1975;11:92\94. and viral DNA were tested by counter\immunoelectrophoresis (CIEP) and PCR, respectively. Animals that were PCR or CIEP positive at 196 dpi (n?=?41) were killed at 218 dpi, and samples of blood and seven organs were tested by CIEP and PCR. Results Antibody production persisted in all seroconverted mink until the termination of the experiment, whereas 71.1% of the mink showed short\lived viremia. Significant associations were observed between inoculum dose and the incidence of viremia until 84 dpi which disappeared thereafter, whereas associations between inoculum dose and the incidence Rabbit Polyclonal to Gastrin of seropositive mink were significant on all sampling occasions. Antibody titer at 218 dpi significantly decreased with decreasing inoculum dose. AMDV DNA was detected Cetirizine Dihydrochloride in the bone marrow, lymph nodes, and spleen samples of almost all mink inoculated at every dose but was not detected in other organs of some mink. Conclusions CIEP is usually more accurate than PCR for detecting AMDV contamination in mink. Using antibody titer in naturally infected mink may not be accurate for the identification of tolerant mink. values in this analysis do not measure the strength of agreements, rather they test whether the estimated kappa coefficient is not due to chance. Associations among the incidence of sprinklers and antibody titer were calculated by Spearman’s rank correlation. The Probit process with the logistic distribution was used to calculate 50% infectious dose (ID50) of the 10% spleen homogenate using CIEP and PCR results at 20, 35, and 56 dpi. 3.?RESULTS 3.1. Viral DNA in plasma Six inoculated mink died during the course of the study, one from each of the 100, 10?4, 10?6, and 10?7 doses and two from 10?3 dose (Table?1). Four of the lifeless mink were PCR and CIEP positive from 20 or 35 dpi until death and showed minor (score 1) to severe (score 3) AD lesions on their kidneys and/or livers (Table?1). One mink in each of the 10?4 and 10?6 doses remained CIEP and PCR negative until death and did not show any AD lesions at necropsy. Table 1 Distribution of lifeless mink by inoculum dose, PCR, and CIEP results and sampling days, and AD lesion scores at necropsy thead th rowspan=”2″ style=”border-bottom:solid 1px #000000″ colspan=”1″ Dose /th th colspan=”5″ style=”border-bottom:solid 1px #000000″ rowspan=”1″ Days postinoculation a /th th colspan=”2″ style=”border-bottom:solid 1px #000000″ rowspan=”1″ AD lesion scores b /th th rowspan=”1″ colspan=”1″ 20 /th th rowspan=”1″ colspan=”1″ 35 /th th rowspan=”1″ colspan=”1″ 56 /th th rowspan=”1″ colspan=”1″ 84 /th th rowspan=”1″ colspan=”1″ 140 /th th rowspan=”1″ colspan=”1″ Kidneys /th th rowspan=”1″ colspan=”1″ Liver /th /thead 0+/++/++/+..12?3+/++/+…01?3+/C+/++/++/+.23?4C/CC/CC/CC/C.00?6C/CC/CC/CC/CC/C00?7+/C+/++/++/+.21ControlC/CC/CC/CC/C.00 Open in a separate window Abbreviations: AD, Aleutian disease; CIEP, counter\immunoelectrophoresis; PCR, polymerase chain reaction. aPCR result/CIEP result, + positive, C unfavorable, . animal was Cetirizine Dihydrochloride lifeless. b0, 1, 2, 3 are none, minor, moderate, and severe lesions, respectively. All mink inoculated with the 100 or 10?1 doses were PCR positive by 20 dpi, whereas some mink inoculated with the other doses remained PCR unfavorable until 196 dpi (Physique?1). Seven mink became PCR positive for the first time at 196 dpi and six mink (four juveniles and two adults) remained PCR unfavorable until 196 dpi; one in each of Cetirizine Dihydrochloride the 10?2, 10?5, 10?6, 10?7, and two in the 10?4 doses. Four of the six PCR unfavorable mink at 196 dpi, which were inoculated with 10?5 or higher doses and were kept until 218 dpi, became PCR positive at that time. Eight mink (five juveniles and three adults) and Cetirizine Dihydrochloride three juveniles, which became viremic at 20 or 35 dpi, respectively, remained viremic until 196 dpi, whereas viremia in 32 of the 45 mink (71.1%) that became PCR positive before and survived until 196 dpi, was Cetirizine Dihydrochloride irregular or short\lived. Logistic regression analyses revealed that the effects of viral dose on the incidence of viremia were significant until 84 dpi, but not at the subsequent sampling occasions. For each unit switch in dose (10 occasions dilution), the expected switch in log of odds for viremia at 20 dpi, adjusted for the age of mink, was 0.88, and gradually decreased at subsequent sampling occasions (Table?2). The estimate of odds ratio showed that the odds of viremia changed by 2.41 times for each unit change of inoculum dose at 20 dpi, and the estimates steadily decreased to 1 1.043 at 196 dpi. The predicted probabilities for viremia across all inoculum doses were the greatest at 20 dpi and the effect became smaller as occasions after inoculation prolonged, and almost disappeared after 84 dpi (Physique?2). The effects of age around the incidence of viremia were significant only at 20 dpi. The log of odds for viremia was lower in the juvenile than in the adult mink at 20 dpi (Wald em /em 2 (1)?=?4.78,.