Over a 54-hour period, blood was removed from 8 adult sheep (body weight, 38.1 +/- 0.5 kg mean +/- SEM) in 9 episodes, 5 on day 1, 3 on day 2, and 1 on day 3. Cumulative blood loss was 1,630 +/- 63, 2,380 +/- 71, and 2,693 +/- 69 ml on days 1, 2, and 3, respectively. Blood samples (20 ml) were collected from 5 control ewes (33.8 +/- 2.8 kg) at equivalent times. Over the first day, mean arterial blood pressure decreased in the hemorrhaged sheep from 101 +/- 2 mm of Hg to 76 +/- 5 mm of Hg, but returned to control values by the beginning of the second day and, thereafter, was not different from control values. Heart rate was increased after the first hemorrhage episode and remained high throughout the entire protocol. Over the entire period, there were statistically significant decreases in hematocrit, plasma osmolality, sodium, total calcium (P < 0.001), potassium, and chloride values (P < 0.05). There was no change in plasma phosphate, bicarbonate, creatinine, or magnesium concentrations and an increase in plasma urea nitrogen (P < 0.001) concentrations. Plasma arginine vasopressin concentration was increased significantly (P < 0.001) over the entire period. Plasma ACTH concentration was significantly (P < 0.05) increased over time, but only some values on day 1 were significantly outside the normal range of the control group data. Because of wide variation between sheep, the group data for aldosterone were not significantly different from control values. Blood volume was restored on day 1 with fluid of osmolality, Na, and Cl composition equivalent to that of plasma. The effects of arginine vasopressin were apparent by day 2, when the major decrease in osmolality and Na and Cl concentrations were observed The sheep has good capacity to withstand severe, prolonged hemorrhage, most likely because of a large reserve of RBC in the spleen; hematocrit remained at 31% of control values when an estimated 100% of initial circulating blood volume had been removed.

Polyclonal rabbit antiserum to human T-cell CD3 was used to study its reactivity in lymphoid tissues (lymph nodes, spleen, aggregated lymphoid follicles [Peyer's patches], thymus) of several animal species (cattle, sheep, goats, rats, and mice). Using a peroxidase-antiperoxidase technique on formalin-fixed and paraffin-embedded tissues, immunoreactive cells were detected in T cell-dependent areas of the lymphoid tissues. Reactivity was high in all species tested, but mouse tissues had reduced reactivity, compared with the other species. To obtain a reaction, it was necessary to digest tissues with pronase before application of the immunocytochemical technique. Our results indicate that CD3 antiserum may specifically recognize T-lymphoid cells as it does in human lymphoid tissues and can be used as a marker to study physiologic and pathologic conditions of the lymphoid system of these species.

An agar gel immunodiffusion (AGID) test was used over a 3-year period to examine 1,871 serum samples from sheep representing 5 Mycobacterium paratuberculosis infected flocks and 4 flocks presumed to be uninfected. Of 1,032 sheep, 31 had positive AGID test results (scoring 1 to 5), and 23 of these 31 were ecropsied. Infection with M paratuberculosis was confirmed by 1 or more of the following findings: observation of typical lesions on histologic examination of sections of ileum or ileocecal lymph nodes, observation of clumps of acid-fast bacteria in mucosal smears of ileum, and isolation of the organism from feces or tissue. False-positive results on AGID testing were not found in sheep from flocks known to have exposure to Cotynebacterium pseudotuberculosis. Diarrhea in infected sheep was observed infrequency; chronic, severe weight loss was the most common sign observed. On histologic examination of tissues from 20 infected sheep, 16 (80%) had diffuse lesions of the ileum and 13 (65%) had acid-fast bacteria in areas of ileal inflammation; 4 had discrete granulomas and peripheral lymphocytic infiltrates in the ileum. Sheep with diffuse lesions tended to have higher mean scores on AGID testing and examination for acid-fast bacteria, compared with those from sheep with more discrete lesions. Bacteriologic culture yielded M paratuberculosis from only 3 sheep with paratuberculosis. On the basis of results of this study, we suggest that the nature of the response to infection with M paratuberculosis may influence the results of diagnostic tests for paratuberculosis, and that AGID testing may be useful to identify M paratuberculosis infection in sheep with chronic weight loss and in flock-screening programs.

Corynebacterium pseudotuberculosis phospholipase D (PLD) significantly affected viability of ovine neutrophils after 24 hours' exposure, This effect was more marked in cells that ingested PLD emulsified in oil. Treatment of neutrophils with PLD significantly (P < 0.05) reduced the ability of these cells to migrate toward activated sheep serum. The PLD was not chemotactic, but it activated normal sheep serum, producing factors that were chemotactic for neutrophils.

Previous studies of the amino acid analogue, alpha-ketoisocaproate (KIC), indicate that it can stimulate lymphocyte blastogenesis and antibody responses of sheep. To determine whether KIC could overcome the effects of adreno-corticotropic hormone (ACTH)-induced lymphocyte suppression, 24 lambs were fed a control diet, a diet supplemented with 0.05% KIC, or a diet supplemented with 0.05% of the parent amino acid leucine. Immune status was monitored by determining lymphocyte blastogenic responsiveness to phytohemagglutinin-P (PHA), concanavalin A (conA), and pokeweed mitogen (PWM) and percentages of T-cell subsets in the blood, using monoclonal antibodies and a flow cytometer. Serum cortisol, insulin, and glucagon concentrations also were determined. After 60 days of consuming the respective diet, lambs were administered either saline solution or ACTH (100 IU) twice daily for 3 consecutive days. Administration of ACTH increased serum cortisol and insulin concentrations; however, no effects were seen for serum glucagon concentration. Compared with saline administration, ACTH administration significantly (P < 0.05) suppressed mitogen-stimulated lymphocyte blastogenesis by approximately 50%, regardless of the mitogen used, and significantly (P < 0.01) decreased the percentage of circulating T lymphocytes and decreased (P < 0.01) the ratio of T4 to T8 cells. Lambs fed KIC had greater PHA- and conA-stimulated blastogenic responses and significantly (P < 0.05) increased ratio of T4 to T8 cells in the blood, compared with lambs fed the leucine-supplemented diet or the control diet and given corresponding injections. These data indicate that ACTH decreased in vitro lymphocyte blastogenesis and altered the subset ratios of blood lymphocytes in sheep. These changes were partially prevented by feeding KIC.

In a survey of 666 sheep at a slaughterhouse, gallstones (concretions with a diameter greater than or equal to 1 mm) were found in the gallbladder of 50 sheep (7.5%), sludge (concretions with a diameter < 1 mm) was found in 9 sheep (1.4%), and sludge plus gallstones were found in 7 sheep (1.1%). Gallstones and sludge were associated, and were more frequent in lambs and females, compared with adults and males. Qualitative analysis of the stones revealed all to be pigment (bilirubin) stones. There was a statistically significant increase of biliary bilirubin (total and indirect quota) only in sheep with gallstones plus sludge, compared with control sheep without sludge or gallstones. Concentrations of bilirubin, cholesterol, phospholipids, total and single bile aids, and total and ionized calcium were similar in the bile of sheep with gallstones, sludge, or both and control sheep. Bacteriologic analysis of the bile in 10 sheep with gallstones and 10 controls revealed bacteria in 50% of the first group and in 75% of the second group (Escherichia coli in all sheep and Salmonella spp also in 1 sheep with gallstones). These findings confirm our earlier findings of a high prevalence of black pigment gallstones in sheep. On that basis, we suggest that gallstones are associated with high total bilirubin concentration in the bile, and deconjugating bacteria are common in the biliary tract of these animals.

Efficacy of oral administration of 20 mg of triclaben-dazole/kg of body weight was evaluated against 12-week Fascioloides magna infections in 12 sheep, each inoculated orally with 250 viable metacercariae. From 6 sheep treated with triclabendazole, 1 immature F magna was recovered, whereas 116 F magna with a mean length of 19 +/- 6.5 mm were recovered from 6 untreated control sheep. Efficacy of triclabendazole was 99.14%. Signs of toxicosis or illness were not observed in the sheep.