The effect of prednisolone on canine neutrophil function: in vivo and in vitro studies.

The in vivo effect of a therapeutic dose of prednisolone on canine neutrophil adherence, random migration, chemotaxis, phagocytosis of IgG and C3b opsonized yeast cells, chemiluminescence, Fc- and CR3-receptor expression was investigated. Prednisolone was also added in vitro to neutrophils as isolated cells and in whole blood. In the in vivo study, prednisolone increased the IgG mediated ingestion of yeast cells and the number of activated neutrophils in the phagocytosis assay, while flow cytometric investigation of the IgG-receptor Fc gamma RIII with a monoclonal antibody showed similar expression before, during and after treatment. Prednisolone also increased the ingestion of C3b-opsonized yeast cells, while the expression of CR3-receptors (CD11b CD18) measured by flow cytometry was unchanged. Chemiluminescence and the chemotactic response towards zymosan activated serum were increased, while adherence to nylon wool was decreased. The in vitro studies revealed that prednisolone had no or a dampening effect on neutrophils in cell suspensions. Adherence as well as IgG mediated ingestion was decreased at the highest prednisolone concentration (800 ng/ml) in whole blood. The present study suggests that the part of the antiinflammatory effect of corticosteroids mediated through their influence on neutrophils, besides reduced adherence, may be exerted by increased clearance of microorganisms and IgG-complexes through an elevated functional capacity.

Plasma concentrations of 15-ketodihydro-PGF2 alpha, progesterone, oestrone sulphate, oestradiol-17 beta and cortisol during late gestation, parturition and the early post partum period in llamas and alpacas.

Plasma concentrations of 15-ketodihydroprostaglandin (PG) F2 alpha, progesterone, oestrone sulphate, oestradiol-17 beta and cortisol during late gestation, parturition and the early post-partum period were measured in six llamas and five alpacas. During the last 100 days of pregnancy, 15-ketodihydro-PGF2 alpha concentrations increased steadily until the day of parturition when a massive release was detected (P < 0.01) concomitant with a decrease in progesterone concentrations (P < 0.01). The highest PGF2 alpha metabolite concentrations (159 +/- 35 nmol l-1 and 92 +/- 29 nmol l-1 in llamas and alpacas respectively) were detected in the sample collected during the morning on the day of parturition. Basal concentrations were registered by day 3 after delivery. Plasma concentrations of oestrone sulphate started to increase 80 days before parturition and reached peak concentrations immediately before parturition (15 +/- 3 nmol l-1 in llamas and 18 +/- 5 nmol l-1 in alpacas). Oestrone sulphate concentrations dropped sharply (P < 0.01) on the day of parturition in llamas and one day later in alpacas, whereupon they remained relatively unchanged until at least 20 days postpartum. Oestradiol-17 beta concentrations were higher than 180 pmol l-1 during the last 45 days of pregnancy, began to decrease on the day of parturition and reached very low concentrations within the following two days. High oestradiol-17 beta concentrations were registered 7 days postpartum in all alpacas (P < 0.05) and within 10 days of parturition in five of six llamas (P < 0.01). No significant cortisol peaks were observed around parturition, but mean concentrations were increased in both species.

Plasma concentrations of progesterone, oestradiol, LH and 15-ketodihydro-PGF2 alpha in Norwegian semi-domestic reindeer (Rangifer tarandus tarandus) during their first reproductive season.

Blood samples were collected from eleven 1.5 year old female reindeer three times a week from September 1992 until February 1993 and daily in October and November 1992. Blood samples were collected every third hour for a period of three weeks from two females that showed regular oestrous cycles. Plasma progesterone, oestradiol, LH and 15-ketodihydro-PGF2 alpha were analysed to characterize variations in ovarian function. Reindeer are seasonally polyoestrous. Average duration of oestrous cycles was 19.4 days (range: 13-33 days). Short periods of high progesterone concentrations (4-8 days duration) occurred in five animals before the onset of regular oestrous cycles. The first regular cycle had a significantly longer luteal phase than did the following cycles. During luteolysis 15-ketodihydro-PGF2 alpha was released in a pulsatile pattern. Maximal oestradiol concentrations preceded preovulatory LH peaks by about 3 h in the two frequently sampled animals. The duration of LH surges was 12 and 15 h, respectively.

Effects of calf removal at parturition on postpartum ovarian activity in Zebu (Bos indicus) cows in the humid tropics.

To assess endocrine and morphological responses of ovaries to total weaning at parturition, 6 Zebu (Bos indicus) cows 5 years or older were investigated. Following parturition, blood samples were collected daily during the first month and twice weekly thereafter until day 60 to determine concentrations of progesterone (P4) and prostaglandin F2 alpha metabolite. It took between 25 to 32 days to complete uterine involution. The prostaglandin metabolite remained elevated for a mean period of 14.2 days (range, 4-21) postpartum. Five of the animals resumed cyclicity with a short estrous cycle starting between days 7 to 34 and lasting between 7 and 14 days. No estrous behavior was recorded prior to the short estrous cycles, but subsequent normal-length estrous cycles were all preceded by signs of estrus. In the 1 animal that resumed cyclicity with an estrous cycle of normal length on day 37 (length 20 days), the cycle was preceded by estrous behavior. Progesterone concentrations reached a mean maximum of 4.8 nmol liter-1 during the short estrous cycles, and prostaglandin metabolite concentrations peaked while P4 concentrations were decreasing. P4 concentrations reached a mean maximum of 12.2 nmol liter-1 during the estrous cycles of normal length. The interval from parturition to the first estrous cycle of normal length varied between 16 and 48 days, and the length of the cycle was 18 to 22 days. Starting 2 days postpartum, ovaries from 5 of the cows were scanned by ultrasonography every second day until day 30 postpartum. Medium-sized follicles were detected between days 4 to 7 postpartum in 4 of the scanned cows that later had short estrous cycles. The time between parturition and the appearance of the first dominant follicle was 7.6 days (range 6-10 days). The interval between parturition and the appearance of the first ovulatory-sized follicle was 10.2 days (range 8-13 days). In 3 of the scanned cows this ovulatory-sized follicle ovulated. We conclude that cyclic ovarian activity in Zebu cows can start early in the postpartum period in the absence of offspring, and that short luteal phases, not preceded by estrous behavior, may play an important role in establishing normal postpartum ovarian activity.

Endocrine changes after mating in pregnant and non-pregnant llamas and alpacas.

Plasma concentrations of oestradiol-17 beta, progesterone, 15-keto-dihydro-PGF2 alpha and luteinizing hormone (LH) were monitored in llamas and alpacas after mating with an intact male. Concentrations of LH and PGF2 alpha metabolite were high immediately after copulation. Ovulation occurred in 92% of the animals. The first significant increases in progesterone were recorded on day 4 after mating. In non-pregnant animals the lifespan of the corpus luteum was estimated to be 8-9 days. Luteolysis occurred in association with the release of PGF2 alpha. In pregnant animals, a transient decrease in progesterone concentrations was observed between days 8 and 18 in both species. No significant changes in PGF2 alpha secretion were registered during this period. Oestradiol-17 beta concentrations were high on the day of mating, declined to low values on day 4, and started to increase again on day 8. Peak values after luteolysis in non-pregnant animals were significantly higher than those registered in pregnant ones. Furthermore, concentrations of oestradiol-17 beta were elevated for a longer period in non-pregnant than in pregnant animals. The results suggest that progesterone from the corpus luteum exerts a negative influence on follicular activity in pregnant animals by reducing oestradiol-17 beta secretion.

Hematological, blood biochemical, and cytological bronchoalveolar lavage studies in prepubertal gilts after endotoxin inhalation and ingestion.

The effect of oral intake of endotoxin (ET) combined with inhalation of ET-contaminated feed-dust was studied in 6 prepubertal gilts. The animals were given 100 mg of ET each in their ordinary feed ration and meanwhile stayed for 4 h in a continuous flow exposure chamber where the endotoxin concentration was approximately 9 micrograms/m3. Blood samples were collected during the exposure and for 8 h afterwards. Bronchoalveolar lavage (BAL) was performed 24-26 h after the exposure. Six additional animals received ordinary feed and were exposed to feed-dust without added ET. Blood samples were analysed for total white blood cell count (B-WBC) and differential count: polymorphonuclear (PMN) and mononuclear (MN); total red blood cell count (B-RBC); platelets (B-PLT); total serum bile acids (S-BA); glutamate dehydrogenase (S-GLDH); and a blood plasma prostaglandin F2 alpha metabolite (15-ketodihydro-PGF2 alpha; P-PG). The major findings were decreased numbers of MN, PMN and B-RBC 3 to 6 h after the start of the ET exposure and increased numbers of neutrophilic granulocytes in the BAL fluid of the ET-exposed animals. It is concluded that a combined oral and airborne ET challenge induces hematological and cytological changes that may contribute to the development of respiratory disorders.

Effect of induced suprabasal progesterone levels around estrus on plasma concentrations of progesterone, estradiol-17beta and LH in heifers.

A controlled study was carried out to investigate the effects of suprabasal plasma progesterone concentrations on blood plasma patterns of progesterone, LH and estradiol-17beta around estrus. Heifers were assigned to receive subcutaneous silicone implants containing 2.5 g (n=4), 5 g (n=4), 6 g (n=3), 7.5 g (n=3) or 10 g (n=4) of progesterone, or implants without hormone (controls, n=5). The implants were inserted on Day 8 of the cycle (Day 0=ovulation) and left in place for 17 d. The time of ovulation was determined by ultrasound scanning. Blood was collected daily from Days 0 to 14 and at 2 to 4-h intervals from Days 15 to 27. Control heifers had the lowest progesterone concentrations on Days 20.5 to 21 (0.5 +/- 0.1 nmol L(-1)); a similar pattern was observed in heifers treated with 2.5 and 5 g of progesterone. In the same period, mean progesterone concentrations in the heifers treated with 6, 7.5 and 10 g were larger (P < 0.05) than in the controls, remaining between 1 and 2.4 nmol L(-1) until implant removal. A preovulatory estradiol increase started on Days 16.4 to 18.4 in all the animals. In the controls and in heifers treated with 2.5 and 5 g of progesterone, estradiol peaked and was followed by the onset of an LH surge. In the remaining treatments, estradiol release was prolonged and increased (P < 0.05), while the LH peak was delayed (P < 0.05) until the end of the increase in estradiol concentration. The estrous cycle was consequently extended (P < 0.05). In all heifers, onset of the LH surge occurred when progesterone reached 0.4 to 1.2 nmol L(-1). The induction of suprabasal levels of progesterone after spontaneous luteolysis caused endocrine asynchronies similar to those observed in cases of repeat breeding. It is suggested that suprabasal concentrations of progesterone around estrus may be a cause of disturbances oestrus/ovulation.

Body weight loss during lactation in relation to energy and protein metabolism in standard-fed primiparous sows.

After 5 weeks of lactation 14 standard-fed primiparous sows were divided into a low weight-loss group (L-gr, loss < 25 kg, n = 7) and a high weight-loss group (H-gr, loss > or = 25 kg, n = 7). Body weights of the sows and their litters were recorded on days 2, 7, 14, 21, 28 and 35 of lactation. Blood samples were collected before the morning feeding on each weighing day. Samples were analysed to determine concentrations of insulin, glucagon, glucose, triglycerides, non-esterified fatty acids (NEFA), urea and creatinine. The H-gr sows lost weight throughout lactation, whereas the L-gr sows gained weight during the last week. Weight loss was higher in the H-gr than in the L-gr during weeks 2, 3 and 5 of lactation. Litter size and litter weight gain were higher in the H-gr than in the L-gr. Significant changes in levels of insulin, glucagon, glucose, triglycerides and creatinine were observed over lactation. No significant differences in concentrations of any of the parameters were found between the two groups, except for insulin and cholesterol which were higher in the L-gr. Catabolism of adipose tissue and muscle protein was observed in both groups during the first week of lactation. This catabolic state was more pronounced and tended to be prolonged in the H-gr. Concentrations of all parameters seemed to be stable in both groups during the last two weeks of lactation.

Effects of insulin administration at weaning on hormonal patterns and reproductive performance in primiparous sows.

Twelve primiparous sows were grouped into six pairs. One sow from each pair (T) received subcutaneous injections of porcine insulin as follows: pairs 1 and 2, 0.5 IU/kg at 07.00 h; pairs 3 to 6, 0.5 IU/kg at 07.00 and 15.00 h. Insulin administration was started one day before weaning and continued until the last day of post-weaning oestrus. Control sows (C) received physiological saline solution as a sham treatment. Blood samples were collected every 3 h from 09.00 to 21.00 h and, additionally, at 07.00 h during the period of insulin treatment. Additional blood samples were collected at 15-min intervals for 6 h for 2 days before and for 3 days after weaning, and for 12 h on the day of weaning (D35 of lactation). Exogenous treatment with insulin increased plasma levels of insulin (53 v 36 mU/l, P < 0.001) and decreased plasma levels of glucose (4.6 v 5.5 mmol/l, P < 0.001) in T-sows as compared to C-sows. During the treatment period, the average levels of cortisol (31 v 35 nmol/l, P < 0.001) and non-esterified fatty acids (0.13 v 0.16 mmol/l, P < 0.01) were lower in T-than in C-sows. No differences in glucagon or urea levels were observed between the two groups. Plasma levels of LH were higher in T- than in C-sows both before and during the treatment period. However, no differences in number of LH pulses/6h were found between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of oral and intravenous administration of endotoxin in prepubertal gilts.

The effect of oral intake of endotoxins was studied in 12 prepubertal gilts. The animals were given 30 or 100 mg of ET each in their regular morning feed ration. Blood samples were collected periodically during 24 h and the clinical status, including rectal temperature, was recorded at the same time. Hematological and clinical chemical analyses that included serum bile acids, glutamate dehydrogenase, alkaline phosphatase, calcium, iron, zinc and a blood plasma metabolite of prostaglandin F2 alpha, were done. The animals showed no obvious clinical symptoms following endotoxin feeding. The major findings were increased bile acid and glutamate dehydrogenase values with the most prominent rises being recorded 10-12 h after endotoxin intake. In a later experiment, 6 animals were injected i.v. with endotoxin in doses in the range 0.1-0.5 micrograms/kg b.w. Blood samples were taken and analysed as in the endotoxin-feeding experiment. Within 1 h of injection, all animals showed symptoms such as vomiting, fever and dyspnea. The clinical signs disappeared within 2-5 h. The injections were followed by increases in bile acids, glutamate dehydrogenase and prostaglandin F2 alpha metabolite. To conclude, this study indicates that clinically healthy prepubertal gilts react to ingested endotoxin in feed but that no apparent clinical disturbances ensue.

Reduced response to intravenous endotoxin injections following repeated oral administration of endotoxin in the pig.

Three prepubertal gilts were each given 100 mg of endotoxin (ET) in their ordinary feed rations, twice daily for 6 days; 3 other gilts received standard feed. Following ET feeding, all animals were injected intravenously (i.v.) with ET (1.0 microgram/kg b.w.) once daily for 5 days. Blood samples were collected and analysed for hematology and total serum bile acids (S-BA), glutamate dehydrogenase (S-GLDH), calcium (S-Ca), iron (S-Fe), zinc (S-Zn) and a blood plasma metabolite (15-ketodihydro-PGF2a; P-PG) of prostaglandin F2a. The animals showed no apparent clinical symptoms following ET-feeding, neither did the blood analyses reveal effects of oral ET. However, when iv ET injections were given, the ET-fed animals showed fewer clinical signs of endotoxemia following the 2nd to 5th injection. S-BA and S-GLDH increased markedly in the standard-fed group following the first injection, while the ET-fed animals showed a much smaller increase in S-BA and no change in S-GLDH on that day. The difference in response may be explained by a direct uptake of ET from the gastrointestinal tract in the ET-fed pigs, making them less sensitive to the injected ET.

Relationship between lactation-associated body weight loss, levels of metabolic and reproductive hormones and weaning-to-oestrous interval in primiparous sows.

Twelve conventionally fed primiparous sows with a similar number of piglets were divided into 3 groups (n = 4) according to their body weight loss after a 5-week lactation as follows: H-gr (loss greater than 25 kg), M-gr (loss 11-25 kg) and L-gr (loss less than or equal to 10 kg). Blood samples were collected at 9, 12, 15, 18 and 21 h, from 4 days before weaning until 3 days afterwards, and every 15 min for 6 h on the day before and the day after weaning. The group of sows with higher weight loss during lactation tended to have a higher farrowing weight and higher pre-weaning plasma prolactin levels. At weaning, the L-gr sows had higher plasma insulin and lower plasma cortisol concentrations, especially when compared with H-gr sows. Among the ten sows that showed oestrus within 10 days of weaning, no between-group differences were found in the length of the interval from weaning to oestrus. No significant differences between groups were observed in levels of oestradiol-17 beta and LH. The higher level of insulin in L-gr sows might relate to the higher number of corpora lutea observed in this group.

Factors affecting skim milk progesterone assay results.

Five studies were performed to determine factors affecting progesterone concentration in skim milk. Results of the first study indicated that progesterone concentration was higher in skim milk of samples kept 16 hours in an ice bath (0 C) than of those left at room temperature (21 C). In the second study, this temperature effect was found to be reversible, with skim milk progesterone concentration increasing when whole milk samples were cooled prior to centrifugation. In the third study, [3H]-labeled progesterone was used to determine the relationship between fat content of foremilk (the first milk obtained from the teats), midmilk (milk obtained midway through milking), and strippings (milk obtained immediately after milking machines have been removed) samples and temperature (4 C and 21 C) on the percentage of progesterone in the skim milk fraction. The relationship between percentage of butterfat and percentage of progesterone in skim milk was linear when the log of these variables was used for calculations. In the fourth study, assayable progesterone in the skim milk fraction of foremilk, midmilk, and strippings was affected by temperature. In the fifth study, a multiple-regression procedure was used to determine the amount of variation in percentage of radioactive progesterone in the skim milk fraction. Independent variables (whole milk butterfat and temperature of incubation [1, 3, 13, 22, 37, and 50 C]) and the natural log of each variable, were entered into a stepwise multiple-regression analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Testicular ultrastructure of Zebu bulls in Costa Rica.

The testicular ultrastructure of sexually mature Zebu (Bos indicus) bulls from Costa Rica was studied in relation to their classification based on andrological examinations undertaken in the field. Based on the testicular consistency at palpation, each bull was assigned to one of three groups, ranging from normal (Group A) to markedly reduced testicular consistency (Group C). The ultrastructure of the clinically normal testicle (Group A) in the Zebu bull resembled that of Bos taurus. In testicles with a slight to moderate reduction of testicular consistency (Group B) degenerated primary spermatocytes and spermatids were common at all stages of the seminiferous cycle. Abnormalities in the condensation of the chromatin, acrosomal defects, and the presence of degenerated cells and cellular debris were the most common of disturbances found. In testicles from bulls with a marked reduction of testicular consistency (Group C) the seminiferous epithelium of most tubules showed degenerative changes, thereby making it impossible to classify the stage of the cycle. The changes were mainly restricted to spermatocytes and spermatids, whose relative numbers were reduced. The number of cells undergoing meiotic divisions was also greatly reduced. Changes in the spermatids were also found including the appearance of severe acrosomal defects, such as acrosomal pouches, the failure of nuclear elongation and clear defects in chromatin condensation. The present ultrastructural findings in Bos indicus do not differ from the picture of normal testicular structure and of testicular degeneration usually found in Bos taurus bulls.

Cortisol binding capacity of corticosteroid binding globulin in hyperadrenocorticoid and healthy dogs.

Corticosteroid binding globulin (CBG) binding capacity and plasma cortisol concentration were determined in 27 dogs with hyperadrenocorticism and in 17 healthy control dogs. Cortisol concentrations were significantly higher in hyperadrenocorticoid dogs than in controls. CBG binding capacity did not differ between the two groups. It is concluded that excessive endogenous cortisol secretion does not induce an increase in CBG binding capacity. Consequently, the determination of CBG binding capacity provides no additional information for the diagnosis of canine Cushing's syndrome.

A possible role of endotoxins in spontaneous paretic cows around parturition.

Six late pregnant old cows which had a history of parturient paresis were studied with respect to clinical and blood biochemical changes during two weeks before and after parturition. The levels of serum endotoxin were evaluated during two days before and after parturition. Three out of six animals showed signs of paresis and two animals needed treatment. The clinical and general changes of all parameters supported the idea that during parturient paresis and parturition, there is absorption of endotoxins from the gastro-intestinal tract. Most of the studied parameters changed similarly to what has been described in experimentally induced endotoxaemia. The animals showed signs of ruminal stasis, had an increase in body temperature, prostaglandin metabolite and endotoxin as well as a decrease in serum calcium, zinc, iron and bile acids levels. The changes of these parameters were also closely related to each other. Furthermore, the changes were more pronounced in paretic cows. Endotoxins absorbed from the gastro-intestinal tract can be involved in paresis and make the outcome of this disease more severe.

Hormonal interrelationships in postpartum suckled dairy cows.

Three cross-bred cows calved in March and April and were followed until day 62 after parturition. Each animal was suckled by 2 calves ad libitum. All calves were removed from the cows on day 55 after parturition. Blood was collected 3 times per day from the jugular vein by venipuncture. On 4 occasions after parturition--i.e. days 7-8, 21-22, 35-36 and 49-50, the cows were bled through a jugular venous catheter every 30 min during the 24 h. The plasma samples were analyzed for the content of 15-keto-13,14-dihydro-PGF2 alpha (main PGF2 alpha metabolite), LH, prolactin, cortisol and progesterone by radioimmunoassay methods. The concentration of PGF2 alpha increased from 280 to 730 pmol/l within the last 4 days before parturition. The highest geometric mean was 3106 pmol/l on the day of parturition. Thereafter a steady decrease of PGF2 alpha metabolite concentration was seen until day 21 when it reached plateau at 148 pmol/l. In all cows plasma LH concentrations increased significantly (P < 0.05) from about 1.6 micrograms/l on days 7-8 to 2.4 micrograms/l on days 21-22 post partum. The frequency of LH pulses showed no tendency to increase as the postpartum period progressed and averaged 6.5 pulses/24 h. Mean plasma LH concentrations increased from 2.1 micrograms/l 2 days before weaning to 3.2 micrograms/l 2 days after weaning (P < 0.05). LH peaks occurred less frequently in association with prolactin and cortisol peaks than in their absence. A partial positive correlation between PGF2 alpha metabolite and cortisol (r = 0.30) was found on days 7-8 post partum. Correlation between prolactin and cortisol on days 7-8 and 21-22 post partum was also positive (r = 0.20 and r = 0.27, respectively). There was a negative correlation between LH and cortisol on days 7-8 (r = -0.27) and days 49-50 (r = -0.21) post partum. The first and short progesterone increase observed after weaning was terminated in conjunction with PGF2 alpha metabolite peaks.

The role of endotoxins in induced ruminal acidosis in calves.

Experimentally induced ruminal acidosis was carried out in 4 calves to investigate the possibility of resorption of endotoxins from Gram negative bacteria originating from the gastro-intestinal tract. The ruminal acidosis was induced by overfeeding of oats and the effect was evaluated by clinical and blood biochemical changes. Blood samples were collected every 2 h for 60 h before and after experimental feeding. The animals showed signs of ruminal acidosis and also the clinical and blood biochemical changes were similar to those seen after experimental endotoxaemia. However, although being less dramatic the changes in many relevant parameters such as an increase in prostaglandin F metabolite levels, body temperature, endotoxins and a decrease in iron indicate that an endotoxaemic state had occurred. The results of the present study show that ruminal acidosis/stasis is linked to resorption of endotoxins from the gastro-intestinal tract causing endotoxaemia/endotoxicosis.

Progesterone and oestradiol-17 beta receptors in the porcine myometrium during the oestrous cycle.

The concentrations of the tissue receptors for oestradiol (E) and progesterone (P) in the porcine myometrium at different stages of the oestrous cycle have been investigated by in vitro binding and exchange methods. Both hormones bound to specific cytoplasmic (Rc) and nuclear (Rn) receptors proteins with high affinity. The amount of ERc did not vary significantly throughout the cycle. Significant variations in the amount of myometrial ERn were observed with high concentrations during late pro-oestrus. The amount of PRc reached a top value in early pro-oestrus. A significant increase in PRn levels was observed at early luteal phase, and it remained high during the rest of the luteal phase. The temporal relationships between the variations in levels of oestradiol and progesterone nuclear receptors in myometrial tissue and those of the circulating plasma levels were established. The data obtained in this study suggest a relationship between the changes in levels of oestradiol and progesterone myometrial binding and the uterine motility pattern during the oestrous cycle.