| Avian leukosis viruses (ALV) is a retroviruses.It exists generally and can cause 1% ~ 2% adult chicken induce visceral tumor or death. The infection viruses can be divided into A, B, C, D, E and J six subtypes. Common clinical subtypes are J and A. E subtypes have no pathogenic. In the past ten years, ALV on meat-type chicken, Egg type chicken and local strains caused great economic losses. Nearly two years, The chicken leukemia cases rise significantly disease in China and become a danger to one of the main epidemics heritage layers. In recent years, avian leukosis / hemangioma increased by a significant hazard to the disease as the major diseases of egg industry, and found in the clinical typical cases of fibrosarcoma. respectively, from the clinical Hyline brown layers and laboratory brown SPF chicken get two showed clinical cases of fibrosarcoma, were carried out virus isolation and identification of human pathogenic trial.1 Hyline brown layers of acute fibrosarcoma diagnosis and artificially created diseaseIn June 2010, a layers factory of Shan Dong raise 8,000 layers. To 240 day, the mortality rate about 25%. There is a sick chicken in cardiac apex, sternum, inferior a sarcoma regiment among them. The test is based on this, collecting material and isolating identificating and making artificial disease test.1.1 tumor virus isolation and identificationGetting cancer prefiltration, both the cancer of grinding filtration liquid vaccinated DF-1 cells .cultivating 5 ~ 7d, after the cowslip had grown normal-looking, continue to cultivate 5 ~ 7d, take cell culture with poultry leukemia virus detection ALV p27 antigen. The antigen positive cells separately in JE9, 11B154, H19 and ALV A/B gp85 on mice on the cowslip serum layer of cells, the results showed that they infected by ALV-J or ALV-A/B, but does not exist MDV and REV infection. 1.2 Histopathology of tumor tissue observedHistopathology showed tumor cells sarcoma are array messy and cells of different sizes1.3 Sequence analysis of virus isolateProviral ALV isolated amplified 2.2kb genomic DNA, including part of the LTR region and complete env gene fragments, PCR amplification products were cloned and sequenced using the DNAStar software for sequence analysis of gene products. gp85 gene encoding the sequence of amino acids and other reference strains of the comparisongp85 genes of the isolates, whose amino acid sequence had over 87.5% of identity to 10 reference strains of subgroup J. However, The homology to subgroups A,B, C, D, E was in the range of 37.6% - 40.1%. The phylogenic tree indicates that the isolates definitely fell into subgroup J.gp37 gene encoding the sequence of amino acids and other reference strains of the comparisonThis strain gp37 coding the sequence of amino acids and J subtypes in reference strains of the homology 92.7% ~ 94.9% range, homology is higher. The homology to subgroups A,B, C, D, E was in the range of 60.8% - 78.9%.1.4 Hyline brown layers tumer pathogenicity artificial testTumor mass, add 3 times the volume of PBS grinding, until the filter into the filtered homogenate, sick material liquid.Hyline brown different age layers artificial disease testing Diseased stock solution, by SC and IA were inoculated 1d, 7d, 14d, 180d, 240d of Hyline brown layers. The different injection mode are able to reproduce tumor, merely tumor first appeared different time, 1d to 14 d age can appear in about 10 d, after 180 d of age in about 20 d to appear, but the incidence and mortality tumor were 100%. And we find that, the abram brown chicken neck subcutaneous injection group will not only caused the occurrence of neck, subcutaneous tumor can also cause the thymus and spleen, even atrophy of liver or spleen sarcoma or hemangioma. Intraperitoneal injection group can observe, except in muscular stomach and superior mesenteric, see the obvious tumor nodules outside, some still can cause liver and spleen caruncle and hemangioma, but can not cause cervical hypodermic tumour.1 day Hyline brown layers neck tumor growth of SC1d Hyline brown layers tumor growth, after the commencement of the poison attack within a week without any change .In growth to 9d, most of the chicken skin in the neck, touch to the grain size of mass, rapid tumor growth in the next few days, 5d-6d can be achieved after more than 30 mm in diameter.1.5 Isolation, identification and morphological observation of artificial tumor tissue isolation and identification of artificial tumor tissue according to the original methods The results showed that the artificial tumor pathogenicity test was the apparent existence of ALV-J infection, there may ALV-A/B sub-group of infection, but there is no infection of MDV and REV. Tumor disease material, Giemsa staining and histopathological observation shows that the individual heterogeneity tumor cells, diverse in shape, large nuclei, stain, and some mitotic figures can be found obvious typical fibroblast sarcoma .2 Pathological and viralogical analysis of a fibrosarcoma case induced by avian leucosis virus of subgroup ASDAU09C3 is separated from Hyline brown layers of ALV-A subgroup strain.Embryo yolk inoculation 5d SPF 3000 TCID50, growth to 7 months of artificial killed.kidney and liver sacrificed to obtain the performance of large nodules typical focal cases.2.1 Virus isolation,identification and sequence analysis of SPF chicken fibrosarcomaIFA test results show that kidney and liver tumors exist ALV-A/B infection, but not in ALV-J, MDV, REV infection. Liver and kidneys with the original attack strains isolated envelope protein gp85 gene encoding the amino acid sequence homology was 99.5%.2.2 SPF chicken fibrosarcoma histopathological observationSDAU09C3 strain caused by the tumor, mainly small collagen fibers, irregularly arranged cells 2.3 SPF chicken fibrosarcoma pathogenicity test of artificialLiver and kidney tumors grinding fluid filtration, attack 1d SPF chickens, the continuous observation to 75 days, liver, kidney tumors not detected in the leachate can be displayed on the biology of acute virus induced tumors.
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