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Development of methods for analyzing plasma lipoprotein concentrations and associated enzyme activities and their use to measure the effects of pregnancy and lactation in cats
1995
Methods available for measurement of plasma lipoprotein-cholesterol concentrations and activities of lipoprotein lipase, hepatic lipase, lecithin:cholesterol acyl transferase (LCAT), and cholesteryl ester transfer protein were adapted for use in cats. A combined ultracentrifugation/precipitation procedure was used to isolate very low-density lipoproteins (VLDL), then to separate low-density lipoproteins (LDL) from high-density lipoproteins (HDL). The reagent used, 92 mM heparin-manganese chloride, provided complete precipitation of LDL with only trace and insignificant contamination by HDL. Efforts to selectively measure lipoprotein lipase activity in plasma, collected after IV injection of heparin, by inhibiting hepatic lipase with sodium dodecyl sulfate were unsuccessful, and the activity of this enzyme was calculated as the difference between total and hepatic lipase activities. The latter was measured in the presence of high salt concentration to inhibit lipoprotein lipase. Cholesterol esterifying activity was identified in feline plasma and was typical of LCAT, in that it was dependent on apolipoprotein A-I as a cofactor. The intra-assay and interassay coefficients of variation for measurement of lipoprotein lipase, hepatic lipase, and LCAT activities were 18.4, 4.6, and 7.2%, and 20.4, 10.7, and 5.3%, respectively. Appreciable cholesteryl ester transfer protein activity was not detected in either undiluted or diluted plasma. These methods were subsequently used to investigate the effects of pregnancy and lactation on lipoprotein metabolism in a group of 10 queens. Plasma concentrations of cholesterol and triglycerides were unaltered during pregnancy, but the concentrations of VLDL-cholesterol increased and those of HDL-cholesterol decreased. During lactation, the concentrations of cholesterol and triglycerides decreased owing to reductions in VLDL-cholesterol and LDL-cholesterol concentrations and continued suppression of HDL-cholesterol. These changes were associated with alterations in the activities of lipoprotein lipase, which increased after parturition, and hepatic lipase, which increased during pregnancy and lactation, that may help explain their metabolic origins. The activity of LCAT remained unchanged.
Afficher plus [+] Moins [-]Efficacy of domperidone and sulpiride as treatments for fescue toxicosis in horses
1994
Redmond, L.M. | Cross, D.L. | Strickland, J.R. | Kennedy, S.W.
We evaluated the effectiveness of 2 dopamine antagonists as treatments for fescue toxicosis in horses. Sixteen gravid mares were assigned by breed and expected foaling date to 1 of 3 treatment groups: endophyte-infested control 1.1 mg of domperidone/kg of body weight/d; and 3.3 mg of sulpiride/kg/d. Mares were pastured on endophyte-infected fescue and received 0.454 kg of a corn and dried molasses carrier containing the drug treatment. Treatment started 30 days prior to expected foaling date and continued until parturition. Blood samples were collected, and mammary gland scores were recorded every 5 days. Body weight and body condition scores were obtained every 28 days. Serum was analyzed for prolactin, progesterone, and estradiol-17beta concentrations. Domperidone-treated mares had shorter (P = 0.09) gestation duration and foaled closer (P = 0.07) to their expected parturition date than did control mares. Mammary gland scores were higher (P < 0.05) for domperidone-treated mares than for control mares. By 4 and 9 days after the start of treatment, serum prolactin concentration was higher P < 0.05) in domperidone-treated mares and sulpiride-treated mares, respectively, than in control mares. Domperidone- and and sulpiride-treated mares had higher (P < 0.05) serum progesterone and lower (P < 0.01) estradiol-17beta concentrations than did control mares. These results indicate that domperidone may offer considerable potential as a treatment for fescue toxicosis in horses.
Afficher plus [+] Moins [-]Effect of early lactation milk yield on reproductive disorders in dairy cows
1994
Grohn, Y.T. | Hertl, J.A. | Harman, J.L.
Association between individual cumulative milk yield and various reproductive disorders in 56,772 Finnish Ayrshire cows belonging to 5,912 herds in 80 communities was studied. All cows delivered calves between September 1985 and September 1986. Five logistic regression models were fitted, 1 for each outcome disorder of interest: early metritis, late metritis, silent heat, ovarian cyst, and other infertility. Cumulative individual 37-day milk yield was used in the early metritis model, and cumulative individual 60-day milk yield was used in the other models, on the basis of median days in milk when these disorders developed. Cumulative 305-day herd milk yield, parity, calving season, presence or absence of other disorders, and community were also included in the models. Point estimates from the models represented odds ratios for the likelihood of having the outcome disorder.Lactational incidence risks for the 5 reproductive disorders studied were: early metritis (2.4%), late metritis (1.1%), silent heat (5.4%), ovarian cyst (6.6%), and other infertility (2.1%). The risk of early metritis decreased with increasing 37-day milk yield. The risk of silent heat, ovarian cyst, and other infertility increased with increasing 60-day milk yield; 60-day milk yield had no effect on late metritis. The 305-day herd milk yield increased the risk of early metritis, ovarian cyst, and other infertility; it had no effect on late metritis or silent heat. Parity had an effect on all disorders, except late metritis. Cows that delivered calves during the colder, darker seasons of the year had a higher risk of reproductive disorders than did those that delivered calves at other times of the year. A number of other disorders, reproductive and otherwise, were significant predictors of development of the outcome disorders.
Afficher plus [+] Moins [-]Cytokine production during endotoxin-induced mastitis in lactating dairy cows
1993
Shuster, D.E. | Kehrli, M.E. Jr | Stevens, M.G.
The role of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor a during endotoxin-induced mastitis in cows was characterized. Six cows had 10 microgram of Escherichia coli lipopolysaccharide infused into 1 mammary gland. Three other cows served as nontreated controls. Within 1.5 to 2.5 hours after infusion, endotoxin caused obvious edema of the mammary gland and increased serum albumin concentration in milk of infused glands 6 times. Milk somatic cell count began to increase 3 to 5 hours after infusion in all treated glands. At 7 hours after infusion, somatic cell counts were increased > 10 times, compared with counts in milk from control cows. Pyrexia of > 1 C developed in only 1 cow, but all treated cows had serum cortisol concentrations > 50 ng/ml in response to endotoxin treatment. High concentrations of IL-1 (10 to 600 U/ml) and IL-6 (2 to 22 U/ml) were detected in milk of infused glands beginning 2.5 to 4 hours after infusion. Endotoxin did not induce detectable amounts of tumor necrosis factor activity in milk or serum. Swelling and mammary gland permeability changes preceded any detectable increase in IL-1 and IL-6 activity, indicating that these clinical signs of inflammation were not mediated by these cytokines. Systemic responses and the leukocytic influx into endotoxin-infused glands developed after or concurrently with initial increases in IL-1 and IL-6 activities in milk. These results suggested that IL-1 and IL-6 may have a role in mammary gland defenses and in the pathophysiologic changes during endotoxin-induced mastitis.
Afficher plus [+] Moins [-]Relation among body condition score, serum urea nitrogen and cholesterol concentrations, and reproductive performance in high-producing Holstein dairy cows in early lactation
1992
Ruegg, P.L. | Goodger, W.J. | Holmberg, C.A. | Weaver, L.D. | Huffman, E.M.
Body condition scoring (using a 5-point with quarter-point divisions) was performed on 66 Holstein dairy cows that began their second or later lactation in August, September, or October 1988. Cows' body condition was scored beginning on postpartum day 4 (+/- 1) and subsequently at postpartum days (+/- 1) 18, 32, 46, 60, 73 and 87. Blood samples were obtained on the same dates. Reproductive health examinations were conducted by 1 of 2 veterinarians beginning at postpartum day 21. Reproductive performance was evaluated in relation to body condition score and serum urea nitrogen and cholesterol concentrations. Number of days to first recorded signs of estrus and first breeding were not related to body condition score at calving, amount of condition loss, cumulative 80-day milk yield, or 305-day fat corrected milk yield. Cows that calved with body condition score greater than or equal to 3.50 required more days to conceive. Cows losing > 0.75 points of condition had longer days of conception. Body condition score at calving and amount of condition lost were not related to services per conception or diagnosis of follicular cyst. Cumulative 80-day milk yield was not related to days to conception or services per conception. Cows that produced greater than or equal to the mean 305-day milk yield required more services and had longer days to conception than cows that produced < the mean 305-day milk yield. Cows with diagnosis of ovarian follicular cysts had greater cumulative 80- and 305-day milk yields than did cows that were not diagnosed with follicular cysts. Cows conceiving with less than or equal to 2 services did not differ in average daily milk production, body condition score, or serum urea nitrogen concentration from cows conceiving with > 2 services, but cows that conceived with less than or equal to 2 services had higher serum cholesterol values than did cows requiring more services.
Afficher plus [+] Moins [-]Comparison of methods to determine glucose kinetics and measurement of recycling and lactose formation in lactating cows
1991
Biedebach, M.C. | Anand, R.S. | Black, Al | Jevning, R.
Several investigators have suggested that the conventional multicompartmental exponential analysis of in vivo glucose metabolism is arbitrary and possibly not the most accurate description of glucose kinetics, especially in the large animal. In support of that hypothesis, we found that in a systematic comparison of 3 methods, blood-specific radioactivity data in single-injection studies of glucose metabolism in lactating cows was better described graphically, or by a hybrid polynomial-biexponential curve fit, than by an exclusively exponential curve fit. We hypothesized that this finding was attributable to partial failure of linearity and steady-state assumptions that underlie the exponential model. Second, using both an irreversible tracer (3H-labeled glucose) and reversible tracer (14C-labeled glucose), we found that glucose carbon recycling had no effect during the first 2 hours, but became significant in lactating cows 7 hours after injection. Finally, we determined that approximately 52 to 55% of the glucose replacement rate was being used to generate lactose.
Afficher plus [+] Moins [-]Effects on aflatoxin M1 residues in milk by addition of hydrated sodium calcium aluminosilicate to aflatoxin-contaminated diets of dairy cows
1991
Harvey, R.B. | Phillips, T.D. | Ellis, J.A. | Kubena, L.F. | Huff, W.E. | Petersen, H.D.
Hydrated sodium calcium aluminosilicate (HSCAS), an anticaking agent for agricultural feeds, was added to aflatoxin (AF)-contaminated diets of 3 lactating dairy cows and evaluated for its potential to reduce aflatoxin M1 (AFM1) residues in milk. During phase I, cows were fed alternating diets that consisted of 200 microgram of AF/kg of feed for 7 days, 0.5% HSCAS plus 200 microgram of AF/kg of feed for 7 days, and feed with the HSCAS removed for a final 7 days. The AFM1 milk concentrations from the intervals with HSCAS added to diets were compared with those times when HSCAS was absent. The presence of 0.5% HSCAS in feed containing 200 microgram of AF/kg reduced AFM1 secretion into the milk by an average of 0.44 microgram/L (from pretreatment of 1.85 microgram/L to 1.41 microgram/L with HSCAS, a 24% reduction). Following a 10-day period of noncontaminated feed consumption and no AFM1 residues in the milk, phase II of the study was begun. The same experimental design as phase I was used, but the dosages of HSCAS and AF were changed to 1.0% and 100 microgram/kg of feed, respectively. The addition of 1.0% HSCAS in feed containing 100 microgram of AF/kg decreased AFM1 content in the milk by an average of 0.40 microgram/L (from a pretreatment of 0.91 microgram/L to 0.51 microgram/L when HSCAS was present, a 44% reduction). These findings suggest that HSCAS, a high-affinity sorbent compound for AF in vitro, is capable of reducing the secretion of AFM1 into milk.
Afficher plus [+] Moins [-]Effect of milk stasis on Brucella abortus infection of the mammary gland in goats
1991
Meador, V.P. | Deyoe, B.L.
To compare the effects of milk stasis and milk flow on Brucella abortus infection of the mammary gland under the same systemic conditions, primiparous goats (n = 5) were inoculated IV with B abortus on the day of parturition, and suckling by their neonates was restricted to one mammary gland. Goats were euthanatized and necropsied at 3 weeks after inoculation, and milk, mammary glands, and supramammary lymph nodes were evaluated by bacteriologic, histologic, and immunoenzymatic staining techniques. Nonnursed mammary glands had high titers of brucellae in milk, moderate interstitial mastitis, and brucellar antigen in macrophages located primarily in alveolar and ductal lumina. Brucellae often filled the macrophage cytoplasm. In contrast, nursed mammary glands had fewer brucellae in milk, minimal inflammatory changes, and no detectable brucellar antigen in histologic sections. Hyperplastic changes were only seen in supramammary lymph nodes draining nonnursed mammary glands; these contained more brucellae than lymph nodes draining nursed mammary glands. These studies show that milk stasis may be the sole cause of increased susceptibility of nonnursed mammary glands to B abortus infection.
Afficher plus [+] Moins [-]Pharmacokinetics of flunixin meglumine in lactating cattle after single and multiple intramuscular and intravenous administrations
1990
The pharmacokinetics of flunixin were studied in 6 adult lactating cattle after administration of single IV and IM doses at 1.1 mg/kg of body weight. A crossover design was used, with route of first administration in each cow determined randomly. Plasma and milk concentrations of total flunixin were determined by use of high-pressure liquid chromatography, using an assay with a lower limit of detection of 50 ng of flunixin/ml. The pharmacokinetics of flunixin were best described by a 2-compartment, open model. After IV administration, mean plasma flunixin concentrations rapidly decreased from initial concentrations of > 10 micrograms/ml to nondetectable concentrations at 12 hours after administration. The distribution phase was short (t1/2 alpha, harmonic mean = 0.16 hours) and the elimination phase was more prolonged (t1/2 beta, harmonic mean = 3.14 hours). Mean +/- SD clearance after IV administration was 2.51 +/- 0.96 ml/kg/min. After IM administration, the harmonic mean for the elimination phase (t1/2 beta) was prolonged at 5.20 hours. Bioavailability after IM dosing gave a mean +/- SD (n = 5) of 76.0 +/- 28.0%. Adult lactating cows (n = 6) were challenge inoculated with endotoxin as a model of acute coliform mastitis. After multiple administration (total of 7 doses; first IV, remainder IM) of 1.1 mg/kg doses of flunixin at 8-hour intervals, plasma flunixin concentrations were approximately 1 microgram/ml at 2 hours after each dosing and 0.5 microgram/ml just prior to each dosing. Flunixin was not detected in milk at any sampling during the study. Flunixin concentrations necessary to induce therapeutic effects in cattle are unknown. Results of our study indicate that administration of 1.1 mg/kg doses of flunixin meglumine at 8-hour intervals would produce plasma concentrations similar to those demonstrated to be effective clinically in treatment of equine musculoskeletal disorders and colic.
Afficher plus [+] Moins [-]Evaluation of Haemonchus contortus infections in sexually intact and ovariectomized ewes
1988
Fleming, M.W. | Rhodes, R.C. III. | Gamble, H.R.
Three experiments were conducted to investigate experimentally the occurrence of periparturient nematode egg rise in ewes and the hormonal modulation of Haemonchus contortus infections. In the first experiment, fall-bred and winter-bred pregnant (n = 4 and 14, respectively) and nonpregnant (n = 5 and 29, respectively) ewes were treated with anthelmintic and were pastured together on fields that were contaminated with H contortus. Three weeks before lambing, all ewes were placed in concrete pens; fecal egg counts for the winter-bred group were obtained on alternate days. Pregnant and lactating ewes had significantly larger numbers (P < 0.01) of H contortus eggs than did the nonpregnant controls 1 week before and after lambing. Lactating, fall-bred ewes had significantly (P < 0.01) more adult worms in their abomasum through natural acquisition than the nonpregnant controls. In the second experiment, fall-bred and winter-bred helminth-free, pregnant (n = 4 and 8, respectively) and nonpregnant (n = 3 and 15, respectively) ewes were inoculated on 5 alternate days, beginning 70 days after breeding with 20,000 infective H contortus larvae. The ewes were maintained on concrete pens throughout pregnancy. Fecal egg counts were significantly (P < 0.05) greater in pregnant ewes, beginning 1 week before lambing until 1 week after lambing. Abomasums of lactating ewes from both lambing seasons yielded significantly (P < 0.01) more adult worms at necropsy than nonpregnant ewes. In the third experiment, ewes were ovariectomized (n = 15) or sham-operated (n = 9); half of the control ewes were bred. Beginning on day 70 of pregnancy, all ewes were inoculated orally with 20,000 infective H contortus larvae on 5 alternative days. Abomasums were removed from all ewes after lambing, and adult worms were recovered. Pregnant ewes and half of the ovariectomized ewes had significantly (P < 0.05) more worms than did the sham-operated ewes.
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