Changes in fetal mannose and other carbohydrates induced by a maternal insulin infusion in pregnant sheep

Background

The importance of non-glucose carbohydrates, especially mannose and inositol, for normal development is increasingly recognized. Whether pregnancies complicated by abnormal glucose transfer to the fetus also affect the regulation of non-glucose carbohydrates is unknown. In pregnant sheep, maternal insulin infusions were used to reduce glucose supply to the fetus for both short (2-wk) and long (8-wk) durations to test the hypothesis that a maternal insulin infusion would suppress fetal mannose and inositol concentrations. We also used direct fetal insulin infusions (1-wk hyperinsulinemic-isoglycemic clamp) to determine the relative importance of fetal glucose and insulin for regulating non-glucose carbohydrates.

Results

A maternal insulin infusion resulted in lower maternal (50%, P < 0.01) and fetal (35-45%, P < 0.01) mannose concentrations, which were highly correlated (r² = 0.69, P < 0.01). A fetal insulin infusion resulted in a 50% reduction of fetal mannose (P < 0.05). Neither maternal nor fetal plasma inositol changed with exogenous insulin infusions. Additionally, maternal insulin infusion resulted in lower fetal sorbitol and fructose (P < 0.01).

Conclusions

Chronically decreased glucose supply to the fetus as well as fetal hyperinsulinemia both reduce fetal non-glucose carbohydrates. Given the role of these carbohydrates in protein glycosylation and lipid production, more research on their metabolism in pregnancies complicated by abnormal glucose metabolism is clearly warranted.     

Pharmacokinetics of tulathromycin in fetal sheep and pregnant ewes

Macrolides are important antimicrobials frequently used in human and veterinary medicine in the treatment of pregnant women and pregnant livestock. They may be useful for the control of infectious ovine abortion, which has economic, animal health, and human health impacts. In this study, catheters were surgically placed in the fetal vasculature and amnion of pregnant ewes at 115 (±2) days of gestation. Ewes were given a single dose of 2.5 mg/kg tulathromycin subcutaneously, and drug concentrations were determined in fetal plasma, maternal plasma, and amniotic fluid at 4, 8, 12, 24, 36, 48, 72, 144, and 288 hr after drug administration. Pharmacokinetic parameters in maternal plasma were estimated using noncompartmental analysis and were similar to those previously reported in nonpregnant ewes. Tulathromycin was present in fetal plasma and amniotic fluid, indicating therapeutic potential for use against organisms in these compartments, though concentrations were lower than those in maternal plasma. Time‐course of drug concentrations in the fetus was quite different than that in the ewe, with plasma concentrations reaching a plateau at 4 hr and remaining at this concentration for the remainder of the sampling period (288 hr), raising questions about how tulathromycin may be transported into or metabolized and eliminated by the fetus.     

Persistent hypoglycemia induced by continuous insulin infusion in broiler chickens

1. Persistent hypoglycaemia was experimentally induced by insulin infusion to improve understanding of the regulatory mechanisms of blood glucose concentrations specific to chickens. 2. An osmotic minipump containing bovine insulin was implanted to deliver insulin in vivo at a constant rate (11.25 to 45 U/kg BW/d) for 5 d in 4-week-old broiler chickens force-fed a maintenance diet once a d. Birds infused with the highest dose of insulin died within 3 to 4 d. 3. In chickens continuously infused with insulin at 22.5 U/kg BW/d, fasting glucose concentrations in plasma determined every 3 h during the 3rd day of infusion were consistently and significantly lower than in controls. 4. Continuous infusion of insulin at 22.5 U/kg BW/d induced persistent hypoglycaemia (almost one-half the normal blood glucose concentration) lasting for at least 4 d in broiler chickens. 5. Insulin infusion did not significantly change plasma NEFA or protein concentrations and increased plasma GOT activity only at 1 of the daily experimental sampling points.     

Utilization of magnesium and other macrominerals in sheep supplemented with different readily-fermentable carbohydrates

Sheep were used to determine the effects of dietary supplementation with readily-fermentable carbohydrates on Mg, Ca, K and P utilization. In each of two metabolism trials, 15 mature, crossbred wethers (average weight, 49.2 kg) were allotted to five dietary treatments consisting of 800 g/d of orchardgrass (Dactylis glomerata, L.) hay alone, or supplemented with 450 g/d of either glucose, sucrose, lactose or starch. Each trial consisted of a 5-d adjustment period, a 10-d preliminary period and a 10-d collection period. Compared with wethers fed hay alone, supplementation with each kind of carbohydrate to the diet decreased (P less than .05) fecal Mg excretion and increased (P less than .05) apparent absorption and retention of Mg. Apparent absorption of the Ca was lower (P less than .05) in wethers fed lactose and tended to be decreased by glucose, sucrose and starch supplementation. Calcium retention was lower (P less than .05) in wethers fed sucrose and lactose, compared with those fed hay alone. All types of supplementary carbohydrates depressed (P less than .05) apparent absorption and urinary excretion of K. Serum Mg and Ca were not affected and serum K was depressed (P less than .05) by carbohydrate supplementation. Ruminal fluid pH was decreased (P less than .05) by glucose and lactose supplementation, and addition of these carbohydrates tended to decrease molar proportions of acetate and increase those of propionate and butyrate, compared with sheep fed hay alone. Sucrose addition decreased (P less than .05) acetate and increased (P less than .05) butyrate molar proportions in the ruminal fluid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pharmacokinetics of ivermectin after maternal or fetal intravenous administration in sheep

In pregnant sheep at 120–130 days of gestational age, a study was undertaken in order to characterize the pharmacokinetics and transplacental exchange of Ivermectin after maternal or fetal intravenous administration. Eight pregnant Suffolk Down sheep of 73.2 ± 3.7 kg body weight (bw) were surgically prepared in order to insert polyvinyl catheters in the fetal femoral artery and vein and amniotic sac. Following 48 h of recovery, the ewes were randomly assigned to two experimental groups. In group 1, (maternal injection) five ewes were treated with an intravenous bolus of 0.2 mg ivermectin/kg bw. In group 2, (fetal injection) three ewes were injected with an intravenous bolus of 1 mg of ivermectin to the fetus through a fetal femoral vein catheter. Maternal and fetal blood and amniotic fluid samples were taken before and after ivermectin administration for a period of 144 h post‐treatment. Samples were analyzed by liquid chromatography (HPLC). A computerized non‐compartmental pharmacokinetic analysis was performed and the results were compared by means of the Student t‐test. The main pharmacokinetic changes observed in the maternal compartment were increases in the volume of distribution and in the half‐life of elimination (t_½β). A limited maternal‐fetal transfer of ivermectin was evidenced by a low fetal Cmax (1.72 ± 0.6 ng/mL) and AUC (89.1 ± 11.4 ng·h/mL). While the fetal administration of ivermectin resulted in higher values of clearance (554.1 ± 177.9 mL/kg) and lower values of t_½β (8.0 ± 1.4 h) and mean residence time (8.0 ± 2.9 h) indicating that fetal‐placental unit is highly efficient in eliminating the drug as well as limiting the transfer of ivermectin from the maternal to fetal compartment.     

Transplacental exchange of moxidectin after maternal or fetal intravenous administration in sheep

The transplacental exchange of moxidectin after maternal or fetal intravenous (i.v.) administration was studied using the chronically catheterized fetal sheep model. Nine pregnant Suffolk Down sheep of 65.7 ± 5.9 kg body weight (bw) were surgically prepared to insert polyvinyl catheters in the fetal femoral artery and vein and amniotic sac. The ewes were randomly assigned to two experimental groups. In group 1 (maternal injection) five ewes were treated with an i.v. bolus of 0.2 mg of moxidectin/kg bw. In group 2, (fetal injection) an i.v. bolus of 1 mg of moxidectin was administered to the four fetuses by femoral vein catheters. Maternal and fetal blood and amniotic fluid samples were taken before and after moxidectin administration for a 144 h post-treatment period. Samples were analyzed by liquid chromatography. A noncompartmental pharmacokinetic analysis was performed and statistical differences were determined by mean of parametric and nonparametric statistical tests. Pharmacokinetic differences observed in maternal variables were shorter elimination half-life and mean residence time compared with values previously reported for ivermectin. Drug diffusion from maternal to fetal circulation ( AUC _{0– t}  = 232.6 ± 72.5 ng·h/mL) was statistically not different ( P =  0.09) compared with fetal to maternal diffusion ( AUC _{0– t} = 158.0 ± 21.6 ng·h/mL). Fetuses showed significantly ( P  =   0.008) lower drug body clearance values compared with those observed in the maternal side. Considering the observed transplacental passages between materno-fetal or feto-maternal circulations, we conclude that the placental barrier is not effective in preventing the moxidectin diffusion between mother and fetus.     

Effect of intraruminal butyrate infusion on the plasma insulin level in sheep

After intraruminal infusion of butyrate to sheep at dose rates of 0.25, 0.5, 1 and 2 g sodium n-butyrate per kg body mass, butyrate concentration of the rumen fluid and total secreted insulin rose in direct proportion to the butyrate dose infused. The half-life of butyrate in the rumen was always longer than that of insulin. At 90 min after the infusion of 1 g butyrate per kg body mass, butyrate concentration in the ruminal papillae reached the level corresponding to an extracellular concentration that reduced cell division by 50% in vitro. It can be concluded that butyrate may be present in the ruminal papillae in concentrations inhibiting cell proliferation, simultaneously with the presence of blood plasma insulin concentrations stimulating the proliferation of ruminal epithelial cells.     

The effects of maternal plasma dobutamine levels on fetal oxygenation in anaesthetized sheep

Objective To measure the effects of dobutamine infusion on fetal oxygenation during isoflurane anaesthesia in pregnant ewes. Study design Prospective randomized experimental study. Animals Seven clinically normal adult pregnant Rambouillet‐Dorset cross ewes with fetuses of 117–122 days gestational age. Methods The ewes were anaesthetized with ketamine (2 mg kg^?1) IM, and isoflurane (F_E′ISO 2.0%) in oxygen. After instrumentation and stabilization, dobutamine was infused at 4 µg kg^?1minute^?1 for 60 minutes and 10 µg kg^?1minute^?1 for 60 minutes in random order, separated by a 20‐minute washout period. Catheters were placed in the maternal and fetal carotid arteries; these were used for continuous blood pressure measurement and intermittent blood sampling. Results Maternal mean systemic carotid arterial pressure was 60 mm Hg prior to dobutamine infusion. After 5 minutes of dobutamine infusion, fetal oxygen saturation increased (p < 0.05) from 0.62 (0.17–0.71, minimum–maximum) to 0.72 (0.28–0.78) at a dose of 4 µg kg^?1minute^?1 and to 0.70 (0.20–0.73) at a dose of 10 µg kg^?1minute^?1. These increases were maintained during the infusion and were not significantly different between doses. Maternal oxygen saturation remained constant at 1.0 before and during all infusions. Although maternal heart rate and blood pressure increased (p < 0.05) by 90% and 25%, respectively, with dobutamine, this stimulant effect was not evident in the corresponding fetal variables. Maternal haemoglobin concentration increased 30% (p < 0.05) with each infusion. Conclusions Dobutamine at 4 µg kg^?1minute^?1 increases fetal oxygenation that is not improved by a dose of 10 µg kg^?1minute^?1. This increase is largely due to an increase in maternal haemoglobin concentration that, in turn, increases oxygen delivery to the placenta. Clinical relevance The use of dobutamine to treat hypotension in pregnant sheep during isoflurane anaesthesia improves fetal oxygenation. This may be true in other species.     

Evaluation of early fetal loss induced by gavage with eastern tent caterpillars in pregnant mares

OBJECTIVE: To determine whether gavage of pregnant mares (housed without access to pasture) with starved eastern tent caterpillars (ETCs) or their excreta is associated with early fetal loss (EFL), panophthalmitis, or pericarditis. DESIGN: Randomized clinical trial. ANIMALS: 15 mares. PROCEDURE: 15 mares with fetuses from 40 to 80 days of gestation (dGa) were randomly assigned to 1 of 3 groups and received 2.5 g of ETC excreta, 50 g of starved ETCs, or 500 mL of water, respectively, once daily for 10 days. Mares were housed in box stalls, walked twice daily, and not allowed access to pasture for 12 days before or during the 21-day trial. RESULTS: 4 of 5 mares gavaged with starved ETCs (group 2) aborted on trial days 8 (2 mares), 10, and 13. No control mares or mares that received excreta aborted. Differences between the ETC group and other groups were significant. Abortion occurred on 49, 64, 70, and 96 dGa. Allantoic fluids became hyperechoic the day before or the day of fetal death. Alpha streptococci were recovered from 1 fetus and Serratia marcescens from 3 fetuses. Neither panophthalmitis nor pericarditis was seen. The abortifacient component of the ETCs was not elucidated. CONCLUSIONS AND CLINICAL RELEVANCE: These findings suggest that mares with fetuses from 40 to 120 days of gestation should not be exposed to ETCs because they may induce abortion.     

Pharmacokinetics of chlortetracycline in maternal plasma and in fetal tissues following oral administration to pregnant ewes

The purpose of this study was to determine if concentrations of chlortetracycline could be detected in fetal plasma or tissues after administering an oral dose of chlortetracycline (CTC; 500 mg/head/day) reported to be effective in controlling Campylobacter spp. abortions. Five pregnant ewes were administered 250 mg/head twice a day (total dose 500 mg/hd/d) for 7 days. On the beginning of day 7, intravenous catheters were surgically implanted or inserted into the fetus and dam. Plasma samples were collected from the ewe and fetus at various time points before and up to 36 hr after the last dose of CTC. All ewes were then sacrificed, and tissues were harvested from the fetus for drug analysis. Concentrations of CTC in maternal plasma were consistent with our previous study and below the minimum inhibitory concentration of Campylobacter abortion isolates. Concentrations of CTC were below the limit of detection in three of five fetal plasma samples and all of the placenta, amniotic fluid, and fetal stomach contents. Low concentrations were detectable in fetal kidney and liver, suggesting that CTC reaches the fetus, although at a variable and low ratio when compared to maternal concentrations.     

The maternal to fetal transfer of immunoglobulins associated with placental lesions in sheep.

In this study we evaluated maternofetal transmission of immunoglobulins in ewes under conditions of altered placental morphology. Intravenous injection of human red blood cells was used to induce immunoglobulins in pregnant ewes. The hemagglutination test was used to detect antibody in maternal serum, fetal and placental fluids. Placental injury was induced by intravenous inoculation of Escherichia coli endotoxin or spores of Aspergillus fumigatus into pregnant ewes at days 99 or 100 of gestation respectively. Placental infarction, thrombosis of maternal placental vessels and variable neutrophil infiltrate characterized lesions produced by A. fumigatus. Endotoxin treated ewes developed marked placental edema, congestion, hemorrhage and focal loss of uterine epithelium. Human red blood cell agglutinating antibody was not detected in placental or fetal fluids obtained from ewes with either of the above placental lesions. Placentitis of undetermined etiology was observed in seven ewes. Two ewes had received A. fumigatus, two had received endotoxin and three were untreated ewes. Histological examination of their placentas revealed trophoblastic and endometrial epithelial necrosis and necrotizing vasculitis of the chorioallantois. Human red blood cell agglutinating antibody was detected only in the fetal and placental fluids of the seven ewes with these placental lesions. The nature of these lesions would have produced a functional confluence of the maternal and fetal circulations. Antibody transfer from dam to fetus was observed only in association with placental lesions which produced this confluence of circulations. The character of the placental lesions, rather than the mere presence of placental lesions apparently determined the transfer of immunoglobulins.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of isulin-like growth factor-I on maternal and fetal plasma amino acid levels in pregnant rats

Pregnant rats were subcutaneously administered with recombinant human insulin-like growth factor-I (rhIGF-I) in doses of 0 (control), 1, 2, and 4 microg/g body weight per day from day 18 to 21 of pregnancy. On day 21 of pregnancy, maternal and fetal plasma samples were collected and those amino acid levels were measured. The ratios of fetal/maternal plasma amino acids, especially leucine, isoleucine, tryptophan, phenylalanine and tyrosine, increased in 2 microg rhIGF-I treated group. Both total fetal weight and total placental weight were also higher than those in the control group. These results suggested that IGF-I enhanced fetal growth by, as one of its possible mechanisms, promoting placental amino acid supplies from the mother to fetuses.     

Effect of short term intravenous infusion of noradrenaline on energy metabolism in shorn and unshorn pregnant sheep

The effect of a three hour intravenous noradrenaline (NA) infusion, at a dose of 0.8 micrograms min-1 kg-1 bodyweight, on energy metabolism was studied in five pairs of shorn and unshorn pregnant ewes. The concentration of oxygen and carbon dioxide in blood was significantly higher in shorn animals during saline infusion, but this difference between shorn and unshorn groups was removed by NA infusion. Significant increases in the plasma concentrations of glucose, lactate, non-esterified fatty acids (NEFA), 3-hydroxybutyrate and glycerol occurred in both shorn and unshorn animals during NA infusion. There was a 48 per cent larger rise in the plasma NEFA concentration during NA infusion in the unshorn group, suggesting that the lipolytic effect of exogenous NA is greater in these animals. NA infusion had no effect on the plasma concentration of thyroid hormones in both groups of ewes, while growth hormone levels fell and cortisol increased in all animals. The plasma insulin concentration was significantly reduced during NA treatment in the unshorn group, but was unchanged in shorn animals. These results may be a consequence of sympathetic activity already being elevated in shorn ewes due to chronic exposure to an environmental temperature below the thermoneutral zone.     

Distribution of lesions in fetal brains following experimental infection of pregnant sheep with Toxoplasma gondii

Six ovine fetal brains were harvested 33 to 35 days postchallenge from 5 ewes, each of which was given 3000 Toxoplasma gondii oocysts on day 90 of pregnancy. Histopathologic examination of transverse sections taken at 13 levels in the fetal brains revealed the presence of toxoplasmosis-related lesions in all 6 brains. However, lesions were not randomly distributed (P = .007); they were most numerous at the level of the optic tract, the rostral margin of the pons, and 4 mm caudal to the ansate sulcus and were absent in all sections at the level of the caudal cerebellum. Lesion distribution may be due to hemodynamic factors, differences in the expression of endothelial surface receptor molecules at the level of the blood-brain barrier, or the presence of localized permissive/inhibitory factors within the brain. The results have implications for the selection of areas of brain from aborted ovine fetuses to be examined histopathologically for laboratory diagnosis.     

Effects of selenium supply and dietary restriction on maternal and fetal metabolic hormones in pregnant ewe lambs

The objective of these studies was to evaluate the effects of dietary restriction and Se on maternal and fetal metabolic hormones. In Exp. 1, pregnant ewe lambs (n = 32; BW = 45.6 +/- 2.3 kg) were allotted randomly to 1 of 4 treatments. Diets contained (DM basis) either no added Se (control), or supranutritional Se added as high-Se wheat at 3.0 mg/kg (Se-wheat), or sodium selenate at 3 (Se3) and 15 (Se15) mg/kg of Se. Diets (DM basis) were similar in CP (15.5%) and ME (2.68 Mcal/kg). Treatments were initiated at 50 +/- 5 d of gestation. The control, Se-wheat, Se3, and Se15 treatments provided 2.5, 75, 75, and 375 microg/kg of BW of Se, respectively. Ewe jugular blood samples were collected at 50, 64, 78, 92, 106, 120, and 134 d of gestation. Fetal serum samples were collected at necropsy on d 134. In Exp. 2, pregnant ewe lambs (n = 36; BW 53.8 +/- 1.3 kg) were allotted randomly to treatments in a 2 x 2 factorial arrangement. Factors were nutrition (control, 100% of requirements vs. restricted nutrition, 60% of control) and dietary Se (adequate Se, 6 microg/kg of BW vs. high Se, 80 microg/kg of BW). Selenium treatments were initiated 21 d before breeding, and nutritional treatments were initiated on d 64 of gestation. Diets were 16% CP and 2.12 Mcal/kg of ME (DM basis). Blood samples were collected from the ewes at 62, 76, 90, 104, 118, 132, and 135 d of gestation. Fetal blood was collected at necropsy on d 135. In Exp.1, dietary Se source and concentration had no effect (P > 0.17) on maternal and fetal serum IGF-I, triiodothyronine (T(3)), or thyroxine (T(4)) concentrations. Selenium supplementation increased (P = 0.06) the T(4):T(3) ratio vs. controls. In Exp. 2, dietary Se had no impact (P > 0.33) on main effect means for maternal and fetal serum IGF-I, T(3), or T(4) concentrations from d 62 to 132; however, at d 135, high-Se ewes had lower (P = 0.01) serum T(4) concentrations than adequate-Se ewes. A nutrition by Se interaction (P = 0.06) was detected for the T(4):T(3) ratios; ewes fed restricted and adequate-Se diets had greater (P = 0.10) T(4):T(3) ratios compared with the other treatments. Nutrient-restricted ewes had lower (P < 0.05) serum IGF-I, T(3), and T(4) concentrations. Fetal serum IGF-I concentrations were lower (P = 0.01) in restricted-vs. control-fed ewes; however, fetal T(3) and T(4) concentrations were unaffected (P > 0.13) by dietary Se or maternal plane of nutrition. These data indicate that dietary Se may alter maternal T(4):T(3) ratios. In addition, nutrient restriction during gestation reduces maternal IGF-I, T(3), and T(4) and fetal IGF-I concentrations.     

Differences in signs and lesions in sheep and goats with enterotoxemia induced by intraduodenal infusion of Clostridium perfringens type D

Enterotoxemia was induced in 4 lambs and 4 goat kids by continuous intraduodenal infusion of a whole culture of Clostridium perfringens type D. Clinical signs, hematologic values, biochemical alterations, and postmortem lesions in the lambs and goat kids were compared. The 4 lambs and 4 goat kids died within 25 hours of beginning the infusions. Lesions were not observed in the gastrointestinal tract of the 4 lambs; however, severe hemorrhagic enterocolitis was found in the 4 goat kids. This difference between the lambs and goat kids in the lesions caused by experimentally induced enterotoxemia may explain the discrepancies reported between sheep and goats in clinical signs, response to treatment, and efficacy of vaccination observed in naturally induced enterotoxemia in the 2 species.     

Brucellosis induced by RB51 vaccine in a pregnant heifer

Brucellosis developed in a 14.5-month-old Gelbvieh heifer after the animal was vaccinated with the calfhood dose of strain RB51 Brucella abortus vaccine s.c. during the fourth month of its first pregnancy. The heifer experienced dystocia and was euthanatized during cesarean section because of a large uterine tear. The fetus was dead at delivery. Suppurative placentitis and fetal pneumonia were evident at necropsy. Brucella abortus strain RB51 was isolated from the placenta and the fetus' lung.