The objective of this study was to evaluate the efficacy of regional intravenous (i.v.) injection of ceftiofur in delivery of this drug to joint fluid and plasma in a limb distal to a tourniquet in five, healthy, adult, mixed breed beef cattle. A tourniquet was positioned in the mid-metacarpal region, and 500 mg of ceftiofur was administered through a catheter in the dorsal common digital vein (DCDV). Plasma samples were collected from the catheter at 15, 30 and 45 min postinjection, and from the abaxial proper palmar vein (APPV) at 15 min postinjection. Synovial fluid was collected from the metacarpal phalangeal joint at 45 min postinjection. Ceftiofur concentrations were estimated in plasma and synovial fluid using high-pressure liquid chromatography (HPLC) and a microbiological assay utilizing Pasteurella haemolytica as the test organism. Both assays indicated highest plasma concentrations of ceftiofur at 15 min, with the concentrations declining with time. Concentrations of ceftiofur in plasma obtained from the DCDV were not significantly different from APPV levels, indicating rapid distribution of ceftiofur within the limb. Microbiological assay always demonstrated higher concentrations of ceftiofur compared with HPLC assay, because the former probably also detected the active metabolites of ceftiofur as well as the parent compound. At 45 min, ceftiofur concentrations determined by HPLC were 251+/-97 and 15+/-5 microg/mL in plasma and synovial fluid, respectively. Regional intravenous injection appears to be a feasible technique to produce rapid distribution of ceftiofur within the limb well above therapeutic concentrations.

We characterized an abundant late 1.7-kb cytoplasmic polyadenylated RNA (L1.7 RNA) transcribed from the bovine herpesvirus 4 (BHV-4) HindIII W fragment, in a region of the genome not conserved with Epstein-Barr virus and herpesvirus saimiri. L1.7 RNA contains only extremely short (<100 nucleotides) open reading frames followed by two repeat arrays. The first repeat array contains 11 copies of a 23-bp unit, TGGCACTA GTAGCATTTAACCCC. The second and third copies are each interrupted by 15-to 17-bp sequences that are identical to each other for the first 15 bp. In addition, the second and third copies of the repeat unit each contain two copies of nucleotides 5 to 9 (ACTAG) of the repeat unit, one at each end of the interruption. The second repeat array contains 12 copies of a 25-bp sequence, GCTGTGTATTATTGAGTATTTTTTA. The promoter-regulatory region of L1.7 was activated by the BHV-4 immediate-early gene 2 product (IE2), a homolog of the Epstein-Barr virus R transactivator, in cotransfection assays. We mapped an IE2 recognition site within a 167-bp fragment approximately 10 bp 5 to the start of L1.7 RNA transcription, using cotransfection assays and gel retardation assays. Using gel retardation assays, we mapped an IE2-binding site within this fragment to a 31-bp region from 56 to 86 bp 5 to the start of L1.7 RNA transcription. This IE2-binding site was able to transfer IE2 responsiveness to a heterologous promoter. However, IE2 responsiveness was affected by both position and orientation. Alignment of the L1.7 IE2-binding site sequence with sequences of two other BHV-4 IE2-binding sites resulted in a provisional IE2-binding site consensus sequence different from the Epstein-Barr virus R transactivator-binding site.