Plasma pro-opiomelanocortin, pro-adrenocorticotropin hormone, and pituitary adenoma size in dogs with Cushing's disease

It is difficult to predict the size of pituitary corticotroph tumors in dogs with Cushing's disease (pituitary-dependent hyperadrenocorticism [PDH]) without pituitary imaging techniques. The purpose of this study was to examine the relationship between plasma adrenocorticotropin hormone (ACTH) precursor concentration and pituitary size in dogs with Cushing's disease. Plasma concentrations of ACTH precursors (pro-opiomelanocortin [POMC]/pro-ACTH) and pituitary tumor height/brain area were measured in 36 dogs with pituitary corticotroph adenomas of various sizes. There was a correlation between tumor size (measured as the pituitary tumor height/brain area ratio [P/B]) and POMC/pro-ACTH concentration (r = .70; P < .0001). Dogs with P/B > or = 0.40 x 10(-2) mm(-1) had higher concentrations of ACTH precursors than dogs with P/B < 0.40 x 10(-2) mm(-1) (median concentration 85 pmol/L, range 15-1,350 pmol/L, n = 14 versus 15 pmol/L, range 15-108 pmol/L, n = 22; P < .0001). With a threshold of 35 pmol/L of POMC/pro-ACTH concentration, the estimated sensitivity and specificity of the kit were 93% (95% confidence interval [CI], 79-100%) and 86% (95% CI, 73-100%), respectively. We interpret these data as indicating that measurement of POMC and pro-ACTH might be of value in the characterization of tumor size in dogs with Cushing's disease. Low POMC/pro-ACTH concentrations make it unlikely that a large pituitary tumor exists in dogs with PDH.     

Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. I. Effects on the immune axis when fed diets containing spray-dried plasma

The objective of the present study was to evaluate the potential immunological benefit of adding menhaden fish oil to the diet of weaned pigs. Twenty-four crossbred male pigs were weaned at approximately 18 days of age and placed on a complex nursery diet containing 30% lactose and 7% plasma protein with 6% corn oil as the fat source (Cont, n=12) or with 5% menhaden fish oil and 1% corn oil as the fat source (MFO, n=12) for a period of 15 days. Body weights did not differ (P>0.78) between dietary groups either at the beginning or end of the 15 days feeding period. On day 15, all pigs were non-surgically fitted with an indwelling jugular catheter. On d 16, pigs received an i.v. injection of either saline (n=6/dietary group) or lipopolysaccharide (LPS; 150 μg/kg body weight; n=6/dietary group) and blood samples were collected at 30 min intervals for a period of 5 h. Serum was harvested and stored at −80 °C for analysis of cortisol (CS), corticosteroid-binding globulin (CBG), tumor necrosis factor-alpha (TNF-) and interferon-gamma (IFN-γ). There was no significant effect of diet on basal concentrations (Time 0) of any of the blood parameters analyzed. A Time×Treatment×Diet interaction (P<0.03) was observed for serum CS such that those pigs which consumed the MFO diet followed by LPS treatment had a reduced CS response as compared to the LPS-treated pigs on the Cont diet. A Time×Treatment interaction (P<0.01) was observed for serum CBG such that LPS treatment reduced circulating CBG as compared to the saline-treated pigs. Time×Treatment×Diet interactions were also observed for serum concentrations of TNF- (P=0.084) and IFN-γ (P=0.022) such that both the TNF- and IFN-γ response to the LPS challenge was lower in those pigs receiving the MFO diet as compared to the LPS-treated pigs on the Cont diet. Overall, serum CS was negatively correlated with the CBG response (r=−0.40, P<0.001), however, the strongest negative correlation was observed in the LPS-treated pigs which consumed the MFO diet (r=−0.63, P<0.001). While further studies are needed to evaluate the immunological response of including MFO in the nursery pig diet, the present study demonstrates that supplementation with MFO does indeed alter the immunological response to an LPS challenge.     

Skatole metabolism in the intact pre-pubescent male pig: The relationship between hepatic enzyme activity and skatole concentrations in plasma and fat

The objective of this study was to evaluate, in the pre-pubescent intact male pig, the relationship between skatole levels and the activity of hepatic cytochrome P4502E1 (CYP2E1), cytochrome P4502A (CYP2A), aldehyde oxidase (AO), and phenol sulfotransferase 1A1 (SULT1A1). The activity of these enzymes has been positively associated with skatole clearance in mature boars. Twenty-four intact male pigs were weaned at 28 days of age and slaughtered 2 weeks postweaning, at which time caecal contents, blood, fat, and liver samples were collected. Caecal contents and fat were analyzed for skatole concentrations, and plasma was analyzed for skatole and steroid hormone (testosterone (T), dehydroepiandrosterone (DHEAS), estrone sulphate (E₁S)) concentrations. CYP2A, CYP2E1, and AO, as well as SULT1A1 activities were evaluated in liver samples. Stepwise regression was utilized considering plasma or fat skatole concentration as the dependent variables and hormone concentrations and enzyme activities as independent variables. The activities of the enzymes CYP2A, CYP2E1, and AO and concentrations of the hormones T, DHEAS, or E₁S were not correlated with concentrations of skatole in plasma or fat. However, SULT1A1 activity was negatively correlated with plasma (r = − 0.70, P < 0.05) and backfat (r = − 0.41, P < 0.05) skatole concentrations. Furthermore, this correlation was improved in plasma (r = − 0.88, P < 0.05) and fat (r = − 0.63, P < 0.05) when the concentrations of skatole in caecal contents was included as an independent variable in the multiple regression analysis, demonstrating the importance of measuring skatole production in these studies. T, DHEAS, and E₁S concentrations in plasma were not correlated with the activity of any of the enzymes evaluated. This study suggests that SULT1A1 is important in the metabolism of skatole in pre-pubescent pigs and the overall metabolism of skatole in the pre-pubescent pig differs from that in the mature boar.     

Prolactin signaling in porcine adrenocortical cells: involvement of protein kinases

Prolactin (PRL) was found to have a stimulatory effect on adrenal steroidogenesis in vivo and in vitro in several species including pigs. PRL signal transduction pathways, however, in adrenocortical cells are poorly recognized. Therefore, the goal of this paper is to ascertain the involvement of protein kinase C (PKC) and tyrosine kinases in PRL signaling in porcine adrenal cortex. Adrenals were harvested from locally slaughtered mature gilts. Cortical cells were dispersed by sequential treatment with collagenase. The cells were seeded into 24-well culture plates at a density of 3×10⁵/mL. Cells were incubated with or without PRL (500 ng/mL), ACTH (5 nM—a positive control), tyrosine kinase inhibitor—genistein (1; 2.5 or 5 μM), PKC inhibitor—sphingosine (20–1000 nM) and PKC activators—diacylglycerol (DiC8; 10–100 μM) and phorbol ester (PMA; 1–1000 nM). All incubations were performed for 8 h (95% air and 5% CO₂, 37°C). PRL and ACTH (P<0.05) increased cortisol and androstenedione (A₄) secretion. DiC8 and PMA mimicked the stimulatory effect of PRL. Sphingosine (P<0.05) suppressed basal and PRL-stimulated steroid secretion. Genistein inhibited (P<0.05) PRL-stimulated cortisol secretion and enhanced (P<0.05) basal and PRL-stimulated A₄ secretion. Moreover, PKC activation was assessed by measuring the specific association of [³H]phorbol dibutyrate ([³H]PDBu) with adrenocortical cells after treatment with PRL or ionomycin (a positive control). PRL (within 2–3 min) and ionomycin (within 2–5 min) increased (P<0.05) specific binding of [³H]PDBu to the porcine adrenocortical cells. In addition, PRL did not augment the cortisol and A₄ secretion by PKC-deficient adrenocortical cells. In conclusion, presented results support the hypothesis that PKC and tyrosine kinases are involved in PRL signaling in adrenocortical cells in pigs. Moreover, activation of PKC is associated with the increased secretion of cortisol and A₄.     

Growth performance and metabolic responses in barrows fed low-protein diets supplemented with essential amino acids

This study was conducted to determine the effect of supplementation with Lys, Met, Thr to low-protein diets on growth performance and metabolic responses in growing barrows. Seventy crossbred barrows (Duroc × Yorkshine), with an average initial body weight of 16 kg, were fed diets containing five crude protein (CP) levels (18.2, 16.5, 15.5, 14.5, and 13.6%) and 0.83% true ileal digestible lysine. On d 17 of the trial, all pigs were fitted with an indwelling jugular catheter for blood collection. On d 18 and 53, blood samples were obtained at 4 h post feeding. Reducing dietary CP concentration linearly decreased (P < 0.05) ADG, ADFI, feed efficiency, longissimus muscle area (LMA), and plasma concentrations of urea N and protein, while increasing backfat thickness linearly (P < 0.05). Pigs fed the 13.6% CP diet exhibited the poorest growth performance. On d 53, dietary CP affected plasma IGF-I (quadratic, P < 0.01) and leptin (linear, P < 0.01) concentrations. Plasma leptin levels were correlated positively with backfat thickness (r = 0.76, P < 0.05) and negatively with LMA (r = − 0.64, P < 0.01). There was a positive correlation between ADG and plasma IGF-I concentrations (r = 0.91, P < 0.05). Our results suggest that adequate provision of nonessential AA and all EAA is required for achieving maximum growth performance and reducing fat accretion in growing pigs fed low-CP diets.     

Correlation between carcass conformation and fat cover degree, and muscle fatty acid profile of yearling bulls depending on breed and mh-genotype

The objective was to study the relationships between the actual European beef carcass classification scale, which classifies carcasses with regard to conformation and degree of fat cover scores, and muscle fat quality, depending on breed and mh-genotype. For this purpose samples from 100 yearling bulls from “Asturiana de los Valles” (24 AV(mh/mh), 26 AV(mh/+), 25 AV (+/+)) and “Asturiana de la Montaña” (25 AM) were analysed. The results of the study showed that breed or genotype affect carcass measurement scores and muscle fatty acid profile through its important effect on animal overall fatness. Homozygous double-muscled animals produced carcasses with high conformation and low intramuscular (IM) fat content. While early-maturing and rustic AM animals produced low carcass yield and high IM fat content. The other genotypes (mh/+, +/+) showed, in general, intermediate characteristics. Referring to correlations, carcass conformation was negatively related to saturated (SFA) (r = − 0.69, P < 0.001) and monounsaturated fatty acid (MUFA) (r = − 0.69, P < 0.001) groups, and positively to polyunsaturated (PUFA) (r = 0.72), n-6 (r = 0.72), n-3 (r = 0.71) and unsaturated fatty acid (UFA) (r = 0.69) groups, being all of them significant (P < 0.001). However, carcass degree of fat cover was positively related to SFA (r = 0.53, P < 0.001) and MUFA (r = 0.62, P < 0.001), and negatively to PUFA (r = − 0.61), n-6 (r = − 0.60), n-3 (r = − 0.62) and UFA (r = − 0.53) groups, being all of them significant. Moreover, simple and low-cost prediction equations were calculated for a rapid and sufficiently accurate fatty acid group (SFA, MUFA, PUFA, n-6, n-3, UFA) estimation (R² > 0.46, P < 0.001). In general, meat obtained from double-muscled animals display a more appropriate IM fatty acid profile from the nutritional point of view according to actual recommendations, but it could happen the disability of these lean animals to deposit sufficient IM fat to ensure consumer overall liking or acceptability.     

Administration of recombinant porcine somatotropin (rpST) changes hormone and metabolic status during early pregnancy

The objective of this study was to examine the effects of somatotropin (ST) on porcine reproductive and metabolic statuses during early pregnancy. Four pregnant crossbred gilts received 6 mg of recombinant porcine somatotropin (rpST) daily from days 10 to 27 after artificial insemination while six pregnant gilts served as controls. Blood samples were taken on days 8, 10, 12, 14, 18, 22, and 27 prior to rpST injections (8:00 h) and subsequently at 9:00, 10:00, 12:00, 14:00, 16:00, 18:00, and 20:00 h. On all remaining days of treatment, samples were taken once daily before injections (8:00 h). The samples were assayed for the metabolic hormones: ST, insulin-like growth factor I (IGF-I), insulin, thyroxine (T₄), triiodothyronine (T₃), and cortisol; for metabolites: free fatty acids (FFA) and glucose; and for the reproductive hormones: luteinizing hormone (LH), progesterone, estradiol-17β, estrone sulfate, and prostaglandin F₂. Delivery of rpST daily induced a 20- to 40-fold increase in plasma ST concentrations. Moreover, repeated administration of rpST resulted in a continuous increase in plasma IGF-I concentration (P<0.001), from 191.0±22.3–340.0±15.3 ng/mL 24 h after initial injection to 591.3±46.8 ng/mL after final injections. Mean serum insulin tended to be greater in rpST-treated gilts. Blood concentrations of T₄ were reduced (P<0.05) from day 14 of gestation in treated gilts while T₃ concentrations remained unchanged. Concentrations of both glucose and FFA were greater (P<0.01) and cortisol concentrations were unchanged in treated gilts. Changes in reproductive steroid hormones were minimally affected. Circulating progesterone (P=0.078), and estradiol-17β (P=0.087) concentrations tended to be lower in treated animals. These data show that treatment of pregnant gilts with rpST during early gestation mainly impacts metabolic rather than reproductive status.     

Effect of progressive cachectic parasitism and growth hormone treatment on hepatic 5'-deiodinase activity in calves

Thyroid status is compromised in a variety of acute and chronic infections. Conversion of thyroxine (T₄) into the metabolically active hormone, triiodothyronine (T₃), is catalyzed by 5′-deiodinase (5′D) mainly in extrathyroidal tissues. The objective of this study was to examine the effect of protozoan parasitic infection (Sarcocystis cruzi) on hepatic 5′D (type I) activity and plasma concentrations of T₃ and T₄ in placebo- or bovine GH (bGH)-injected calves. Holstein bull calves (127.5±2.0 kg BW) were assigned to control (C, ad libitum fed), infected (I, 250,000 S. cruzi sporocysts per os, ad libitum fed), and pair-fed (PF, non-infected, fed to intake of I treatment) groups placebo-injected, and three similar groups injected daily with pituitary-derived bGH (USDA-B-1, 0.1 mg/kg, i.m.) designated as C_GH, I_GH and PF_GH. GH injections were initiated on day 20 post-infection (PI), 3–4 days prior to the onset of clinical signs of the acute phase response (APR), and were continued to day 56 PI at which time calves were euthanized for liver collection. Blood samples were collected on day 0, 28, and 55 PI. Alterations in nutritional intake did not affect type I 5′D in liver. Treatment with bGH increased (P<0.05) 5′D activity in C (24.6%) and PF (25.5%) but not in I calves. Compared to PF calves, infection with S. cruzi reduced 5′D activity 25% (P<0.05) and 47.8% (P<0.01) in placebo- and bGH-injected calves, respectively. Neither nutrition nor bGH treatment significantly affected plasma concentrations of T₄ and T₃ on day 28 and 55 PI. However, plasma thyroid hormones were reduced by infection. On day 28 PI, the average plasma concentrations of T₃ and T₄ were reduced in infected calves (I and I_GH) 36.4% (P<0.01) and 29.4% (P<0.05), respectively, compared to pair-fed calves (PF and PF_GH). On day 55 PI, plasma T₃ still remained lower (23.7%, P<0.01 versus PF) in infected calves while plasma T₄ returned to control values. The data suggest that parasitic infection in growing calves inhibits both thyroidal secretion and extrathyroidal T₄ to T₃ conversion during the APR. After recovery from the APR, thyroidal secretion returns to normal but basal and bGH-stimulated generation of T₃ in liver remains impaired.     

Endotoxin challenge increases xanthine oxidase activity in cattle: effect of growth hormone and vitamin E treatment

In addition to its basic role in the metabolism of purine nucleotides, xanthine oxidoreductase (XOR) is involved in the generation of oxygen-derived free radicals and production and metabolic fate of nitric oxide (NO). Growth hormone (GH) and Vitamin E (E) have been shown previously to modify immune response to infection. Our objective was to determine in heifers the effect of endotoxin challenge (LPS; 3.0 μg/kg BW, i.v. bolus, Escherichia coli 055:B5) on xanthine oxidase (XO) activity in plasma and liver and the modification of this response by daily treatment with recombinant GH (0.1 mg/kg BW, i.m., for 12 days) or GH+E (E: mixed tocopherol, 1000 IU/heifer, i.m., for 5 days). In experiment 1, 16 heifers (348.7±6.1 kg) were assigned to control (C, daily placebo injections), GH, or GH+E treatments and were challenged with two consecutive LPS injections (LPS1 and LPS2, 48 h apart). After LPS1, plasma XO activity increased 290% (P<0.001) at 3 h, reached peak (430%) at 24 h and returned to basal level by 48 h after LPS2. XO responses (area under the time×activity curve, AUC) were greater after LPS1 than LPS2 (P<0.001). Total plasma XO responses to LPS (AUC, LPS1+LPS2) were augmented 55% (P<0.05) over C with GH treatment but diminished to C responses in GH+E. There was a linear relationship (r²=0.605, P<0.001) between total response in plasma XO activity and plasma nitrate+nitrite concentration. In experiment 2, 24 heifers (346±6 kg) were assigned to C or GH treatments and liver biopsy samples were obtained at 0, 3, 6, and 24 h after a single LPS challenge. Hepatic XO activities increased 63.3% (P<0.05) 6 h after single LPS challenge and remained elevated at 24 h (100.1%, P<0.01) but were not affected by GH treatment. Results indicate that LPS-induced increases in plasma XO activity could be amplified by previous GH treatment but attenuated by E administration. The data also suggest that E may be effective in controlling some mediators of immune response associated with increased production of NO via the effect on XO activity and its production of superoxide anion as well as uric acid.     

Influence of gonadotropins on insulin- and insulin-like growth factor-I (IGF-I)-induced steroid production by bovine granulosa cells

To determine the effect of gonadotropins on insulin- and insulin-like growth factor (IGF-I)-induced bovine granulosa cell functions, granulosa cells from bovine ovarian follicles were cultured for 2 days in the presence of 10% fetal calf serum (FCS), and then cultured for an additional 2 days in serum-free medium with added hormones. In the presence of 0 or 1 ng/mL of insulin or IGF-I, FSH had little or no effect (P>0.05) on estradiol production by granulosa cells from both small (1–5 mm) and large (≥8 mm) follicles. However, in the presence of ≥3 ng/mL of insulin, FSH increased (P<0.05) estradiol production by granulosa cells from small and large follicles such that the estimated dose (ED₅₀) of insulin necessary to stimulate 50% of the maximum estradiol production was decreased by 2- to 3-fold from 22 to 28 ng/mL in the absence of FSH to 7–14 ng/mL in the presence of FSH. Similarly, in the presence of ≥3 ng/mL of IGF-I, FSH increased (P<0.05) estradiol production by granulosa cells from small and large follicles such that the ED₅₀ of IGF-I for estradiol production was decreased by 4- to 5-fold from 25 to 36 ng/mL in the absence of FSH to 5–6 ng/mL in the presence of FSH. In the presence of FSH, the maximal effect of insulin on estradiol production was much greater than that of IGF-I (137- versus 12-fold increase) and were not additive; when combined, 100 ng/mL of IGF-I completely blocked the stimulatory effect of 100 ng/mL of insulin. In the absence of FSH, the maximal effect of insulin and IGF-I on estradiol production was similar. Concomitant treatment with 30 ng/mL of LH reduced (P<0.05) insulin-stimulated estradiol production by 52% on day 1 and 19% on day 2 of treatment. Insulin, IGF-I and FSH also increased (P<0.05) granulosa cell numbers and progesterone production but their maximal effects were less (i.e., <4-fold increase) than their effects on estradiol production. In conclusion, insulin and IGF-I synergize with FSH to directly regulate ovarian follicular function in cattle, particularly granulosa cell aromatase activity.     

Postnatal changes in electromyographic signals during piglet growth, and in relation to muscle fibre types

This study uses non-invasive evoked surface electromyography (SEMG) to investigate postnatal muscle development in pigs, and to assess any correlation between recorded signal parameters and muscle fibre types in two different skeletal muscles. Four litters (n = 43) of Large White × Landrace pigs were used. Evoked SEMG measurements were taken on days 2, 5, 14, 26, 60 and 151 post partum from m. Longissimus dorsi (LD) and on days 14, 26, 60 and 151 post partum from m. Biceps femoris (BF). A third of each litter was slaughtered at days 27, 61 and 153 post partum. Biopsy samples for LD and BF were taken to categorize muscle fibre types. For LD there was a significant increase in compound muscle action potential (CMAP) parameters from day 2 to day 5 post partum, whilst for BF significant increases occurred from days 14 to 26 post partum. Fibre type development in both muscles showed a significant decrease in type IIA fibre number (P < 0.001) and an increase in type IIB fibre number (P < 0.001). Analysis of CMAP parameters in relation to fibre type percentages showed significant positive correlations between percentage of type I fibres in BF and CMAP Area (r = 0.71; P = 0.05), SlopeL (r = 0.79; P = 0.02) and Corr Peak (r = 0.78; P = 0.02) and a negative correlation with SlopeT (r = 0.89; P = 0.003) at day 151. A greater CMAP Area and Corr Peak as well as steeper leading and trailing slopes were on the other hand correlated to fewer type IIB fibres.

It is concluded that 1) changes in the evoked CMAP are muscle-specific during early postnatal development in the pig, and 2) in spite of a small sample size, the correlations between CMAP signal parameters and fibre type percentages warrant further investigation.     

Mammary uptake and excretion of prostanoids in relation to mammary blood flow and milk yield during pregnancy-lactation and somatotropin treatment in dairy goats

Mammary arterious − venous differences (A − V) and excretion into milk of four prostanoids were related to changes in milk yield and milk vein blood velocity (MBV) in goats at different stages of pregnancy and lactation, and during somatotropin (ST) treatment in mid-lactation. Arterial concentrations and mammary A − V for the vasodilators prostacyclin (PGI₂) and prostaglandin (PG) E₂ (measured as 6-keto-PGF₁ and bicyclic PGE₂, respectively) decreased from late pregnancy to lactation. A − V were negatively correlated to MBV (r = −0.32 to −0.34). Arterial concentrations of the vasoconstrictors PGF₂ and TXA₂ (measured as TXB₂) changed similarly, but no A − V across the mammary gland were found. The vasodilator to vasoconstrictor ratio in plasma was around 1:1, and in skimmed milk around 0.29–0.49 due to significantly higher TXB₂ levels in milk compared to plasma. Close linear correlations were established between milk yield and excretion of TXB₂ into milk (r = 0.80, P < 0.001), and between MBV and PGE₂ excretion into milk (r = 0.69, P < 0.001). ST treatment stimulated MBV and mammary prostanoid supply, and decreased prostanoid concentration in milk vein plasma. The high arterial levels of prostaglandins during pregnancy most likely reflected uterine synthesis. Our results support a role for PGI₂ and PGE₂ in local mammary blood flow regulation during lactation. Increased mammary uptake of these two prostanoids may be involved in the mammary blood flow response to ST. TXA₂ may be synthesized by mammary epithelial as well as vascular cells, and TXA₂ may be an important factor in regulation of mammary function.     

Erhualian and Pietrain pigs exhibit distinct behavioral, endocrine and biochemical responses during transport

This study describes the breed-specific coping characteristics of pigs in response to transport stress. The dynamic changes of behavior, the activities of creatine kinase (CK) and lactate dehydrogenase (LDH), as well as the plasma concentrations of stress and metabolic hormones in Erhualian (EHL) and Pietrain (PIE) pigs during 2 h transport were investigated. The majority of the behavior of EHL pigs consisted of oral/nasal/facial (ONF) and fit behaviors during the initial observations (first 15 min after start), and these behaviors were replaced by increased time sitting and lying in later observations (middle and last 15 min of transport). In contrast, PIE pigs showed high levels of ONF behaviors in initial observation, followed by high frequency and duration of standing during the middle and the last observation period. PIE pigs demonstrated significantly higher plasma CK (P < 0.01) and LDH activities (P < 0.05). There were significant effects of time and time × breed interaction (P < 0.05) on CK activities (P < 0.01) in both breeds. Plasma ACTH levels did not differ between breeds, yet a significant effect of time (P < 0.05) was shown during transport. EHL pigs exhibited consistently higher basal and stimulated plasma cortisol levels (P < 0.05). There were significant time effects on metabolic hormones (insulin, T3 and T4) (P < 0.01), whereas no significant breed effect for these hormones were found. These results indicate that different coping strategies apply in EHL pigs, as reflected by different behavioral, endocrine and biochemical responses during transport as compared with PIE pigs.     

The effect of dietary crude protein concentration and inulin supplementation on nitrogen excretion and intestinal microflora from finisher pigs

A 2 × 2 factorial experiment was used to investigate the interaction between dietary crude protein (CP) concentration (200 vs 140 g/kg) and inulin supplementation (0 vs 12.5 g/kg) on nitrogen (N) excretion and intestinal microflora from 16 boars (n = 4, 74.0 kg live weight). The diets were formulated to contain similar concentrations of digestible energy and lysine. Pigs offered the high CP diets had a higher excretion of urinary N (P < 0.01), faecal N (P < 0.01) and total N (P < 0.001) than the pigs offered the low CP diets. Inulin supplementation increased faecal N excretion (P < 0.05) and decreased urine: faeces N ratio (P < 0.05) compared to the inulin free diets. There was no significant effect (P > 0.05) of dietary treatment on N retention. There was an interaction (P < 0.05) between dietary CP concentration and inulin supplementation on caecal E.coli. Pigs offered the diet containing 200 g/kg CP plus inulin decreased the population of E.coli compared to the inulin supplemented 140 g/kg protein diet. However, CP concentration had no significant effect on the population of E.coli in the unsupplemented diets. Inulin supplementation increased caecal bifidobacteria (P < 0.01) compared to the inulin free diets. In conclusion, inulin supplementation favourably altered N excretion and lowered the population of E.coli at high CP concentrations.     

Canine echinococcosis in northwest Libya: assessment of coproantigen ELISA, and a survey of infection with analysis of risk-factors

In order to determine the prevalence and risk factors for canine echinococcosis in different endemic localities in the Tripoli area of northwest Libya, stray dogs were examined post-mortem, and owned dogs screened for Echinococcus granulosus infection using a standardised genus specific coproantigen ELISA. The prevalence of E. granulosus infection at necropsy in stray-dogs was 25.8% (15/58, 95% CI 15.3–39.0%), and 21.6% (72/334, 95% CI 17.3–26.4%) of owned dogs tested were positive by coproantigen ELISA. Sheepdogs appeared to have a significantly higher copro-positive prevalence (19/19 positive, p = 0.003), compared to 23.6% of other dog classes (e.g. 52/220 guard dogs and household pets). Worm burdens in necropsied dogs ranged from 29 to 2900 (mean 1064) and were positively correlated to coproantigen ELISA OD values (r_s = 0.87, p < 0.001), but negatively correlated with dog age (r_s = −0.69, p = 0.001). Dog age was a significant factor in copro-prevalence as there was an increasing coproantigen-positive tendency in younger dogs (≤5 years, p = 0.04). A total of 45/132 (34%, 95% CI 25.9–42.1%) of farms/homestead had at least one dog that was coproantigen positive. Overall copro-prevalence in dogs by locality varied, with Alkhums (Leptis-Magna) district having the highest copro-prevalence at 38.7% (24/62, 95% CI 26.6–50.8%) (p = 0.001). Coproantigen testing of a cohort of owned dogs before and approximately 15 months after praziquantel treatment showed a significant decrease in the coproantigen positive rate from 21.6% (72/334) to 9% (21/233) post-treatment. The overall E. granulosus coproantigen positive rate (‘re-infection rate’) within the same cohort of dogs was 22 % (10/45) by 15 months post-treatment. Significant risk factors for a copro-positive owned dog were associated with non-restraint of dogs, and owners that did not de-worm their dogs. Home slaughtering of livestock and lack of knowledge about E. granulosus transmission were also significant risk factors for a canine coproantigen positive result.     

Seasonal and pulsatile dynamics of thyrotropin and leptin in mares maintained under a constant energy balance

The objective of this study was to determine if seasonal and/or pulsatile variations occur in plasma concentrations of thyrotropin (TSH) and leptin in mares while maintaining a constant energy balance. Blood samples were collected every 20 min during a 24 h period in winter and again in summer from six Quarter Horse type mares. Plasma concentrations of TSH, leptin, and T₄ were determined by radioimmunoassay. No differences were observed in body weight between winter (388.1 ± 12.5 kg) and summer (406.2 ± 12.5 kg; P = 0.11). Plasma concentrations of TSH were greater in the summer (2.80 ± 0.07 ng/ml) when compared to winter (0.97 ± 0.07 ng/ml; P < 0.001). Pulse frequency of TSH was not different between winter (6.17 ± 0.78 pulses/24 h) and summer (5.33 ± 0.78 pulses/24 h; P = 0.49). Mean TSH pulse amplitude, pulse area, and area under the curve were all greater in summer compared to winter (3.11 ± 0.10 ng/ml versus 1.20 ± 0.10 ng/ml, 24.86 ± 0.10 ng/ml min versus 13.46 ± 1.90 ng/ml min, 3936 ± 72.93 ng/ml versus 1284 ± 72.93 ng/ml, respectively; P < 0.01). Mean concentrations of leptin were greater in summer (2.48 ± 0.17 ng/ml) compared to winter (0.65 ± 0.17 ng/ml; P < 0.001). Pulsatile secretion patterns of leptin were not observed in any horses during experimentation. Mean concentrations of T₄ were greater in winter (20.3 ± 0.4 ng/ml) compared to summer (18.2 ± 0.4 ng/ml; P < 0.001). These seasonal differences between winter and summer provide evidence of possible seasonal regulation of TSH and leptin.     

Temporal and spatial patterns of African swine fever in Sardinia

Temporal patterns and spatial distribution of African swine fever (ASF) were studied through the analysis of routinely collected data in the ASF-endemic area of the Province of Nuoro, Sardinia. During 1993–1996, ASF outbreaks were reported from 45 out of the 82 municipalities of the study area. Overall farm-level incidence rate (IR) was 1.3 outbreaks per 100 farms-year. ASF peaked in 1995 (IR=1.8) and declined in 1996 (IR=0.82). Significant (P<0.05) spring peaks of ASF outbreaks and affected municipalities were detected using statistical methods for circular distributions. Spatial clustering of ASF-affected municipalities, as evaluated by join-count statistics, was significant in 1993 (Z_jc=−3.0, P<0.01) and 1994 (Z_jc=−3.2, P<0.01) but not in 1995 (Z_jc=−0.6, P=0.55) and 1996 (Z_jc=−1.2, P=0.23). Extensive pig farming and ASF were spatially co-distributed (κ=0.51, 95% CI=0.33–0.70).     

Effect of constant-release melatonin implants and prolonged exposure to a long day photoperiod on prolactin secretion and hair growth in mouflon (Ovis gmelini musimon)

The aims of this study were to examine whether mouflons exposed to constant long and short day photoperiods are able to exhibit an annual cycle of hair growth and moult, and prolactin (PRL) secretion. Mouflon ewes were assigned to three groups of treatment. Ewes were maintained, either under natural photoperiod (control, n=9), or received a series of subcutaneous melatonin implants from December to April (n=8), or were exposed to a constant long day photoperiod (16-h light:8-h dark; 16L:8D) during 18 months (n=7). Blood was collected weekly to determine PRL concentrations, and hair samples were clipped weekly from the base of the neck to measure the length of predominant hair. Under constant long days and with melatonin implants, mouflons expressed an annual rhythm of PRL secretion, even though these treatments modified the times of rise or falling of PRL concentrations throughout the year. Hair growth initiation was almost coincident with the summer solstice in both control and melatonin-implanted mouflons but occurred two months earlier in long day hold mouflons (P<0.001). Long day hold mouflons had a lower hair growth rate than control and melatonin-implanted mouflons (P<0.001), and at the end of the experiment, a shorter hair length (3.4±0.24 cm; P<0.01) than control (4.3±0.17 cm), and melatonin-implanted mouflons (4.2±0.12 cm). Our data support the conclusion that in mouflon, an endogenous circannual rhythm of PRL secretion exists, and that the seasonal cycle of hair growth and moult appears to depend, at least in part, on circulating levels of PRL.     

I plasma concentrations of adrenocorticotrophic hormone and alpha-melanocyte-stimulating hormone in ferrets (Mustela putorius furo) with hyperadrenocorticism

OBJECTIVE: To determine plasma concentrations of adrenocorticotrophic hormone (ACTH) and alpha-melanocyte stimulating-hormone (alpha-MSH) in healthy ferrets and ferrets with hyperadrenocorticism. ANIMALS: 16 healthy, neutered, privately owned ferrets, 28 healthy laboratory ferrets (21 sexually intact and 7 neutered), and 28 ferrets with hyperadrenocorticism. PROCEDURES: Healthy ferrets were used for determination of reference plasma concentrations of ACTH and a-MSH. Diagnosis of hyperadrenocorticism was made on the basis of history, clinical signs, urinary corticoid-to-creatinine ratios, ultrasonography of the adrenal glands, and macroscopic or microscopic evaluation of the adrenal glands. Blood samples were collected during isoflurane anesthesia. Plasma concentrations of ACTH and alpha-MSH were measured by radioimmunoassay. RESULTS: Plasma concentrations of ACTH in 23 healthy neutered ferrets during the breeding season ranged from 4 to 145 ng/L (median, 50 ng/L). Plasma concentrations of alpha-MSH in 44 healthy neutered or sexually intact ferrets during the breeding season ranged from < 5 to 617 ng/L (median, 37 ng/L). Reference values (the central 95% of the values) for ACTH and alpha-MSH were 13 to 100 ng/L and 8 to 180 ng/L, respectively. Plasma concentrations of ACTH and alpha-MSH in ferrets with hyperadrenocorticism ranged from 1 to 265 ng/L (median, 45 ng/L) and 10 to 148 ng/L (median, 46 ng/L), respectively. These values were not significantly different from those of healthy ferrets. Plasma ACTH concentrations of sexually intact female ferrets in estrus were significantly higher than those of neutered females. CONCLUSIONS AND CLINICAL RELEVANCE: Ferrets with hyperadrenocorticism did not have detectable abnormalities in plasma concentrations of ACTH or alpha-MSH. The findings suggest that hyperadrenocorticism in ferrets is an ACTH and alpha-MSH-independent condition.     

Intrauterine infusion of BQ-610, an endothelin type A receptor antagonist, delays luteolysis in dairy heifers

Three separate in vivo experiments were conducted to evaluate the putative role of endothelin-1 (ET-1) during luteal regression in heifers. In Experiment 1, a single intraluteal injection of 500 μg BQ-610 [(N,N-hexamethylene) carbamoyl-Leu-d-Trp (CHO)-d-Trp], a highly specific endothelin A (ET_A) receptor antagonist, did not diminish the decline in plasma progesterone following a single exogenous injection of 25 mg prostaglandin F2 alpha (PGF₂) administered at midcycle of the estrous cycle. In Experiment 2, six intrauterine infusions of 500 μg BQ-610 given every 12 h on days 16–18 delayed spontaneous luteolysis, as evidenced by an extended elevation (P = 0.054) of plasma progesterone concentration. In Experiment 3, heifers were administered six intrauterine infusions of BQ-610 or saline on days 16–19, and peripheral blood samples were collected from day 11 to 16 (before infusion), hourly on days 16–19 (during infusion), and on days 20–25 (after infusion). BQ-610 treated heifers had markedly higher (P < 0.0001) levels of plasma progesterone compared with saline controls, and this effect was most notable during the infusion period (treatment by period interaction; P ≤ 0.05). Heifers infused with BQ-610 also had higher progesterone levels on day 21 (treatment by time interaction; P ≤ 0.05). Mean plasma concentrations of 13,14-dihydro-15-keto-PGF₂ (PGFM), the primary metabolite of PGF₂, were measured in the samples collected hourly and were not different (P ≥ 0.05) between treatments. These results indicate that the in vivo antagonism of the ET_A receptor can delay functional luteolysis, and supports the theory that ET-1 regulates luteal function in ruminants.