Gross pathology, histopathology and immunohistochemistry of chickens experimentally inoculated with Km_5666.

<p>(A) Grayish-white ill-defined tissue was located in the septum at 35 days old. (B) Locally extensive growth of hypertrophied cardiomyocytes. Bar  = 75 µm. (C) Mitosis (arrowhead) observed in hypertrophied cardiomyocytes. Bar  = 30 µm. (D) Numerous nuclei in atypical myocardium were positive for PCNA. Bar  = 20 µm. (E–F) Hypertrophied cardiomyocytes positive for p-Akt (E) and p-tuberin (F). Bars  = 20 µm.</p

The degree of cardiac lesions in chickens experimentally infected with Km_5666 and Km_5892.

a<p>The degree of hypertrophied cardiomyocytes with atypical nuclei was defined as follows: +, individual cells; ++, multiple and/or small clusters; +++, multiple and extensive growth.</p>b<p>The degree of non-suppurative myocarditis was defined as follows: +, focal or multifocal (under 25 µm in diameter); ++, multifocal (from 25 µm to 200 µm); +++, multifocal (over 200 µm).</p>c<p>The degree of fibrosis was defined as follows: +, interstitial; ++, multifocal interstitial (under 400 µm in diameter); +++, multifocal interstitial (over 400 µm in diameter).</p>d<p>Total number in 10 high-power field.</p

Phylogenic tree based on <i>pol</i> gene of the isolates.

<p>Phylogenic tree constructed by the neighbor-joining method showing the relationships among Km_5892, FGV prototype and standard ALSVs based on the <i>pol</i> gene. The nucleotide sequences were aligned with the Clustal W 1.8.3 program. Bootstrap values of 1000 trials using the neighbor-joining method. Scale bar corresponds to a distance of 0.03 as the frequency of amino acid substitutions in the pairwise comparison of two sequences according to the Kimura two-parameter method.</p

Sequence of the oligonucleotide primers and targets.

<p>Sequence of the oligonucleotide primers and targets.</p

Histopathology and immunohistochemistry of hearts in Japanese native fowls.

<p>(A) Mononuclear cells infiltrate into myocardial fibers. Chicken No. 5. Bar  = 40 µm. (B) Disarrangement of myocardial fibers with interstitial fibrosis. Chicken No. 9. Bar  = 90 µm. (C) Interstitial myocardial fibrosis confirmed by Masson's trichrome stain. Chicken No. 14. Bar  = 25 µm. (D) Hypertrophied cardiomyocytes with multiple atypical nuclei. Mitosis (arrowhead) is occasionally noted in these cardiomyocytes. Chicken No. 14. Bar  = 20 µm. (E) Matrix inclusion bodies (arrowhead) in the cytoplasm of Purkinje fibers. Chicken No. 14. Bar  = 20 µm. (F) Purkinje fibers and matrix inclusion bodies positive for ALV common antigen. Chicken No. 14. Bar  = 20 µm. (G) A large atypical nucleus in a hypertrophied cardiomyocyte is positive for PCNA. Chicken No. 14. Bar  = 20 µm.</p

Clinical signs, gross findings, cardiac histopathology and results of ALV isolation in chickens.

a<p>M: male, F: female</p>b<p>The degree of hypertrophied cardiomyocytes with atypical nuclei was defined as follows: +, individual cells; ++, multiple and/or small clusters; +++, multiple and extensive growth.</p>c<p>The degree of non-suppurative myocarditis was defined as follows: +, focal or multifocal (under 25 µm in diameter); ++, multifocal (from 25 µm to 200 µm); +++, multifocal (over 200 µm).</p>d<p>The degree of fibrosis was defined as follows: +, interstitial; ++, multifocal interstitial (under 400 µm in diameter); +++, multifocal interstitial (over 400 µm in diameter).</p>e<p>Total number in 10 high-power fields.</p>f<p>+: positive,−: negative,ND: no data.</p