Canine Pancreatic‐Specific Lipase Concentrations in Clinically Healthy Dogs and Dogs with Naturally Occurring Hyperadrenocorticism

Background

Specificity of canine pancreatic lipase immunoreactivity (cPLI) assays in dogs with hyperadrenocorticism (HAC) is unknown.

Hypothesis

Results of cPLI assays differ for clinically healthy dogs and dogs with HAC.

Animals

Seventeen healthy dogs and 20 dogs with HAC diagnosed by ACTH stimulation test results without evidence of clinical pancreatitis.

Methods

Dogs were enrolled between December 2009 and November 2010. Serum cPLI concentrations were determined by quantitative (Spec cPL test, SPEC) and semiquantitative (SNAP cPL test, SNAP) assays. Results were categorized as normal, equivocal, or abnormal (SPEC) or negative or positive (SNAP). Associations between group and cPLI were assessed using Fisher''s exact test or the Mann–Whitney U‐test. Spearman rank correlation coefficients (ρ) were determined for SNAP and SPEC results. Significance was set at P < .05.

Results

Spec cPL test concentrations were significantly (P < .001) higher in dogs with HAC (491.1 μg/L) than in healthy dogs (75.2 μg/L), with more abnormal SPEC results in HAC dogs (P < .001). There were more (P = .002) positive SNAP results in dogs with HAC (55%) than in healthy dogs (6%). SNAP and SPEC results were highly correlated (ρ = 0.85; P < .001).

Conclusions and Clinical Importance

Dogs with HAC had higher SPEC concentrations and more positive SNAP results than clinically healthy dogs with normal ACTH stimulation test results. Specificity of SPEC and SNAP assays in HAC dogs without clinical pancreatitis were 65 and 45%, respectively. Pending further study, SNAP and SPEC results should be interpreted cautiously in dogs with HAC to avoid false diagnosis of concurrent pancreatitis.     

Physiological indicators of stress and meat and carcass characteristics in tail bitten slaughter pigs

Background

Tail biting is a common welfare problem in pig production and in addition to being a sign of underlying welfare problems, tail biting reduces welfare in itself. The aim of this study was to evaluate the effects of tail biting on different pre and post mortem indicators of stress in slaughter pigs and on carcass and meat characteristics. A total of 12 tail bitten (TB) and 13 control (C) pigs from a farm with a long-term tail biting problem were selected for salivary cortisol analyses before and after transport to the slaughterhouse. After stunning, samples were taken for the analysis of serum cortisol, blood lactate, intestinal heat shock protein 70 (HSP70), and meat quality characteristics. In addition, body temperature immediately after and muscle temperature 35 min after stunning were measured, as well as lean meat percentage and carcass weight.

Results

TB pigs showed a lower cortisol response to the transport-induced stress than C pigs and also had a lower serum cortisol concentration after stunning. HSP70 content in the small intestine was higher in the TB pigs than in C pigs. TB pigs had a considerably lower carcass weight therefore produced a lower total amount of lean meat per carcass than C pigs.

Conclusions

This study suggests that prolonged or repeated stress in the form of tail biting causes a blunted stress response, possibly a sign of hypocortisolism. In addition, it underlines the importance of reducing tail biting, both from an animal welfare and an economic point-of-view.     

Ovarian localization of 11β-hydroxysteroid dehydrogenase (11βHSD): effects of ACTH stimulation and its relationship with bovine cystic ovarian disease

Cystic ovarian disease (COD) is an important cause of infertility in cattle, and ACTH has been involved in regulatory mechanisms related to ovarian function associated with ovulation, steroidogenesis, and luteal function. Here, we examined the localization of 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) and 11βHSD2 proteins in the ovary of healthy cows and animals with spontaneous and ACTH-induced COD and the in vitro response of the follicular wall exposed to ACTH. After stimulation by ACTH, we documented changes in 11βHSD expression and cortisol secretion by the follicular wall of large antral and follicular cysts. Follicular cysts showed a higher constitutive expression of both enzymes, whereas ACTH induced an increase in 11βHSD1 in tertiary follicles and follicular cysts and a decrease in 11βHSD2 in follicular cysts. Moderate expression of 11βHSD1 was observed by immunohistochemistry in granulosa of control animals, with an increase (P < 0.05) from primary to secondary, tertiary, and atretic follicles. The level of immunostaining in theca interna was lower than that in granulosa. The expression of 11βHSD2 was lower in the granulosa of primary follicles than in that of secondary, tertiary, and atretic follicles and was lower in the theca interna than in the granulosa. In ACTH-induced and spontaneously occurring follicular cysts, differences from controls were observed only in the expression of 11βHSD1 in the granulosa, being higher (P < 0.05) than in tertiary follicles. These findings indicate that follicular cysts may be exposed to high local concentrations of active glucocorticoids and indicate a local role for cortisol in COD pathogenesis and in regulatory mechanisms of ovarian function.     

Effect of Changes in Light on Hormonal Secretion and Milk Production of Dairy Cows in Early Lactation

Abstract

Two experiments were carried out to investigate the effect of a change in day length and light intensity on the concentration of prolactin in milk and plasma, on milk production during the first two months of lactation, and on the daily variation in plasma concentrations of prolactin, melatonin, growth hormone, Cortisol and insulin 24–48 h after calving. The light programmes used were 9.5 L:14.5 D and 17.5 L:6.5 D. During 4–8 weeks before calving one group was on the long day programme (LL group) and the other on the short day programme (DL group) in both experiments. After calving, both groups were on the long day programme. Indoor light intensity was 150 lux at the eye level of the cows in both experiments. Outdoor light intensity during the summer was also included in the calculationsln the second experiment.

In the first study, with 6 cows and 6 first calvers, the plasma concentration of prolactin increased significantly in the LL group but not in the DL group during 30-3 days before calving (b = 1.88±0.70 μg/l/d, p<0.01). During weeks 1–8 after calving there was a similar increase in the DL group (b _plasma = 12.03 μg/l/w, p < 0.001 and b _milk = 0.88±0.37 μg/l/w, p <0.05), but no significant change in the LL group. The first calvers of the LL group produced 10–12% more energy-corrected milk (3.14 MJ/kg) and kg milk fat than those of the DL group (p<0.05). No production traits of the cows were influenced by the light programmes (p>0.40), but both groups of cows were severely affected by illness after calving. Melatonin, growth hormone and Cortisol concentrations in plasma showed no significant daily variation and no effect of light programmes (p>0.45). There was no significant difference in the insulin concentration between groups (p = 0.28), although the group on long days had lower concentrations at all hours. The concentration of plasma prolactin was lower during the dark hours than during the light hours (p<0.05). In the second experiment, 36 first calvers were stalled 6–8 weeks before expected calving following a period of grazing. The stalling involved a substantial decrease in daily maximum light intensity for the cows. From the first day in experiment to 10 days before expected calving, the plasma prolactin concentration decreased from 41.5 ± 3.7 to 29.0 ± 3.3 μg/l, p<0.05, equally in both groups. After calving, the plasma prolactin concentration increased in the DL group (b=4.20 ± 1.93 μg/l/w, p<0.05) to a higher level than at the start of the experiment. There was no difference in milk production (p>0.50).

The results indicate that a continuously increasing secretion of prolactin in the last month of gestation may promote milk production in early lactation.     

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.     

Effect of fractionated weaning on hormonal patterns and weaning to oestrus interval in primiparous sows

The effect of weaning the 4-5 heaviest piglets in the litter on day 33 of lactation and the remainder 2 days later (fractionated weaning) on plasma levels of prolactin, cortisol, oestradiol-17 beta (E2), progesterone (P4) and LH, as well as on the weaning to oestrus interval in primiparous sows was studied. Twelve crossbred sows were grouped into 6 pairs according to farrowing date and litter size. The litter of 1 sow in each pair (F) was weaned in 2 stages, and the other conventionally weaned at 35 days (C). Blood samples were collected via a permanent jugular vein catheter every 3 h from 9 a m to 9 p m daily throughout the experimental period, and intensively at 15 min intervals for 12 h on the day of first and final weaning and for 6 h on the day after each weaning. All sows were slaughtered following their first post-weaning oestrus and the reproductive organs were macroscopically examined. Lactational oestrus was not observed in any of the sows. Sows from 5 out of 6 pairs showed oestrus within 8 days of weaning and post-mortem examination showed normal ovulation. There was a tendency for the F sows to have a shorter weaning to oestrus interval, as compared with the C sows (5 of 6 pairs, 4.8 days v 5.6 days). The plasma levels of prolactin around weaning were not significantly different between the 2 groups. Within 6 h after final weaning, the prolactin concentrations decreased gradually from 7.6 and 8.7 to 1.6 and 1.7 microgram/l in the control and treatment groups, respectively. The plasma levels of cortisol, showing a diurnal rhythm (with the lowest level at 6 and/or 9 p m), did on no occasion differ between the 2 groups. On the day of final weaning, no diurnal rhythm was observed, with cortisol remaining high at 6 and 9 p m. The plasma levels of E2 and P4 were low until final weaning in both groups. After final weaning the E2 levels rose faster in the F sows than in the C sows, to 44.3 and 34.8 pmol/l, respectively, on day 2 (p less than 0.01). No significant differences in levels of plasma LH and the number of LH pulses were observed between the groups. After final weaning the average and base levels of LH and the number of LH pulse(s) increased significantly.     

Sequential Hormonal Changes in the Postpartum Dairy Cow

Peripheral plasma levels of 15-keto-13,14-dihydro-PGF_2α, progesterone, Cortisol, LH and prolactin were studied in 6 primiparous postpartum dairy cows. The cows were followed by hormone measurements and clinical examinations from parturition until pregnancy was established. Blood was collected 3 times per day. The cervix, uterus and ovaries were examined by rectal palpation at 6–10 days intervals. The cows were observed for signs of oestrus twice daily and were additionally teased with a bull to provoke standing heat.Four cows had a normal parturition and dropped their fetal membranes shortly afterwards. (NR group). The remaining 2 retained their fetal membranes for more than 24 h following parturition (RFM group). One out of 6 cows showed standing oestrus at the first ovulation, 4 animals were in oestrus at the second ovulation and all cows showed signs of oestrus at the third ovulation. Although the length of the first luteal phase varied from 9 to 22 days a corpus luteum was in all cases palpated. The secretion of progesterone during the first luteal phase was terminated by a PGF_2α release.A significant difference in 15-keto-13,14-dihydro-PGF_2α levels between the 2 groups was found on days 0–4 (2.39 vs 6.87 nmol/1 at Ρ < 0.06). Postpartum prostaglandin F_2α release as reflected by the level of 15-keto-13,14-dihydro-PGF_2α lasted shorter in the NR group than in the RFM group (15–17 vs 21 days). Significant positive correlations between 15-keto-13,14-dihydro-PGF_2α and Cortisol as well as between prolactin and Cortisol during the first 24 days postpartum were noted only in cows having normal parturition. The most pronounced daily prolactin variations occurred during the second luteal phase (NR group), when a significant difference between the times 8.00, 12.00 and 15.00 was recorded (14.7, 31.5 and 19.7 μg/l, respectively). Moreover, a partial negative correlation between log value of prolactin and arithmetical value of LH was found in these cows only during the first luteal phase after parturition.     

Plasma Cortisol and Behaviour in Early Weaned Piglets

Comparisons were made of Cortisol concentration and behavioural activity between piglets weaned at 3 weeks of age (early weaned piglets), and piglets weaned at 8 weeks of age (control piglets). Early weaned littermates were placed in cages with a floor area of either 0.20 m² per piglet or 0.15 m² per piglet. At 8 weeks of age, the piglets in the control litters were moved from the sow and early weaned piglets were moved from the cages. All litters were placed in pens, 1 pen for each litter. Radioimmunoassay was used for plasma Cortisol determination. Behavioural activity included determination of play, aggressive behaviour and non-nutritive oral activity. Statistical analysis for possible association between plasma Cortisol concentration and age, housing conditions, litter and sequence of capture of littermates for blood collection revealed a complex pattern. Generally, higher plasma Cortisol concentrations and more aggressive and non-nutritive oral activity, but less playing activity, were associated with early weaning. Plasma Cortisol concentration in individual piglets was positively correlated to both aggressive and non-nutritive oral behaviour, but was negatively correlated to playing behaviour. It is concluded that early weaning is a stressor.     

Use of a low-dose ACTH stimulation test for diagnosis of hypoadrenocorticism in dogs

BACKGROUND: Although definitive diagnosis of hypoadrenocorticism usually is made by an adrenocorticotrophic hormone (ACTH) stimulation test using 250 microg/dog of synthetic ACTH (cosyntropin/tetracosactrin), increased costs have prompted a search for less-expensive diagnostic methods. HYPOTHESIS: A low-dose ACTH stimulation test (5 microg/kg) will distinguish between dogs with nonadrenal illness and hypoadrenocorticism. Additionally, administration of cosyntropin will not affect the results of another ACTH stimulation test performed 24 hours later. ANIMALS: Eight healthy adult dogs and 29 hospitalized dogs with suspected hypoadrenocorticism. METHODS: In this prospective study, each healthy dog received 4 ACTH stimulation tests. Dogs received either 5 microg/kg or 250 microg/dog of cosyntropin on day 1 and the alternate dose on day 2. The opposite dosing sequence was used after a 2-week washout period (days 15 and 16). Dogs with suspected Addison's disease received 2 ACTH stimulation tests, 24 hours apart, using either a dose of 5 microg/kg cosyntropin or 250 microg/dog on the 1st day and the alternate dose on the 2nd day. RESULTS: In healthy dogs, poststimulation cortisol concentrations on days 2 and 16 and days 1 and 15 were equivalent (90% confidence interval [CI]: 86.7-101.2%). In dogs with suspected Addison's disease, mean (+/-SD) cortisol responses to ACTH in the 5 microg/kg dose (16.2+/-7.7 microg/dL) and 250 microg/dog dose (15.9+/-6.3 microg/dL) were statistically equivalent (90% CI: 91.2-105.4%). CONCLUSIONS AND CLINICAL IMPORTANCE: Low-dose ACTH stimulation testing distinguishes between dogs with nonadrenal illness and hypoadrenocorticism. Additionally, the administration of 2 ACTH stimulation tests on consecutive days does not affect results of the second test.