Tissue drug residue research often involves the killing of an animal every time tissue concentrations are determined. To decrease the number of animals required to perform tissue depletion studies and to circumvent the statistical problems associated with determining tissue depletion kinetic properties, using multiple animals, the renal depletion profile of gentamicin from individual sheep was studied, using a bilateral renal translocation technique. Seven ewes were surgically altered, allowed to stabilize, and then allocated into 2 groups; groups-1 sheep (n = 4) were given 3 mg of gentamicin/kg, IM, q 12 h for 10 days, and group-2 sheep (n = 3) were not given gentamicin. The kidneys from all ewes were biopsied 9 times over 74 days after the termination of gentamicin treatment. The renal concentrations of gentamicin were measured by use of a validated tissue digestion procedure coupled with a liquid-phase fluorescence polarization immunoassay. On days 75 and 77 after the end of gentamicin treatment, all ewes were euthanatized and necropsied. The concentrations of gentamicin in the biopsy specimens ranged from 71.9 to 183 microgram/g on days 1 and 2 after dosing, and decreased to concentrations ranging from 3.99 to 7.35 microgram/g on days 73 and 74 after the end of dosing. The decrease in renal gentamicin concentrations was best described by a biexponential equation, The early phase half-life was 2.8 days, whereas the terminal phase half-life was 59 days (harmonic means). There was no difference in the appearance or histologic features of the kidneys from groups 1 and 2. The only lesions noticed were linear fibroses that were attributed to the biopsy procedure.

A study was undertaken to determine the pressor and toxic effects of etoposide, an antineoplastic agent, when administered IV in 0.9% sodium chloride solution (0.4 mg of etoposide/ml) over a 30-minute period to dogs at a dosage of 40 mg/m2 of body surface. On day 1, 6 adult German Shorthaired Pointers were anesthetized with halothane, and blood pressures were measured via a femoral artery catheter before, during, and after the etoposide was administered. Systolic, diastolic, and mean blood pressures of each dog increased significantly (P less than 0.01) within 30 minutes after initiation of etoposide infusion. On day 3, when the dogs were not anesthetized, etoposide was again administered to each dog, using the same dosage. Each dog developed a moderate to severe cutaneous reaction characterized by moderate to severe pruritus, urticaria, and swelling of the head and extremities that began during the second infusion of etoposide. These same cutaneous reactions were seen on day 30, when etoposide was administered to 3 of the previously treated dogs and 2 previously untreated Beagles. We concluded that the administration of the commercial preparation of etoposide is likely to cause a significant reduction in blood pressure of anesthetized dogs, and that the drug is likely to induce a moderate to severe cutaneous reaction when administered to unanesthetized dogs.

Holstein cow 1 was examined because of skin fragility and delayed healing of skin wounds, which were markedly exacerbated around the time of parturition. A skin biopsy sample was obtained, and light microscopy revealed irregular deposition of thin collagen fibers in a dermal matrix. Although diffuse inflammation did not occur, the number of plump fibroblasts was increased. Electron microscopy revealed poor construction of collagen fibrils in the dermal matrix. Biochemical analysis of the dermis revealed a normal amount of collagen and uronic acid, but sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveled an increased proportion of soluble alpha-, beta-, and gamma-collagen chains of normal molecular weights. Neither procollagen nor its intermediates devoid of amino- or carboxy-terminal extension peptide were observed. Dermal collagen from cow 1 was more soluble in a neutral salt solvent, 0.5M acetic acid, and the acid containing pepsin than was dermal collagen from healthy cow 2. The peptic digestion profile of dermis from cow 1 revealed a lowered degree of intermolecular cross-linking and destabilization of helical structure in the dermis collagen. The extrahelical peptic cleavage of collagen before cyanogen bromide digestion resulted in release of more fragments derived from carboxy-terminal part of alpha1 chains in dermis of cow 1 than in dermis of healthy cow 2.

A bovine herpesvirus-1 (BHV-1) isolate (FI) from an aborted fetus was used to infect 9 heifers at various stages of gestation. Two heifers were inoculated IV on postbreeding day (PBD) 1, 7, or 14, and 3 heifers were inoculated in the sixth month of pregnancy. Plasma progesterone assays were used to monitor corpus luteum function in heifers inoculated during early pregnancy. Low progesterone values and infertility were seen in the 2 heifers inoculated on PBD 1. Luteal function remained normal in heifers inoculated on PBD 7 or 14. These 4 heifers inoculated on PBD 7 or 14 carried their fetuses to term, and their calves were free of BHV-1 infection at birth. Three heifers inoculated during the sixth month of pregnancy also carried their fetuses to term. Two calves were born alive, and BHV-1 was not isolated from nasal swab samples of either calf; the third calf was stillborn. Virus was not isolated from the stillborn calf's tissues, but BHV-1 was isolated from the placenta. Lesions were not detected in several tissues examined by light microscopy, and BHV-1 antigen was not detected by immunohistochemical examination of paraffin sections. Restriction endonuclease analysis of viral DNA was used to compare the FI virus to other BHV-1 isolates (Colorado-1, Iowa, and K22). On the basis of restriction endonuclease analysis, the FI isolate should be classified as a type-2 (infectious pustular vulvovaginitis) virus, specifically subtype a.

A study was conducted to examine sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood, including variation attributable to isolation of neutrophils from blood, variation between duplicate determinations of percentage phagocytosis, and the variation in the ability of neutrophils isolated from blood (over repeated collections from the jugular vein) to phagocytose. For the phagocytosis assay, jugular venous blood from each of 4 cows was divided into 2 equal portions. The neutrophils were isolated by lysis of red blood cells with 0.2% sodium chloride. The neutrophils (2 X 10(7)) were incubated in duplicate with 32P-labeled Staphylococcus aureus ([32P]SA; 2 X 10(8)) inskimmed milk samples (2.5% final concentration) prepared from 4 cows. This process was repeated thrice on neutrophils isolated from 4 cows at 2-week intervals. The proportions of variation in percentage of 32P-labeled S aureus phagocytosed between duplicate neutrophil isolations and between duplicate assay determinations were 0 and 1%. Differences among skimmed milk sources and among runs, using blood neutrophils taken at different times from the same donor cow, accounted for 62 and 36% of the total variation. The results indicated that variation arising from blood neutrophil isolation introduced into a phagocytosis assay within a single-day trial is of no concern. The large variation among skimmed milk sample sources indicated differences among cows in the ability of their milk to support phagocytosis. The variation in neutrophil isolations over time for any cow was considered too large to allow for evaluation of physiologic and environmental effects on phagocytosis of neutrophils isolated from blood.

An indirect immunoperoxidase staining technique was evaluated for detection of Mycoplasma hyopneumoniae in formalin-fixed, paraffin-embedded porcine lung. Lungs from swine with induced (n = 4) or naturally occurring M hyopneumoniae infection (n = 31) were examined grossly, by light and immunofluorescent microscopy, and by an indirect immunoperoxidase test, using antibody raised in swine against M hyopneumoniae as the primary antibody. Organisms stained by the indirect immunoperoxidase method were identified in tissue sections as pleomorphic brown-staining structures corresponding to those observed with immunofluorescence. Mycoplasma hyosynoviae, M hyorhinis, and Acholeplasma laidlawii did not stain with the indirect immunoperoxidase method, using antibody raised against M hyopneumoniae.

Tracheas from 15 toy breed dogs with normal tracheas and 6 dogs with collapsed tracheas were examined histologically and histochemically. Tracheal cartilage was analyzed for chondroitin sulfate by means of alcian blue (CEC method) and for calcium with alizarin red S. Cartilage arcs from dogs with collapsed tracheas had areas that were apparently hypocellular, and some had other areas that appeared like fibrocartilage or fibrous tissue. Histochemically, collapsed tracheal cartilage had less chondroitin sulfate and calcium than did normal tracheal cartilage. Cartilage arcs from the collapsed tracheas were not uniformly affected to the same degree, and parts of a given tracheal arc appeared normal, whereas other parts had an abnormal histologic appearance.

To study communications and boundaries of the middle carpal and carpometacarpal joints of the horse, 50 forelimbs were obtained from fresh cadaver specimens. Blue latex solution (20 +/- 2.5 ml) was injected into the middle carpal joint, and the specimens were frozen in extension. Frozen specimens were cut into 1-cm sagittal sections from the middle of the radius to the middle of the metacarpus. The communications between the middle carpal and carpometacarpal joints and the presence, length, and position of the distopalmar outpouchings of the carpometacarpal joint were recorded. The middle carpal and carpometacarpal joints always communicated between os carpale III (C3) and os carpale IV (C4). An additional communication between the joints existed in 17 (34%) of the specimens, 10 on the palmar aspect of C4, and 3 on the palmar aspect of os carpale II (C2). When os carpale 1 (C1) was present (n = 5), communication between C1 and C2 was observed in 4 of the 5 specimens. In all specimens, medial and lateral distopalmar outpouchings of the carpometacarpal joint were observed and were located between the axial surface of os metacarpale II (MC2) and os metacarpale IV (MC4) and the abaxial surface of the suspensory ligament. There was no significant difference between the lengths of the lateral (2.3 +/- 0.54 cm) or medial (2.6 +/- 0.75 cm) distopalmar outpouchings. Small extensions from the distopalmar outpouchings were seen and extended axially into the fibers of the suspensory ligament or between the suspensory ligament and the distal accessory ligament of the deep digital flexor tendon. In one carpus, the middle carpal joint communicated with the antebrachiocarpal joint between the articulation of the os carpi intermedium (Ci) and the os carpi ulnare (Cu).

In an attempt to isolate monoclonal antibodies specific for bovine lymphocytes, spleen cells from mice immunized with bovine lymphocytes were fused with the mouse myeloma cell line SP-2/0. The resulting hybridoma cell lines were tested for reactivity with bovine lymphocytes, polymorphonuclear neutrophils, RBC, gamma-globulin, kappa-casein, beta-casein, alpha-S1-casein, and beta2-microglobulin (beta2m) and with beta2m from rabbits, goats, and human beings. None of the clones secreted anti-bovine lymphocyte-specific antibody. However, 4 secreted monoclonal antibodies to bovine beta2m. They also reacted with beta2m from rabbit, goat, and human being. One monoclonal antibody also was found to be reactive with bovine immunoglobulin. Monoclonal antibodies to beta2m could serve as a tool to (1) exlore the homology of the beta2m molecule among various species, (2) examine the relationship of beta2m with the constant region of the immunoglobulin molecule, (3) quantitate bovine beta2m in various body fluids and major histocompatibility antigens on cell surfaces, (4) help characterize those antigens in cattle, and (5) be used for tissue typing of those antigens.

Adult Beagles were used to evaluate clinical signs and serologic response after inoculation with, or exposure to, Borrelia burgdorferi. An indirect immunofluorescent assay (IFA) and 2 ELISA were used to monitor the serologic response to B burgdorferi. Feeding infected ticks on 4 dogs (group 1) failed to cause seroconversion, and SC inoculation with 500 organisms caused minimal seroconversion in 2 of 4 dogs (group 2). At 56 days, approximately 3.01 X 10(8) B burgdorferi organisms were injected IV into group-1 dogs, and intraperitoneally into group-2 dogs. A control group of 4 dogs (group 3) had noninfected ticks feed on them, and then were given IV injection of physiologic saline solution. Increases in immunoglobulin M (IgM) titers were detected in 2 of 4 group-2 dogs approximately 7 days after the initial exposure. These titers returned to negligible values 20 days later. Immunoglobulin G titers increased approximately 10 days after the initial exposure and were mildly increased 56 days later, when dogs were exposed a second time. Both the IV and intraperitoneal injections (second exposures) resulted in increased IgM titers, which in both groups eventually returned to preexposure values after approximately 2 months. Immunoglobulin G titers increased within a week after the second exposure, and in 3 dogs monitored for 8 months, returned to negligible values after the 8-month period. One control dog had a slightly increased IgG titer 24 days after the second inoculation. The possibility of urine transmission is suggested. Clinical status, hemograms, serum biochemical profiles, ECG and results of urinalyses remained normal throughout the study. Borrelia burgdorferi was not isolated from either the blood or urine of these dogs. Gross or microscopic pathologic changes were not detected on necropsy.