All adult mice injected with the te

All adult mice injected with the test article or the negative control article appeared normal and healthy for the twenty-eight day observation period.All suckling mice injected with the test article or the negative control article appeared normal and healthy after 14 days.The surviving mice of each group were homogenized and the homogenate of each group was passaged into a new group of suckling mice.The remainder of the homogenates was frozen at -70°C ± 10°C.In the blind passage, all suckling mice injected with the test article homogenate or the negative control article homogenate appeared normal and healthy after 14 days.All of the test article and all of the negative control article injected guinea pigs remained normal and healthy during the twenty-eight day test period.See Tables 2 and 3 for a summary of the data discussed above.Ten nine day old embryonated hens' eggs were injected by the allantoic route with the test article or the negative control article.These eggs were examined for viability at 24 hours and three days post-injection and allantoic fluids from day 3 were tested for hemagglutination using chicken, human O, and guinea pig erythrocytes.All eggs were viable and fluids were negative for hemagglutination.The day 3 fluids from each group were pooled and injected into ten new nine day old eggs using the same route of injection.These eggs were examined for viability at 24 hours and three days post-injection.All of the eggs injected with the pooled allantoic material from the test article or negative control article injected eggs appeared viable at 24 hours post-injection.At examination on day 3, all of the test article subpassage eggs and nine of the ten negative control article subpassage eggs contained viable embryos.One of the negative control article subpassage eggs contained a non-viable embryo.No growth was observed on blood agar plates streaked with fluid from the non-viable negative control article subpassage egg.The cause of death of this embryo could not be determined.(See Criteria for a Valid Test/Evaluation of Test Results.)Allantoic fluids from all subpassage eggs were tested for hemagglutination using chicken, guinea pig, and human O erythrocytes.All fluids were negative for hemagglutination.See Tables 4 and 5 for a summary of the data.Ten seven day old embryonated hens' eggs were injected by the yolk sac route with the test article or the negative control article.All of the eggs injected with the test article or the negative control article appeared viable at 24 hours post-injection.At 48 hours, all of the test article injected eggs and nine of the negative control article injected eggs appeared viable.At examination on day 9, all of the test article injected eggs and nine of the ten negative control article injected eggs contained viable embryos.One of the negative control article injected eggs contained a non-viable embryo.Growth identified as Bacillus was observed on blood agar plates streaked with fluid from the non-viable negative control article injected egg.The death of this embryo likely resulted from the bacterial contamination.The yolk sac material from all viable eggs in each group was pooled.A 10% suspension of pooled yolk sac material was injected into ten new seven day old embryonated eggs using the same route of injection.All of the embryos were viable.No evidence of viral contamination was observed due to the test article, SB-MCB-ACC-454-100 12 Feb 96 31 Jul 2000.The test will be considered valid if ninety percent of the control adult mice, eighty percent of the control suckling mice, eighty percent of the control embryonated hen's eggs, and seventy-five percent of the control guinea pigs survive the observation period, show no lesions at the site of injection or signs of viral infection.There may be instances when the test article animals meet the evaluation criteria, but the negative control animals do not meet the criteria detailed above, yet the assay will be considered valid.