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Infection with avian influenza virus (AIV) in laying birds field cases often lead to clinical signs of torticollis and curled toe paralysis. Clinical signs of torticollis and curled toe paralysis is similar to the clinical signs of Newcastle disease virus infection in poultry, making it difficult for the confirmation of diagnosis of infection with both viruses. Immunopathological immunohistochemistry streptavidin-biotin (IHC SB) is an antibody-based test to detect the presence of pathogens, especially AIV in poultry with the principle of antigen detection in tissue specimens. This research aims to detect AIV infection in laying birds field case with clinical signs of torticollis and curled toe paralysis. The samples (brains from 20 layer chicken) were taken from the cases of the disease in poultry in several commercial poultry farms. The chickens showed clinical signs of torticollis and curled toe paralysis, and oedema in the brain suspected of being infected AIV. After being necropsied, then the brains were tested IHC SB and observed with a digital microscope camera system and analyzed descriptively qualitative. The results showed that the avian influenza virus antigens was found in the parenchyma, neural and endothelial cells of brain blood vessels. Based on these results, we can conclude that Immunopathologic immunohistochemical staining of streptavidin-biotin can be applied for confirmation of the diagnosis of avian influenza virus (AIV) on the brain of commercial laying chickens showing torticollis and curled toe paralysis. Keywords : AIV; IHC SB; brain; torticollis; curled toe paralysis

Poultry is one of animal protein sources that have been consumed much in Indonesia. Various of health disorders are caused by environmental changes, woof and infectious agents have caused decline in production and economic loss. One of diseases that have a big impact on poultry husbandry is Newcastle disease (ND). Newcastle disease virus (NDV) is an important disease in Indonesia, because this has spread around Indonesia and caused a big loss for poultry industry moreover, this has high morbidity and mortality and very rapid spread of the virus. The purpose of the research is to find out and determine a fast, exact and accurate, and efficient and effective application immunohistochemistry streptavidin biotin (IHK SB) test on NDV diagnosis affirmation and for NDV fast detection as a cause of disease on commercial laying hens with clinical torticollis and curled toe paralysis symptoms. The samples used in the form of tissues from 20 commercial laying hens with an indication of clinical torticollis and curled toe paralysis symptoms. Brains would be tested using streptavidin biotin. Inspection result data obtained by immunohistochemistry streptavidin biotin (IHK SB) method is analyzed in a descriptive and qualitative way. The result of the research proves that IHK SB coloration, NDV antigen could be detected on brains. Based on the result of the research, it could be concluded that IHK SB could be applied for NDV diagnosis affirmation and it proves that commercial laying hens with clinical torticollis and curled toe paralysis symptoms. Keywords: Newcastle disease virus, immunohistochemistry streptavidin biotin, torticollis, curled toe paralysis

THE early recognition of the importance of milk products as riboflavin sources in poultry rations has led to numerous riboflavin assays of these products, based largely on chick growth. Heiman (1935) showed the riboflavin content of dried skimmilk and of dried whey to be in a ratio of 1:1.5. Norris et al. (1936) confirmed this result and established the riboflavin content of dried whey and dried skimmilk at 30 and 20 micrograms per gram, respectively. Heiman and Carver (1937), assaying samples of neutralized dried whey, dried skimmilk, and dried sweet-cream buttermilk, obtained from the same processing plant and prepared within one week in April, concluded that the whey contained between 10 and 20 percent more riboflavin than the dried skimmilk and that the dried sweet-cream buttermilk contained 90 to 100 percent more riboflavin than the dried skimmilk and 60 to 70 percent more than the neutralized dried whey. Jukes (1937) . . .

LEPKOVSKY and Jukes in 1936 showed that riboflavin was essential in chick nutrition. Similar observations were noted by Bethke, Record, and Wilder (1937). The later investigators also reported that riboflavin was effective in preventing the curled-toe paralysis originally described by Norris, Heuser, and Wilgus (1930). Stokstad and Manning (1938) observed that large amounts of riboflavin completely prevented curled-toe paralysis whereas small additions to the basal ration permitted the condition to develop. These findings are in accord with the earlier report of Norris et al. (1936) that the addition of small quantities of vitamin G supplement to the basal ration often increased the incidence of curled-toe paralysis whereas larger amounts decreased it. This led Norris et al. (1936) to suggest that the curled-toe paralysis was due to a deficiency of some other factor rather than to a deficiency of the growth-promoting vitamin G. The problem was greatly clarified when Phillips and . . .