The comparative metabolism of diisopropyl methylphosphonate in mink and rats.

This study reports the metabolism of carbon-14labeled diisopropyl methylphosphonate (DIMP) in mink and rats, undertaken to better understand the dose-related mortality reported for mink in a previous study. In both male and female mink and rats, DIMP was rapidly absorbed after oral administration; it was metabolized by a saturable pathway to a single metabolite, isopropyl methylphosphonate (IMPA), which was rapidly excreted, primarily in the urine (90%). Fecal radioactivity, also identified as IMPA, was 1.7-3.1% of the administered dose. Female rats had a slower rate of conversion of DIMP to IMPA and less total excretion of IMPA than male rats. Metabolism of DIMP administered intravenously was not very different from that given orally in both species. These data indicate that mink absorb, metabolize, and excrete DIMP (as IMPA) in a manner very similar to mice, rats, and dogs.

Hematologic and serum chemistry reference values for adult brown mink.

Hematologic and serum chemistry reference values were determined for 160 12-month-old brown untamed captive mink (Mustela vision). Blood was obtained by jugular venipuncture after administration of ketamine and xylazine. There were no statistically significant differences between male and female mink. The packed cell volume, hemoglobin, and red blood cell count were 10 to 20% lower than previously reported for non-anesthesized mink. Serum glucose, alanine aminotransferase and aspartate aminotransferase values also were lower than previously reported values.

Hematologic response of adult brown mink to oxidative stress.

We evaluated the response of mink to oral administration of the oxidant compound propylene glycol (PG) to better understand the relative susceptibility of mink red blood cells (RBC) to oxidant injury. Feeding a diet containing 12% PG to 6 mink for 1 w resulted in a 17% decrease in hematocrit, a 21% decrease in RBC count and a 4.8-fold increase in reticulocyte count. A marked increase in Heinz body and eccentrocyte numbers was consistent with oxidative injury to RBC. Because of high food intake, mink ingested approximately twice the quantity of PG/kg body weight compared to domestic cats fed diets containing 12% PG. Therefore, the severity of the hematologic dyscrasia in mink may be the result of greater intake of PG rather than unique sensitivity of mink RBC of oxidative injury. However, the high food intake and the mink's position at the top of the food chain may increase its exposure to environmental contaminants.

Subchronic oral toxicity study of diisopropyl methylphosphonate in mink.

Diisopropyl methylphosphonate (DIMP), produced during manufacture of the chemical agent GB (Sarin), is a groundwater contaminant at Rocky Mountain Arsenal, Colorado. DIMP was fed for 90 days to dark brown "Ranch Wild" mink housed under controlled indoor conditions. One-year-old mink, 10 of each sex, were fed 0, 50, 450, 2700, 5400, or 8000 ppm in standard ranch diet. Actual DIMP consumption was 0, 8, 73, 400, 827, and 1136 mg/kg body wt/day, respectively. Two additional groups of 10 served as "pair-fed" controls. Body weight and food intake were recorded weekly. Complete blood count and 15 chemical analytes were measured at Weeks 0, 3, 7, and 13. Necropsy and microscopic examination were performed on all mink. No clinical morbidity or deaths occurred. Both sexes fed 8000 ppm ate approximately 20% less and weighed approximately 20% less than the controls; 5400 ppm females had a 10% weight decrement. Plasma cholinesterase (ChE) decreased in the top three dose groups starting at Week 3. At 13 weeks, decrements were approximately 50% but returned to normal after 1 week without DIMP. Erythrocyte ChE was not reduced. Heinz bodies occurred in 10-15% of RBCs in 50% of 8000 ppm mink at 13 weeks, and 0.1-2.0% of RBCs in 25% at 2700 ppm. There were mild decreases in RBC count, hematocrit, and hemoglobin, and increases in reticulocyte count, at the 5400 and 8000 ppm doses. All recovered within 3 weeks after DIMP was withdrawn. The 8000 ppm group had marginal splenic hematopoiesis, histologically. No other treatment-related changes were noted. The 450 ppm dose was a clear no-effect level (approximately 73 mg DIMP/kg body wt/day). Compared to reports of similar studies of DIMP in rats and dogs, these mink displayed no unique species susceptibility.

Assessment of the hematopoietic system in ruminants.

The hemic system is examined to evaluate clinical signs referable to the hemic system, establish a data base, or monitor the course of disease processes. Clinical signs of hemic disorders include pale mucous membranes, icterus, hemorrhage, hemoglobinuria, and tachycardia. Compared to other species, ruminants have small red blood cells, respond to anemia by releasing stippled red blood cells, commonly become neutropenic during acute bacterial sepsis, and fail to develop marked neutrophilia or left shift in response to inflammation.

Hematological alterations in kittens induced by 6 and 12% dietary propylene glycol.

Soft-moist cat foods containing 6-13% propylene glycol (PG) induce Heinz body formation and decreased red blood cell (RBC) lifespan in adult cats in a dose dependent manner. Since kittens eat relatively more food/kg of body weight and must expand their blood volume in addition to replacing senescent RBC, the hematologic dyscrasia associated with consumption of PG-containing diets may be exaggerated. To test this hypothesis 21 kittens were divided into 3 groups of 7 each and fed diets containing, 0,6 or 12% PG for 13 w. A dose-related increase in Heinz bodies occurred in the 6 and 12% PG groups within 2 w and persisted throughout the study. Although only slight changes occurred in hematocrit, hemoglobin and red blood cell (RBC) count, punctate reticulocytes increased significantly in the 6 and 12% PG groups indicating accelerated erythropoiesis. Mean RBC survival was decreased in the 6 and 12% groups by 44% and 63% respectively when compared to the control group. The increase in reticulocyte count and reduction in RBC lifespan was greater than observed in adult cats. The greater effect in kittens may be due to greater PG intake and not to an inate susceptibility of kitten RBC to PG.

Hematologic alterations in adult cats fed 6 or 12% propylene glycol.

Cat foods containing propylene glycol (PG) induce Heinz body formation in feline erythrocytes. To further study the hematologic importance of dietary PG, 21 adult cats were allotted to 3 groups of 7 each and fed diets containing 0, 6, or 12% PG on a dry-weight basis. Cats fed PG had a dose-related increase in Heinz bodies within 2 weeks, and the increase persisted throughout the study. Although only slight changes occurred in PCV, hemoglobin concentration, and RBC count, punctate reticulocytes were significantly increased in the group fed 12% PG. Mean RBC survival was decreased in the groups fed 6 or 12% PG by 30 and 55%, respectively, compared with the control group. These data indicate that PG-containing diets cause a dose-dependent erythrocyte destruction, even when fed at concentrations as low as 6%.

Effects of propylene glycol-containing diets on acetaminophen-induced methemoglobinemia in cats.

Soft-moist cat foods contain 7 to 13% propylene glycol (PG) on a dry-weight basis. These diets induce Heinz body formation in feline RBC. In this study, we evaluated cats on a control diet and on a commercial diet containing 8.3% PG. All cats on the PG diet developed an increase in the number of circulating Heinz bodies. We then administered acetaminophen to cats on each diet to determine whether RBC from cats on PG diets were more susceptible to oxidant stress. Methemoglobin concentrations were significantly greater in cats in PG diets after acetaminophen administration. These data indicate that RBC from cats fed PG diets are more susceptible to oxidative stress.

Surface-activated bovine platelets do not spread, they unfold.

The present study has examined the response of bovine platelets to surface activation and compared it to the reaction of human cells. Human platelets react to surfaces by losing their discoid shape, extending pseudopods, converting to dendritic forms, and finally, spreading into thin films resembling pancakes. Bovine platelets do not spread, they unfold. Surface activation causes them to transform from discs to irregular, flattened shapes resembling dendritic platelets, but they are unable to fill in spaces between pseudopods, a step required for spreading. Bovine platelets lack the surface-connected open canalicular system (OCS), which serves as a reservoir of membrane for human platelet spreading. Its absence may be the major factor in the failure of bovine platelet spreading, but there are other possible factors. Circumferential microtubules are more resistant to disassembly in surface-activated bovine than human cells, and their stability as rings or fractured bundles may limit spreading. Actin filament assembly is similar in human and bovine platelets, but the organization is different. Human platelets form a peripheral weave of actin that expands the membrane between pseudopods. A peripheral weave does not form in surface-activated bovine platelets. The absence of the OCS and differences in cytoskeletal organization in bovine platelets may also affect spreading of the surface membrane. Fibrinogen-gold (Fgn-Au) probes added to spread human platelet move from pseudopods and the cell margin toward the center and concentrate in the OCS. Fgn-Au particles bind to surface-activated bovine cells, but move very little, or not at all. All of these factors may contribute to the inability of bovine platelets to react to surfaces by spreading like human cells, but absence of the OCS appears to be the major cause.

Contribution of propylene glycol-induced Heinz body formation to anemia in cats.

Propylene glycol (PG) is a common preservative and source of synthetic carbohydrates in soft-moist pet foods. Propylene glycol was fed to cats for 5 weeks at concentrations found in commercial diets (1.6 g/kg of body weight; 12% of diet on a dry-weight basis) and for 3 weeks at concentrations exceeding usual intake (8 g/kg; 41% of diet). There was a dose-dependent increase in Heinz body percentage to 28% in cats fed the low dose of PG and to 92% in cats fed the high dose. Erythrocyte half-life, measured using [14C]-cyanate hemoglobin (Hb), decreased significantly (P less than 0.05) by 18.8% and 60% in cats fed the low and high PG doses, respectively. The PCV in cats fed the low dose was unaffected, whereas cats fed the high dose had a mean (+/- SEM) decrease in PCV from 33.5 +/- 1.05% to 26.3 +/- 1.45%, accompanied by punctate reticulocytosis and bone marrow erythroid hyperplasia. A dose-dependent increase in iron pigment was found in the liver and spleen of all cats. In cats fed the low dose of PG, erythrocyte reduced glutathione concentration actually increased from 7.02 +/- 0.56 to 9.74 +/- 0.69 mumol/g of Hb, but decreased to 2.96 +/- 0.27 mumol/g of Hb in cats fed the high dose. There was no significant increase in methemoglobin concentration. These results indicated that PG cannot be considered innocuous even at concentrations consumed by cats eating commercial diets. Heinz body-induced acceleration of RBC destruction develops in a dose-dependent manner, so that cats with greater food intake, ie, lactating queens and nursing kittens, are at greater risk for development of PG-induced Heinz body hemolytic anemia.

Reassessment of cytologic values in canine cerebrospinal fluid by use of cytocentrifugation.

Using cytocentrifugation, nearly one fourth of canine cerebrospinal fluid (CSF) samples with cell counts in the normal range had abnormalities in cell type or morphologic features. Cerebrospinal fluid samples from 145 dogs with neurologic disorders were evaluated by use of this method. These results indicate that low hemacytometer counts in canine CSF should not be interpreted as normal. By increasing the detection of abnormalities in CSF, cytocentrifugation might improve diagnosis and treatment of canine neurologic disease.

Canine prostatic disease--comparison of ultrasonographic appearance with morphologic and microbiologic findings: 30 cases (1981-1985).

A retrospective analysis was made of 30 cases of canine prostatic disease, with the objective of identifying (via a prepubic approach) the 2-dimensional, gray-scale ultrasonographic appearance most often associated with the various spontaneous prostatic diseases. Ultrasonography was of value in characterizing the parenchymal architecture as normal vs focally hyperechoic and diffusely hyperechoic (associated with chronic inflammation and neoplasia) or focally hypoechoic or anechoic (either accompanied by distant enhancement), which was associated with retention cyst or abscess. Further specificity based only on abnormal echotexture was not possible. Ultrasonography facilitated the differentiation of radiographically identifiable prostatomegaly attributable to abscess or neoplasia from apparent prostatomegaly attributable to paraprostatic cyst. An imaging protocol consisting of distention retrograde urethrocystography and prepubic ultrasonography was recommended, as a distended bladder aided ultrasonographic identification of the prostate gland. In addition, the combination of urethral morphologic features and urethroprostatic reflux appearance complemented the ultrasonographic appearance for differentiation of prostatic abscess from prostatic carcinoma. A classification scheme for spontaneous canine prostatic disease combining germane imaging morphologic features with microscopic and microbiologic findings was proposed.

Canine prostatic disease--comparison of radiographic appearance with morphologic and microbiologic findings: 30 cases (1981-1985).

A retrospective analysis was made of radiographs from 24 clinically normal young adult male Beagles used in previously reported base-line research on the distal male urinary tract. The ventrodorsal and craniocaudal prostate gland dimensions were measured, as visualized on the lateral radiographic view, and expressed as a ratio of the distance between the cranial aspect of the public bone and the sacral promontory. In 17 of these 24 dogs, there was sufficient image clarity to permit confident measurement of prostatic dimensions. From these data, the largest acceptable prostatic dimension, as visualized on the lateral radiographic view, was 70% of the public-promontory distance. A retrospective analysis was made of 30 cases of canine prostatic disease, with the objective of identifying the radiographic (survey and distention retrograde urethrocystogram) appearance most often associated with the various spontaneous prostatic diseases. Prostatomegaly was not specific for the cause of prostatic disease, except if the apparent parenchymal dimensions were greater than or equal to 90% of the public-promontory distance wherein neoplasia, abscess, and paraprostatic cysts (readily differentiated from diffuse intraparenchymal disease by ultrasonography) were the principle considerations. Multifocal, irregularly shaped, parenchymal mineral densities were observed only in dogs with prostatic carcinoma (4/7 cases) or prostatitis (1 case of chronic prostatitis). Narrowing of the prostatic portion of the urethra was observed only in association with abscess, neoplasm, or noninflammatory nonneoplastic disease (possibly benign hypertrophy/hyperplasia), whereas widening of this part of the urethra was not observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytochemical reactions in cells from leukemic dogs.

Leukemic cells from 17 dogs with spontaneous leukemia were stained with leukocyte alkaline phosphatase, alpha naphthyl acetate esterase with and without fluoride, peroxidase, and periodic acid-Schiff. Cytochemistry was necessary for identification or confirmation of leukemic cell type in most dogs and resulted in changing the light microscopic morphologic diagnosis in eight of 17 dogs. Leukemic cell types diagnosed were myelomonocytic leukemia in seven dogs, monocytic leukemia in five dogs, lymphocytic leukemia in four dogs, and myelocytic leukemia in one dog.

Chemical reactions in cells from leukemic cats.

Cytochemical staining for leukocyte alkaline phosphatase(LAP), nonspecific esterase (NSE), nonspecific esterase with fluoride inhibition (NSE-F), periodic acid Schiff (PAS) reactivity, and peroxidase (PO) was valuable in identification of the neoplastic cell type in 10 leukemic cats. Staining both blood and bone marrow smears was often necessary for making the correct diagnosis. Cytochemical staining resulted in changing the morphologic diagnosis of leukemia in two of the 10 cats. Also, increased LAP activity, probably a marker for myelocytic leukemia in the cat, was observed in bone marrow cells from three nonleukemic, FeLV-positive cats.

Urate nephrolithiasis in ranch mink.

Urate nephrolithiasis with epistaxis and bleeding around foot pads was diagnosed in ranch mink. Investigation of affected, unaffected-related, and unaffected-unrelated mink did not disclose the cause of these problems but did eliminate inherited bleeding disorders as a cause of the bleeding. All affected mink were males, had been sired by related sires, and were affected in the 1st year of life. Elimination of affected mink and their parents from the breeding stock eliminated the condition from the herd. The findings were consistent with an inherited defect in uric acid metabolism or excretion.

A comparison of polychromasia and reticulocyte counts in assessing erythrocytic regenerative response in the cat.

Three staining techniques were evaluated for use in assessing the erythrocytic regenerative responses in the cat. Using new methylene blue as a vital stain, the aggregate reticulocyte count closely corresponded to the reticulocyte count on air-dried smears stained with new methylene blue and to the polychromatophilic erythrocyte count on Wright's stained smears.

Carboxyhemoglobin concentrations in normal cattle.

Percentage carboxyhemoglobin (HbCO) values were determined in 29 cows and 24 bulls in Minnesota during the summer of 1973. Results indicated that, in normal cattle, the HbCO value was 0.3%. Individual and daily variations in these values were recorded in 2 calves.