, Woodruff JJ. The effect of viral infections on the function of the immune system. In: Notkins AL, ed. Viral immunology and immunopathology. New York: Academic Press, 1975; 393418.

    • Search Google Scholar
    • Export Citation
  • 58. Dein FJ, Wilson A, Fischer D, et al. Avian leucocyte counting using the hemocytometer. J Zoo Wildl Med 1994; 25: 432437.

  • 59. Tavares-Dias M. Oliveira-Júnior, Marcon JL. Methodological limitations of counting total leukocytes and thrombocytes in reptiles (Amazon turtle, Podocnemis expansa): an analysis and discussion. Acta Amazon 2008; 38: 351356.

    • Search Google Scholar
    • Export Citation

Advertisement

Immunologic responses in corn snakes (Pantherophis guttatus) after experimentally induced infection with ferlaviruses

Annkatrin Neul Med Vet1, Wieland Schrödl PD, Dr Med Vet2, Rachel E. Marschang PD, Dr Med Vet3, Tina Bjick Med Vet4, Uwe Truyen Prof Dr Med Vet5, Heiner von Buttlar Dr Med Vet6, and Michael Pees Prof Dr Med Vet7
View More View Less
  • 1 Clinic for Birds and Reptiles, University of Leipzig, 04103 Leipzig, Germany.
  • | 2 Institute of Bacteriology and Mycology, University of Leipzig, 04103 Leipzig, Germany.
  • | 3 Laboklin GmbH & Co KG, Steubenstraße 4, 97688 Bad Kissingen, Germany.
  • | 4 Clinic for Birds and Reptiles, University of Leipzig, 04103 Leipzig, Germany.
  • | 5 Institute of Animal Hygiene and Veterinary Public Health, University of Leipzig, 04103 Leipzig, Germany.
  • | 6 Institute of Immunology, University of Leipzig, 04103 Leipzig, Germany.
  • | 7 Clinic for Birds and Reptiles, University of Leipzig, 04103 Leipzig, Germany.

Abstract

OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses.

ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes.

PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days −6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed.

RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses.

CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.

Abstract

OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses.

ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes.

PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days −6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed.

RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses.

CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.

Supplementary Materials

    • Supplementary Table 1 (PDF 120 kb)
    • Supplementary Table 2 (PDF 153 kb)
    • Supplementary Table 3 (PDF 97 kb)
    • Supplementary Table 4 (PDF 91 kb)

Contributor Notes

Dr. von Buttlar's present address is Bundeswehr Institute of Microbiology, Department of Bacteriology and Toxinology, Neuherbergstrasse 11, 80937 Munich, Germany.

Address correspondence to Dr. Pees (pees@vogelklinik.uni-leipzig.de).