Bronchoalveolar lavage was performed through an endotracheal tube in 34 specific-pathogen-free cats to determine expected values for bronchoalveolar lavage fluid cytologic analysis, using this method of collection. Saline solution for lavage was instilled in 3 separate aliquots at a volume of 5 ml/kg of body weight each. Analysis of sequential aliquots was performed to investigate the differences in cell counts among the 3 fractions. The effect of combining aliquots, including or omitting the first fraction, was evaluated to determine whether all aliquots could be combined for analysis without substantially affecting results. The total number of nucleated cells retrieved from each cat ranged from 0.9 to 31.1 X 10(6). Most of these cells were macrophages (78 +/- 15%, mean +/- SD) and eosinophils (16 +/- 14%). The first aliquot had the greatest number of epithelial cells, and the lowest total nucleated cell count and relative and absolute eosinophil counts. Differences among aliquots also were identified for relative and absolute macrophage counts, relative and absolute neutrophil counts, and absolute lymphocyte count. Statistically significant differences were found for many of the cell counts when values from the combination of the second and third aliquots were compared with values from the combination of all 3 aliquots. Magnitude of the differences was small, and these differences were not believed to be of practical consequence.

Effect of estrogen (E2) and progesterone (P4) on uterine antibacterial activity and immunoglobulin concentrations in mares was studied. In 2 in vitro experiments, 6 mixed-breed mares were ovariectomized, and uterine fluid and blood serum were analyzed. Antibacterial assay methods were used to determine inhibitory effects on Streptococcus zooepidemicus of uterine fluid samples collected on days 3, 5, and 8, and serum obtained on day 8 of treatment. Single radial immunodiffusion methods were used to quantify amounts of IgA and IgG in uterine fluid and serum on days 3, 5, 8, and 14 of treatment. Neither E2 nor P4 increased activity of serum and uterine fluid against S zooepidemicus. Numbers of colony-forming units per milliliter of bacteria were significantly (P < 0.01) lower in control Hanks' balanced salt solution with 1.0% gelatin (HBSSG) than in uterine fluids. Bacterial numbers were significantly (50%) greater in uterine fluids and serum than in HBSSG controls for both treatments. Both fluids, especially serum, supported significantly (P < 0.01) more growth of S zooepidemicus than did HBSSG when incubated for 0, 2, and 4 hours. These findings are in contrast to previous reports of antibacterial activity in the uterus of sexually intact mares undergoing an estrous cycle: great reduction of bacterial count in uterine fluid from mares in diestrus, and significant increases in bacterial numbers in uterine fluid or serum from mares in estrus. Treatment comparisons between serum and uterine fluid IgA and IgG concentrations were not significantly different, although overall IgA concentration in the uterus was higher than concentration in serum. The IgG concentration in uterine fluid was higher in P4- than E2-treated mares. However, IgG concentration was significantly (P < 0.01) higher in uterine fluid on day 8 in P4-treated mares than on day 3 or 5. Results of this study indicate that neither immunoglobulin concentration nor hormone treatment has a direct effect on streptocidal activity.

Cerebrospinal fluid and serum were obtained from 17 adult, healthy llamas (9 males, 1 castrated male, and 7 females). Osmolality; activities of lactate dehydrogenase and creatine kinase; and concentrations of glucose, sodium, chloride, potassium, total protein, and albumin were determined in serum and CSF. Total and differential cell counts were determined in CSF, and electrophoresis of CSF proteins was performed. Total nucleated cell count was low, 0 to 3/microliter, which is lower than that reported for other domestic species and is similar to values in healthy people. Differential leukocyte percentages were disparate depending on the degree of blood contamination. Blood contamination influenced the percentage of neutrophils and eosinophils in CSF. Samples with few erythrocytes had differential leukocyte distribution similar to that of other species: mostly lymphocytes, fewer monocytoid cells, and scant neutrophils. Older llamas had a few eosinophils in the CSF. Total protein, albumin, and gamma-globulin concentrations in llamas were similar to values in cattle and were higher than values in most domestic species. Glucose concentration in CSF was approximately 40% of the value in serum (nonruminant animals and people typically have CSF glucose concentration that is approximately 60 to 80% of the serum glucose concentration). Sodium and Cl concentrations in CSF were higher than those in serum, whereas K concentration was lower in CSF, compared with serum. Activities of creatine kinase and lactate dehydrogenase in CSF were markedly lower than those in serum, and the ranges of values in this group of healthy llamas were narrow.

Responses of the skin over the dorsum to solar UV irradiation (2 hours/d for 6 consecutive days) were investigated in hairless descendants of Mexican hairless dogs. Assessment of skin color changes, using a spectrophotometer, indicated that luminance values began to decrease from the third, day of UV irradiation, reached the minimal value at 3 weeks, and almost recovered 12 weeks after completion of UV irradiation. The number of the dihydroxyphenylalanine-positive melanocytes increased significantly (P < 0.01) from the third day of UV irradiation, reached its maximal value at 2 weeks, and recovered to normal at 12 weeks after completion of UV irradiation. On the second day of UV irradiation, the epidermis became focally thick, with disarrangement of component cells that had degenerative changes. In addition, a few so-called sunburn cells with pyknotic nuclei were seen in the epidermis. On the third day of UV irradiation, apparent suntan reaction developed, and a large number of epithelial cell in the epidermis were heavily pigmented with melanin granules. At 12 weeks after completion of UV irradiation, the epidermis appeared almost normal. On the other hand, significant changes were not detected in the dermis throughout the study.

Seven horses (4 anesthetized and 3 awake) and 2 ponies (anesthetized) were studied to evaluate the high sensitivity of the pulmonary circulation of the horse to various blood-borne particles, and to establish the presence of intravascular macrophages in the lung. Pulmonary and systemic pressures and cardiac output before and during particle injection were measured in some animals. An anesthetized foal had a large increase in pulmonary arterial pressure (32 and 34 mm of Hg) within 1 minute of iv administration of small test doses of radioactively labeled liposomes (2.5 micromoles/kg of body weight) or a 1% suspension of blue pigment (0.3 ml/kg), respectively. Quantitative real-time gamma camera imaging of the foal revealed high retention of the labeled liposomes during the first pass through the lungs; retention persisted throughout the experiment. Postmortem analysis revealed 55 and 47% lung retention of liposomes and blue pigment, respectively. The 2 anesthetized ponies had increased pulmonary artery pressure of 34 +/- 7 mm of Hg, decreased cardiac output, and 42% lung retention after administration of 1% blue pigment (0.2 ml/kg), whereas 3 awake horses had increased pressure of 28 +/- 9 mm of Hg after 1.8 X 10(8) (1.8-micromoles-diameter) latex microspheres/kg. None of the injected particles caused vascular obstruction, and they do not cause pulmonary vascular reactivity in species that lack pulmonary intravascular macrophages. Finally, 3 horses (1 anesthetized and 2 awake) were infused iv with small doses of the blue pigment, and their lungs were perfusion-fixed to identify specific labeling of the pulmonary intravascular macrophages. These cells were fully differentiated macrophages, contained blue pigment in phagocytes, and were tightly adherent to the pulmonary capillary endothelium. At this time, horses (order Perissodactyla) are the only species outside the mammalian order Artiodactyla (sheep, pig, cattle) documented to have reactive intravascular macrophages. Compared with other species, low doses of particles induced marked hemodynamic responses; horses appear to be more sensitive to IV administered particles than are other species studied.

Alterations in the size and functions of porcine alveolar macrophages exposed to sublytic amounts of heat-labile, hemolytic cytotoxin produced by Actinobacillus pleuropneumoniae (App) serotype 1, strain HS54 into the culture medium were studied in vitro. Alveolar macrophages were sensitive to the cytotoxin; treatment of the macrophages with low concentrations of cytotoxin (0.016 hemolytic unit) resulted in severe, irreversible cell swelling. However, high doses of cytotoxin (2.0 hemolytic units) were required to cause substantial cell death, as indicated by the influx of propidium iodide into and release of lactate dehydrogenase from cells. Macrophages exposed to low, sublytic doses of cytotoxin failed to migrate toward chemoattractant, were unable to attach to glass, and failed to phagocytize optimally opsonized erythrocytes. Macrophages already attached to glass surfaces detached when exposed to sublytic doses of cytotoxin. The swelling and impairment of functions of alveolar macrophages observed in this study could not be attributed to endotoxic effects, because heat treatment of the cytotoxin preparation for 60 minutes at 60 C resulted in complete loss of cytotoxicity. We conclude that sublytic doses of heat-labile, hemolytic cytotoxic substances produced by App depress alveolar macrophage function at concentrations likely to develop in association with acute pulmonary infection with App. The Apx (A pleuropneumoniae Rtx toxins) exotoxins secreted by the bacteria into culture medium were considered responsible for the toxic activity of the cytotoxin preparation. The Apx of the App field strain used in this study were likely to be similar to those of serotype-1 reference strain (S4707). Analysis by use of DNA-DNA hybridization indicated that genomic DNA of the field strain contained sequences similar to those encoding structural protein of ApxI (apxIA) and ApxII (apxIIA) of the serotype-1 reference strain. Therefore, Apx produced by the field strain of App used in this study are likely to be of similar pathogenic importance worldwide.

To determine the effects of age on each analyte, CSF variables were evaluated in healthy foals from birth through 42 days of age. Cerebrospinal fluid was collected from 14 clinically normal, naturally delivered cross-bred foals and was analyzed for glucose, sodium, potassium, magnesium, and total protein concentrations, total and differential WBC counts, RbC count, and lactate dehydrogenase, aspartate transaminase, and creatine kinase activities. Samples were collected in 3 foals < 48 hours old, and at 11 to 14 days of age in 4 foals, 21 to 22 days of age in 3 foals, and 31 to 42 days of age in 4 foals. Each foal was tested only once, to avoid any effects of CSF sample collection on subsequent analysis. Regression analysis confirmed age-related effects on CSF glucose, protein, and magnesium concentrations, but did not indicate an effect of age on CSF sodium and potassium concentrations or cell counts. Results indicate that CSF glucose concentration decreases with age; foals < 2 days old had the highest CSF glucose values, 98.8 +/- 12.0 mg/dl (mean +/- 1 SD). In foals 10 to 14 days old, CSF glucose concentration was 67.3 +/- 12.0 mg/ dl, was 65.3 +/- 4.5 mg/dl in foals 21 to 22 days old, was 70.0 +/- 5.4 mg/dl in foals 31 to 42 days old, and was 51.1 +/- 2.5 mg/dl in adults. Protein values in CSF also decreased with age: 109.0 +/- 9.7 mg/dl in foals < 2 days old, 81.0 +/- 22.8 mg/dl in foals 10 to 14 days old, 60.5 +/- 22.4 mg/dl in foals 21 to 22 days old, and 58.5 +/- 17.0 mg/di in foals 31 to 42 days old. The CSF protein concentration was 60.3 +/- 10.8 mg/dl in adult horses. Magnesium concentration in CSF increased slightly with age, then decreased after 22 days of life. In foals < 2 days old, the value was 2.43 +/- 0.16 mg/dl. Values in older foals and horses were: 2.51 +/- 0.08 mg/dl in foals 10 to 14 days old, 2.65 +/- 0.05 mg/dl in 21- to 22-day-old foals, 2.55 +/- 0.05 mg/dl in 31- to 42-day-old foals, and 2.35 +/- 0.09 mg/dl in adult horses. Mean CSF sodium and potassium concentrations were 151.7 +/- 3.7 mmol/L and 3.14 +/- 0.54 mmol/L, respectively, for all ages. There was no effect of age on these analytes. Values for CSF enzymes were considered invalid for the assay technique used and were not further analyzed.

The effect of right paralumbar fossa exploratory celiotomy and omentopexy on peritoneal fluid constituents was studied in 22 adult dairy cows. Six cows were eliminated on the basis of physical examination findings (n = 2), surgical findings (n = 2), or inability to obtain a sufficient volume of peritoneal fluid (n = 2). Sixteen cattle had normal results of Csc and serum biochemical analysis, and a minimum of 1 ml of peritoneal fluid was obtained by abdominocentesis. Abdominocentesis was repeated on days 1, 2, and 6 after surgery. Statistical analysis for repeated measures was performed, using a significance level of P < 0.05. Stage of gestation was evaluated for interaction with time. Mean total nucleated cell count was 3,200 cells/1 before surgery, was significantly increased 2 days after surgery (16,336 cells/microliter), and continued to increase through day 6 (20,542 cells/microliter). Mean polymorphonuclear cell count was 1,312 cells/microliter before surgery and was significantly higher at 2 (11,043 cells/microliter) and 6 (10,619 cells/microliter) days after surgery. Mean lymphocyte count was 254 cells/microliter before surgery and was significantly increased 2 days (1,911 cells/microliter) after surgery. By day 6, lymphocyte numbers were similar to preoperative values. Mean mononuclear cell count was 770 cells/microliter before surgery and was significantly increased on days 1 (3,084 cells/microliter), 2 (3,285 cells/microliter and 6 (2,349 cells/microliter) after surgery. Mean eosinophil numbers were 1,388 cells/microliter before surgery and were significantly increased on day 6 (6,347 cells/microliter) only. Interaction between time and stage of gestation was found only for specific gravity and total protein concentration. In general, specific gravity and total protein concentration increased after surgery (mean before surgery, 1.016 and 3.6 g/dl; mean after surgery, 1.021 and 5.6 g/dl). Left paralumbar fossa celiotomy performed 7 days after surgery did not reveal complications of repeated abdominocentesis, and pregnancy status was unchanged. Peritoneal fluid constituents are highly variable after exploratory celiotomy and omentopexy in cattle. However, results of this study may provide a reference for interpretation of postoperative peritoneal fluid sample findings in cattle.

The neutrophil plasma membrane has a major role in migration, phagocytosis, and destruction of microorganisms. Neutrophils isolated from blood and mammary secretions were homogenized, and the plasma membrane fraction was isolated on discontinuous sucrose gradient (20, 32, and 50%). Purity of plasma membrane preparation was determined by use of marker enzyme analysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the membrane proteins was performed under reducing conditions for polypeptide characterization. The membrane proteins were also labeled with 125I externally, using 1,3,4,6-tetrachloro-3 alpha-6 alpha-diphenylglycouril, and proteins were separated by SDS-PAGE and autoradiographed. Compared with whole cell homogenate, the plasma membrane fraction obtained at the 20/32% interface was enriched for the marker enzymes, 5'-nucleotidase (16-fold), alkaline phosphatase (5.5-fold), and total phosphatase (26-fold). The membrane fraction had minimal specific activity for beta-glucuronidase (0.4-fold), compared with whole cell homogenate. Plasma membrane protein yield was about 500 micrograms/ 10(9) bovine blood neutrophils. The SDS-PAGE of plasma membrane proteins revealed 25 protein bands, of which 13 were major bands. There were 3 distinct bands (18, 36, and 65 kd) in the plasma membrane-enriched fraction (20/32 interface) that were not seen in other fractions (30/50% and pellet). Further, 125I-labeling identified 5 distinct protein bands (205, 140, 65, 35, and 30 kd). Blood and mammary neutrophils had similar polypeptide patterns, except that 36- and 65-kd bands were more prominent for blood neutrophils than for mammary neutrophils.

Bronchoalveolar lavage (BAL) was performed on 16 horses to determine whether it caused local or diffuse inflammation in the lungs. In 7 horses, BAL was performed in both lungs twice, 48 hours apart. Although total cell counts of the BAL samples did not change significantly, there were increased numbers and percentage of neutrophils in the second lavage fluid samples. In 5 horses, BAL was performed in 1 lung and repeated 48 hours later in the same lung and in the corresponding airway in the contralateral lung. The absolute cell count and percentage of neutrophils were significantly (P = < 0.05) increased in the second sample from the lung that was lavaged twice. In 4 horses, BAL was performed in 1 lung and 48 hours later, repeated in an adjacent airway to the first BAL site, and in the corresponding airway in the contralateral lung. Significant differences were not detected in the total or differential cell counts of the BAL fluid recovered at any time, except for an increase in neutrophil percentage in the second sample in the contralateral lung. The increased neutrophil percentage values were within the range of normal for healthy horses. Results of the study suggested that, in horses, BAL induces a localized pulmonary neutrophil influx that persists at least 48 hours and is characterized by a relative and absolute increase in the number of neutrophils in the lavage fluids.