Thirty-eight tracheobronchial aspirates (TBA) were collected from twenty 1 to 6-month-old foals, which were free of clinical signs of respiratory tract or other infectious disease. We collected TBA from 9 of the foals 3 times when they were approximately 8, 16, and 24 weeks old. Aspirates were examined cytologically after staining with modified Wright-Giemsa, Gram, toluidine blue, and prussian blue stains. Aerobic bacterial culturing was performed on all aspirates. Of the 20 initial TBA, 4 (20%) were normal cytologically on the basis of previously defined criteria for TBA from clinically normal horses, 6 (30%) had a high percentage of eosinophils (> 5%), 8 (40%) were classified as indicative of subacute inflammation, and 2 (10%) were classified as indicative of acute inflammation. Nine (45%) were positive for mast cells and none were positive for hemosiderin-laden macrophages (hemosiderophages). Of the 9 foals from which samples were collected at 16 and 24 weeks of age, results were similar, except for an increase in the number of TBA classified as indicative of chronic inflammation (33% and 22% respectively) and the number positive for hemosiderophages (33% and 88%, respectively). One TBA was considered nondiagnostic because of pharyngeal contamination. Culturing of 12 of the 37 aspirates (32%) yielded a potential microbial pathogen. Only 2 were positive cultures from the same foal. The following organisms were isolated: beta-hemolytic Streptococci spp (4), Actinobacillus/Pasteurella spp (4), Rhodococcus equi (2), unidentified nonenteric Gram-negative rod (1), and Escherichia coli (1). Thirty-four of the 37 aspirates (92%) yielded light growth of various organisms considered to be nonpathogenic and normal inhabitants of the upper respiratory tract. It was concluded that the presence of inflammatory cells, eosinophils, and mast cells in the tracheobronchial aspirates from clinically normal foals is a common finding. These cytologic findings were consistent in the samples collected from foals at 8, 16, and 24 weeks of age. It was also concluded that bacteria with recognized pathogenicity can be isolated from TBA from clinically normal foals and were most frequently isolated from 1- to 2-month-old foals or those with cytologic evidence of inflammation, even in the absence of clinical signs of respiratory tract disease.

Bovine pulmonary artery cells in cell culture were exposed to lipopolysaccharide (LPS) purified from Pasteurella haemolytica serotype A1. This resulted in severe membrane damage, which caused a time- and dose-dependent release of lactate dehydrogenase that was first detected 4 hours after exposure and reached a maximal mean release of 67% after 24 hours of exposure to 1 microgram of LPS/ml. Mean release of 51chromium followed by a similar pattern and reached a maximum of 61% following 24 hours of exposure to 10 micrograms of LPS/ml. Morphologically, endothelial cells responded to LPS by marked cell membrane retraction, the formation of numerous cytoplasmic blebs, and ruffling of the cell membrane. Subsequently, the cells became round and detached. Cell detachment reached a mean of 95% following 8 hours of exposure to 1 microgram of LPS/ml. These studies demonstrated that P haemolytica LPS is capable of causing direct damage to bovine pulmonary arterial endothelial cells, which may be important in the pathogenesis of bovine pneumonic pasteurellosis.

Monoclonal antibodies recognizing the O-polysaccharide portion of Brucella abortus strain 2308 provided BALB/c mice with passive protection against challenge exposure with the homologous strain. Numbers of colony-forming organisms in the spleen were reduced by IgM and IgG monoclonal antibodies. Active immunization of mice, using B abortus 2308S lipopolysaccharide, resulted in production of IgM antibody at 14 days. Clearance of organisms in the actively immunized mice after challenge exposure at 14 days was nearly identical to that in passively immunized mice. Mice either passively or actively immunized were effectively protected from 0 to 28 days. Bacterial colonization of the spleen was observed to increase in both groups of mice at 56 days and indicated that humoral responses were effective in eliminating the organism in the early stages of infection, but other immune mechanisms were necessary for protection of mice in the later stage of infection with virulent strains of B abortus.

Serum from dogs with surgically induced renal impairment was incorporated into the medium for erythroid bone marrow cultures. A significant correlation was found between serum activities of erythropoietin and numbers of erythroid colony-forming units grown in culture. Serum creatinine concentrations had no correlation, and serum parathyroid hormone activities had a negative correlation with numbers of erythroid colony-forming units that was below the level of significance. Purified 1-84 parathyroid hormone added to bone marrow cultures was found to be stimulatory to erythroid colony-forming unit growth in higher concentrations, but decreased the number of burst-forming units. Unmeasured substances in the canine serum appeared to have a greater effect on the canine erythroid bone marrow cultures than did creatinine or parathyroid hormone values.

Cell proliferation kinetic values were established for the epidermis, hair follicle epithelium, and sebaceous glands of 10 Beagles and 4 Cocker Spaniels with healthy skin and coats. Values were established by intradermal pulse-labeling injections of [3H]thymidine, examination of cutaneous biopsied tissues, and autoradiography. The epidermal basal cell-labeling index was 1.41 +/- 0.46% for Beagles and 1.71 +/- 0.56% for Cocker Spaniels. The hair follicle basal cell-labeling index was 1.46 +/- 0.78 and 1.07 +/- 0.42%, respectively. Calculated epidermal cell-renewal time for viable layers of the epidermis was 23.38 +/- 5.93 days for Beagels and 20.97 +/- 4.92 days for Cocker Spaniels. Differences between cell kinetics data for the 2 breeds were not significant (P greater than 0.05). The basal cell-labeling index for the sebaceous gland was significantly (P = 0.009) lower for Cocker Spaniels (0.40 +/- 0.18%) than for Beagles (1.81 +/- 1.08%). Seemingly, epidermal and follicular cell proliferation kinetics in healthy dogs was similar between the 2 breeds, whereas sebaceous gland basal cells were less proliferative in healthy Cocker Spaniels than in healthy Beagles.

The use of cultured tissue has not yet become wide-spread in research involving equine disease, and this may be attributable in part to the scarcity of published reports concerning tissue culture methods for this species. We report here the isolation of equine microvascular endothelium (EMVE) from fresh omental tissue of horses and ponies. Fresh donor tissue was minced, subjected to collagenase digestion, and filtered. Cells were layered on 5% bovine serum albumin for gravity sedimentation, the bottom layer was collected, and the cells were plated onto fibronectin-coated flasks. Medium consisted of Dulbecco modified Eagle medium with 10% whole fetal bovine serum (wFBS) and 20 micrograms of endothelial cell growth supplement/ml. The EMVE grew readily in culture, had the cobble-stone morphologic feature at confluence, stained positively for factor VIII-related antigen, and metabolized acetylated low-density lipoprotein. Fibroblast and smooth muscle cell contamination was minimal in primary cell cultures, which were successfully passed and maintained in culture for 3 to 5 serial passages, using various media and substrates. Preliminary studies were undertaken to determine optimal growth conditions with a range of variables: serum concentration, extracellular matrix components, and growth factors. Optimal conditions were achieved with a minimum of 10% wFBS, and with either fibronectin or laminin as extracellular matrix substrates. The EMVE grew adequately in Dulbecco modified Eagle medium plus 10% wFBS, and the added growth factors or serum supplements did not appear necessary for growth of EMVE.

Epidermal growth factor was injected intracamerally into the anterior chamber of the right eye of 9 cats. The central portion of the cornea in 8 of the 9 cats that had been cryoinjured. Effect of epidermal growth factor on the repair of endothelial cells in cats was evaluated by endothelial specular microscopy. Endothelial cell density and corneal thickness were studied quantitatively, as a measure of endothelial cell function. The repair process also was evaluated qualitatively by studying morphologic changes, developing as a result of reendothelialization and return to normal function. Seemingly, differences between rate of healing of cryoinjured eyes injected with epidermal growth factor and that in nontreated eyes were not significant (P = 0.86). The endothelial repair process was characterized by enlargement and migration of adjacent noninjured cells.

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.

Properties of the attachment of Treponema hyodysenteriae to Henle intestinal epithelial (HIE 407) cells were examined. The frequency of attachment depended on the motility and viability of the spirochetes. Rabbit hyperimmune and swine convalescent antisera inhibited attachment. Treatment of HIE cells with neuraminidase had no effect on attachment; however, treatment of spirochetes with the enzyme decreased adherence significantly (P = 0.01). Attachment was inhibited by N-acetylneuraminic acid, D-glucuronic acid, and fetuin. Adherence was increased following coincubation with N-acetylglucosamine or yeast mannan. Surface antigens of T hyodysenteriae, isolated by chemical extraction, competitively inhibited adherence. Concentrated T hyodysenteriae culture supernatant fractions inhibited adherence, but concentrated phosphate buffered-saline washings of the spirochete and concentrated uninoculated media did not inhibit adherence. Sialic acid was detected in unwashed T hyodysenteriae and spent culture supernatant fractions in higher concentrations than from washed spirochetes and uninoculated media. It was concluded that the binding adhesins on T hyodysenteriae for cultured HIE cells may contain sialic acid residues.

The genetic and antigenic nature of feline cell-associated herpesvirus (FeCAHV) was characterized by use of DNA restriction endonuclease analysis, and direct and indirect fluorescent antibody (FA) techniques. Serologic responses of 6 conventionally reared cats with induced FeCAHV urinary tract infection were retrospectively evaluated, using an indirect FA test. The EcoRI, HindIII, and Pst I restriction endonuclease cleavage patterns of FeCAHV DNA were similar to those of bovid herpesvirus 4 (BHV-4; DN599 strain) DNA. Specific fluorescence was observed when FeCAHV-inoculated cell monolayers were reacted with fluorescein-conjugated BHV-4 (DN599 strain) antiserum. Conversely, specific fluorescence was also observed when feline anti-FeCAHV serum and fluorescein-conjugated caprine anti-feline IgG was reacted with BHV-4 (DN599 strain)-infected cell monolayers. At postinoculation week 10, serum antibody titer in cats with FeCAHV-induced urinary tract infection ranged from 1:2,560 to 1:10,240, as measured by use of indirect FA testing. It was concluded that FeCAHV is a member of the BHV-4 group. In addition, the FeCAHV indirect FA test provides a sensitive and specific means of evaluating FECAHV antibody concentration in exposed cats.