2004, Number 1
Next >>
Vet Mex 2004; 35 (1)
Avian influenza: histopathology and viral detection in formalin-fixed, paraffin-embedded tissues by RT-PCR
Padilla NR, Aburto FE, Fraire CM, Padilla NL
Language: English/Spanish
References: 33
Page: 1-18
PDF size: 604.45 Kb.
ABSTRACT
The aim of this study was to establish a reverse transcription-polymerase chain
reaction (RT-PCR) for the diagnosis of avian influenza virus in paraffin embedded
tissues of experimentally infected animals, and to correlate viral detection
by this method with the histological changes. Eight groups of 10 Arbor Acres
chickens of 5 weeks of age were inoculated intravenously with 101 to 108 50%
chicken lethal doses of avian influenza virus A/Ck/Queretaro/20/95 (H5N2). All
chickens were subjected to a general histopathological examination. Three organs
were analyzed by RT-PCR (one chicken per dose): lung, encephalon, and kidney.
The main histopathologic lesions in all organs examined from infected birds
were: necrosis of lymphoid tissue (in lung, spleen, bursa of Fabricius, and
cecal tonsils), vasculitis, acute renal tubular necrosis, non-suppurative encephalitis,
and multifocal necrosis in all of the organs studied. The main differences with
other highly pathogenic strains (fowl plague (H7N7), Ck/Scot/59 (H5N1), Tern/S.A.
(H5N3), Ty/Ont/66 (H5N9), and Ck/Penn/83 (H5N2)) were: greater frequency of
vascular injury, nephropathogenicity, pancreatic islet cell necrosis, and diffuse
necrosis of lymphoid tissue. These findings collectively constitute a distinctive
feature of the strain under study. Influenza virus was detected in lung tissue
by RT-PCR, regardless of the viral dose, as well as in encephalon and kidney
in seven of eight doses used. These results are indicative of viral replication
in the three organs studied; hence, the pathologic changes can be attributed
to viral replication, without excluding other mechanisms of cell injury.
REFERENCES
Comité de Enfermedades Exóticas de la Asociación de Sanidad de los Estados Unidos. Enfermedades exóticas de los animales su prevención, diagnóstico y control. Comisión México - Americana para la Prevención de la Fiebre Aftosa. México (DF), 1986.
Easterday BC, Hinshaw VS. Influenza. In: Calnek BW, Barnes HJ, Beard CW, Reid WM, Yorder HW, editors. Diseases of Poultry. 9th ed. Ames (Io): Iowa State University Press, 1991:532-551.
Murphy B R, Webster RG. Orthomyxoviruses. In: Fields BN, Knipe DM, Howley PM, et al., editors. Fields Virology. Philadelphia: Lippincott – Raven Publishers, 1996:1397-1445.
Alexander DJ. A review of avian influenza in different bird species. Vet Microbiol 2000; 74: 3-13.
Fenner FJ, Gibbs EPJ, Murphy FA, Rott R, Studdenrt MJ, White DO. Veterinary virology. 2nd ed. New York: Academic Press Inc., 1993: 511-522.
Hooper PT. Lesions in chickens experimentally infected with 1985 H7N7 avian influenza virus. Aust Vet J 1989; 66(5):155-156.
Acland HM, Silverman BLA, Eckroade RJ. Lesions in broiler and layer in an outbreak of highly pathogenic avian influenza virus infection. Vet Pathol 1984;21:564-569.
Jungherr EL, Tyzzer EE, Brandly CA, Moses HE. The comparative pathology of fowl plague and Newcastle disease. Am J Vet Res 1946; 7: 250-288.
Van Campen H, Easterday BC, Hinshaw VS. Virulent avian influenza A virus: Their effect on avian lymphocytes and macrophages in vivo and in vitro. J Gen Virol 1989; 70: 2887-2895.
Mullis KB, Faloona F, Scharf SJ, Saiki RK, Horn GT, Erlich HA. Specific enzymatic amplification of DNA in vitro. The polymerase chain reaction. Cold Spring Harb Symp Quant Biol 1986; 51: 263-273.
Dispositivo Nacional de Emergencia de Sanidad Animal: Influenza aviar. Secretaría de Agricultura, Ganadería y Desarrollo Rural. Boletín No. 10. Dirección General de Sanidad Animal. 1995: 36.
Mohanty SB, Dutta SK. Virología Veterinaria. México (DF): Interamericana. 1983.
Luna LG. Manual of histologic staining methods of the Armed Forces Institute of Pathology. New York: Mc-Graw-Hill, 1968.
Gruber AD, Greiser-Wilke IM, Haas L, Hewicker-Trauwein M, Moennig V. Detection of bovine viral diarrhea virus RNA in formalin-fixed, paraffinembedded brain tissue by nested polymerase chain reaction. J Virol Methods 1993; 43: 309-320.
Shibata D. Identification of mismatched fixed specimens with a commercially available kit based on the polymerase chain reaction. Anat Pathol 1993;100 (6): 666-670.
Kwok S, Kiguchi R. Avoiding false positives with PCR. Nature 1989; 339: 237-238.
Ausubel FM, editor. Current protocols in molecular biology. New York: John Wiley & Sons, Inc. 1994.
Sambrook J, Fritsch EF, Maniatis T. Molecular cloning. A laboratory manual. 2nd. ed. New York: Cold Spring Harbor Laboratory Press, 1989.
Mies C. A simple, rapid method for isolation RNA from paraffin-embedded tissues for reverse transcription-polymerase chain reaction (RT-PCR). J Histochem Cytochem 1994; 42 (6): 811-813.
Riddell C. Developmental, metabolic, and miscellaneous disorders. In: Calnek BW, Barnes HJ, Beard CW, Reid WM, Yorder HW, editors. Diseases of Poultry. 9th ed. Ames (Io): Iowa State University Press, 1991: 827-862.
Riddell C: Avian histopathology. Lawrence, Kansas: Allen Press Inc. 1987.
Van Campen H, Easterday BC, Hinshaw VS. Destruction of lymphocytes by a virulent avian influenza A virus. J Gen Virol 1989; 70: 467-472.
Hinshaw VS, Olsen CW, Dybdahls SN, Evans D. Apoptosis: a mechanism of cell killing by influenza A and B viruses. J Virol 1994; 68 (6): 3667-3673.
Brown CC, Olander HJ, Senne DA. A pathogenesis study of highly pathogenic avian influenza virus H5N2 in chickens, using immunohistochemistry. J Comp Pathol 1992; 107: 341-348.
Hooper PT, Russell GW, Selleck PW, Stanislawek WL. Observations on the relationship in chickens between the virulence of some avian influenza viruses and their pathogenicity for various organs. Avian Dis
Shalaby AA, Slemons DR, Swayne ED. Pathological studies of A/Ckickens/Alabama/7395/75 (H4N8) influenza virus in specific-pathogen-free laying hens. Avian Dis 1994; 38: 22-32.
Slemons RD, Swayne DE. Replication of a waterfowlorigin influenza virus in the kidney and intestine of chickens. Avian Dis 1990; 34: 277-284.
Swayne DE, Slemons RD. Renal pathology in specificpathogen-free chickens inoculated with a waterfowlorigin type A influenza virus. Avian Dis 1990; 34:285-294.
Swayne DE, Radin MJ, Hoepf TM, Slemons RD. Acute renal failure as the cause of death in chickens following intravenous inoculation with avian influenza virus A/Chickens/Alabama/7395/75 (H4N8). Avian Dis 1994; 38: 151-157.
Cotran RS, Kumar V, Robbins SL. Patología estructural y funcional. 4ta ed. México (DF): Interamericana/McGraw-Hill, 1990.
Taubenberger JK, Reid AH, Krafft AE, Bijwaard KE, Fanning TG. Initial genetic characterization of the 1918 “Spanish” influenza virus. Science 1997; 275:1793-1796.
Schwarz TF, Zaki SR, Morzunov S, Peters CJ, Nichol ST. Detection and sequence confirmation of sin nombre virus RNA in paraffin-embedded human tissues using one-step RT-PCR. J Virol Methods 1995; 51: 349-356.