Bovine blood mononuclear cells were separated into 2 fractions by use of centrifugl elutriation. Total recovery, as well as recovery of each fraction, was greater than that obtained by use of Ficoll-sodium diatrizoate separation. The lymphocyte fraction contained less than 1% granulocytes, and the granulocyte fraction contained only 7% lymphocyte contamination. The technique was reproducible and results proved to be comparable with those of Ficoll-sodium diatrizoate density-gradient centrifugation; furthermore, the method is considerably cheaper and less time-consuming for processing large volumes of blood. Viability of cells separated by elutriation always was greater than 98%, whereas viability of cells separated by Ficoll-sodium diatrizoate was greater than 95%. Also, mitogen activation of lymphocytes separated by elutriation was superior to that of lymphocytes separated by Ficoll-sodium diatrizoate centrifugation.

To evaluate herd-level risk factors for seropositive status of cattle to 1 or more bluetongue viruses. 110 herds of cattle in Nebraska, North Dakota, and South Dakota. Blood samples were collected before and after the vector season. Samples were tested for antibodies against bluetongue virus by use of a commercially available competitive ELISA. Factors evaluated included descriptors of geographic location and management practices. Trapping of insect vectors was conducted to evaluate vector status on a subset of 57 operations. A multivariable logistic regression model was constructed to evaluate associations. For the full data set, altitude and latitude were associated with risk of having seropositive cattle (an increase in altitude was associated with an increase in risk, and a more northerly location was associated with a decrease in risk of a premise having seropositive cattle). Import of cattle from selected states was associated with an increase in risk of having seropositive cattle. From the subset of herds with data on vector trapping, altitude and latitude were associated with risk of having seropositive cattle, similar to that for the full model. However, commingling with cattle from other herds was associated with a decrease in risk of seropositivity. Findings reported here may be useful in generating additional hypotheses regarding the ecologic characteristics of bluetongue viruses and other vector-borne diseases of livestock. Sentinel surveillance programs are useful for documenting regionalization zones for diseases, which can be beneficial when securing international markets for animals and animal products.