Melatonin and CIDR improved the follicular and luteal haemodynamics,uterine and ovarian arteries vascular perfusion,ovarian hormones and nitric oxide in cyclic cows

This study hypothesizes that melatonin with exogenous progesterone (CIDR) can improve follicular, luteal, ovarian and uterine haemodynamic of heat-stressed cows. Holstein cows (N = 12) studied for two spontaneous oestrous cycles during winter then divided equally during summer into the CIDR group received CIDR for 7 days and the melatonin group (Mel) received three injections of melatonin (75 mg/head) at the CIDR insertion, removal and ovulation days. Blood samples were collected to assay oestradiol (E2), progesterone (P4) and nitric oxide (NO). On day 0 (Ovulation), Mel had more small follicles (p < .05), higher ipsilateral and contralateral ovarian arteries (Ov.A.) peak systolic velocity (PSV), higher ipsilateral uterine artery (Ut.A.) PSV (p = .031) and blood flow volume (BFV), also Mel elevated contralateral Ut.A. PSV and BFV (p < .0001) but lowered contra Ut.A. pulsatility index (PI, p < .0001), E2 (p < .01) and NO (p < .0001). Mel increased the corpus luteum diameter (CL, p < .001), coloured area (p < .007) and P4 (p < .0001) on day 5 and reduced them (p < .05; p < .01) on Day 14. On day 10, Mel obtained CL diameter (p < .03) and coloured area (p < .002) of spontaneous that was higher than CIDR and decreased P4 (p < .003). Mel increased CL diameter, area and coloured area and decreased them thereafter. Mel increased the ipsilateral ovarian and uterine arteries PSV and BFV before ovulation and until day 8. Mel increased P4 and decreased NO until days 6 and 14. In conclusion, the improvement in follicular, luteal, ovarian and uterine haemodynamic and the decrease of NO production proved our hypothesis Melatonin doses higher than 75 mg/head is recommended to improve the heat-stressed cow's fertility.     

IGF-I,GHR and UCP mRNA expression in the liver and muscle of high- and low-feed-efficiency laying Japanese quail at different environmental temperatures

In this study, we analyzed insulin-like growth factor I (IGF-I), growth hormone receptor (GHR) and uncoupling protein (UCP) mRNA expression in the muscle and liver of high- (0.23 g/g) and low- (0.17 g/g) feed-efficiency (FE) Japanese quail at three different air temperatures: comfortable (25 °C), heat stress (38 °C) for 12 h or cold stress (10 °C) for 12 h. Total RNA was extracted from the liver and breast muscle of each quail, and cDNA was amplified using specific primers for the target genes. Expression was analyzed using quantitative real-time PCR (qRT-PCR). IGF-I mRNA expression was higher in the livers of high-FE quail than in the livers of low-FE quail under both heat and cold stress conditions. High-FE birds also showed higher GHR mRNA expression independent of temperature. UCP mRNA expression in the liver was lower in high-FE birds and higher under heat stress compared with the other conditions. IGF-I mRNA expression was higher in the muscle of high-FE quail under the three conditions tested, and UCP mRNA expression was higher under cold stress. Our results suggest that air temperature affects the expression of genes related to growth and mitochondrial energy production, and quail with different feed efficiencies respond differently to environmental stimuli.     

Nocturnal melatonin concentrations vary dramatically between the two jugular veins in most individual sheep maintained under mimicked or natural photoperiod

Two experiments were designed to determine if melatonin concentrations differ between jugular veins. In a first experiment, blood was collected continuously every 30 min from each jugular during 12-h from 6 ewes. In a second experiment, 100 ewes were sampled twice at night simultaneously from the two jugular veins. In both experiments, mean melatonin concentrations were similar between right and left jugulars. However, within individuals, large differences were observed between the two sides (p < 0.001). This difference was stable over time and the higher side varies among individuals. The concentrations of prolactin and oxytocin, measured in the same samples did not show such differences. This observation raises the question of the origin of this phenomenon. Moreover, it has important implications for studies in which melatonin concentrations need to be assessed accurately. Indeed, a reliable quantitative assessment of melatonin production by the pineal gland requires sampling from both jugular veins.     

A comparison of the effects of hydromorphone HCl and a novel extended release hydromorphone on arterial blood gas values in conscious healthy dogs

The purpose of this study was to evaluate arterial blood gases in dogs that were given hydromorphone or extended release liposome-encapsulated hydromorphone (LEH). Dogs were randomly administered LEH, n = 6, (2.0 mg kg⁻¹), hydromorphone, n = 6, (0.2 mg kg⁻¹) or a placebo of blank liposomes, n = 3, subcutaneously on separate occasions. Arterial blood samples were drawn at serial time points over a 6-h time period for blood gas analysis. There was no change from baseline values in P_aCO₂, P_aO₂, (HCO₃-), pH, and SBEc in the dogs that received the placebo. Administration of hydromorphone resulted in significant increases in P_aCO₂ (maximum (mean + SD] 44.4 + 1.1 mm of Hg) and significant decreases in P_aO₂ (minimum (mean + SD) 82.4 + 4.7 mm of Hg) and pH (minimum (mean + SD) 7.31 + 0.01) compared with baseline. Administration of LEH resulted in significant increases in P_aCO₂ (maximum (mean + SD) 44.6 + 0.9 mm of Hg) and significant decreases in P_aO₂ (minimum (mean + SD) 84.8 + 2.6 mm of Hg) and pH (minimum (mean + SD) 7.34 + 0.02) compared with baseline. There was no significant difference between these two groups at any time point. The changes observed in P_aCO₂, P_aO₂, and pH, however, were within clinically acceptable limits for healthy dogs. LEH was determined to cause moderate changes in arterial blood gas values similar to those caused by hydromorphone.     

Banding or Burdizzo castration and carprofen administration on peripheral leukocyte inflammatory cytokine transcripts

The objective was to investigate if Banding or Burdizzo castration of bulls would alter the gene expression profile of a range of peripheral leukocyte inflammatory cytokines (IL-1, IL-6, IL-8, IL-10, interferon-γ and tumor necrosis factor-α) and to determine if the administration of carprofen (C) before castration would affect the expression of these genes. Thirty Holstein-Friesian bulls (5.5 months; Mean 191 ± (SEM) 3.7 kg) were blocked by weight and randomly assigned to one of five treatments: (1) untreated control (CON); (2) Banding castration at 0 min (BAND); (3) BAND following an i.v. injection of 1.4 mg/kg BW of carprofen (C) at −20 min (BAND + C); (4) Burdizzo castration at 0 min (BURD); or (5) BURD following 1.4 mg/kg BW of carprofen at −20 min (BURD + C). Blood samples were collected at 1 h before castration and 6, 24 and 48 h post-castration for routine hematology and quantitative real-time PCR analysis of cytokine gene expression analysis. Generally, there were no differences (P > 0.05) among treatment groups in hematological variables following castration. Cortisol concentrations were unchanged throughout the experimental period in CON bulls. BURD animals had greater cortisol concentrations than BAND and CON animals at 6 h post treatment. Transitory effects were observed only in the expression of IL-6 and TNF-α. The relative expression of IL-6 was greater in the BURD than in the BAND treatment (P < 0.05) at 24 h post-castration and was greater in the BURD + C group than in the BURD group (P < 0.05) at 48 h. The relative expression of TNF-α was greater in BAND than in the BURD group (P < 0.05) at 48 h. In conclusion, these findings indicate that Banding or Burdizzo castration did not have any major effect on peripheral leukocyte inflammatory cytokine gene expression; Banding castration caused a greater pro-inflammatory cytokine gene expression reaction than Burdizzo castration and carprofen administration can affect IL-6 gene expression levels in BURD castrated animals.     

Regulating effects and mechanisms of Chinese medicine decoction on growth and gut hormone expression in heat stressed pigs

Exposure to high temperatures during the summer renders pigs susceptible to severe heat stress. Our previous studies found that pig small intestine epithelial tissue became significantly damaged following exposure to heat stress, negatively affecting body weight gain. The deleterious effects of heat stress could be ameliorated using a traditional Chinese medicine decoction (CMD), sustaining normal growth while under heat stress. In the current study, we hypothesized the mechanism of CMD activity to be via regulation of gut hormones (NPY, MLN, SCT and GCG) secretion from endocrine cells, which are responsible for nutrient digestion and absorption. To test this, 36 Chinese experimental mini-pigs (2 months of age) were screened according to weight and litter origin, and divided into three treatment groups: control (23 °C for 24 h + standard feed), heat stress (HS; 26 °C for 19 h, 40 °C for 5 h + standard feed) and CMD (26 °C for 19 h, 40 °C for 5 h + standard feed supplemented with CMD); n = 12 per group. Feed intake and body weight gain were measured daily. Pigs were euthanized at days 1 and 6 after initial treatment with blood and sections of small intestine epithelial tissue collected. Serum cortisol (Cor) concentrations were determined using RIA. Endocrine cell number and structural analysis were performed using silver staining, and gut hormone secretion examined by microarray. Dietary supplementation with CMD significantly improved porcine growth performance (P < 0.05), decreased the Cor levels (P < 0.01), increased endocrine cell number as well as up-regulating neuropeptide Y (NPY), motilin (MLN) and secretin (SCT) and down-regulating glucagon (GCG) expression in pig jejunum on day 6 when compared with the HS group. Taken together, our results indicate CMD supplementation can significantly reduce the negative effects of heat stress on pig jejunum, maintaining growth performance similar to non-heat stressed animals. CMD's activity appears to be via adjusting gut hormone secretion to regulate metabolism and improve animal growth.     

Subclinical mastitis causes alterations in nitric oxide, total oxidant and antioxidant capacity in cow milk

The aim of this study was to investigate total antioxidant (TAC), and oxidant capacity (TOC) and nitric oxide (NO) levels in milk of cows with subclinical mastitis. Brown Swiss and Holstein breed cows were screened with California Mastitis Test (CMT) to determine mammary glands with subclinical mastitis. Moreover, somatic cell counts (SCC) were determined electronically in all milk samples. Mammary quarters were classified as healthy (n = 25) or subclinical mastitis (n = 35) based on CMT scores and somatic cell count (SCC: ?200,000/ml or >200,000/ml) in milk. Nitric oxide, TOC and SCC levels were significantly higher (p < 0.001, p < 0.005 and p < 0.001, respectively) in milk from mammary quarters with subclinical mastitis compared to those from healthy mammary quarters. In conclusion, subclinical mastitis results in higher NO concentrations, TOC and SCC, and NO and TOC were positively correlated with SCC. Moreover, alterations in NO levels and TOC in milk could be used as an alternative diagnostic tool to screen for subclinical mastitis.     

Foraging behavior of heifers with or without social models in an unfamiliar site containing high plant diversity

Grazing behavior, diet selection and weight gain of heifers (with or without social models) were assessed in an unfamiliar site containing high plant diversity. The study was performed within a tropical forest ecosystem containing a mixture of grass forb/herb, shrub and tree species. Ten inexperienced crossbred Bos taurus × Bos indicus heifers (7 ± 1 months old) were randomly assigned to one of two groups: naive or naive + social model (sm) and two experienced heifers (16 months old) were assigned to the second group. Naive heifers were bottle-nursed from days 2 to 90 after birth and began grazing at 1 month of age in tropical grass monocultures, while experienced animals had been foraging in sites containing a high diversity of plant species for 3 months prior to the study. Each group grazed in separate paddocks for a 12-week period during the rainy season, and animals were observed using focal sampling from 7:00 to 19:30 h to assess diet composition based on bite counts. Weight gain was assessed every 14 days. Paddocks contained from 1481 to 1789 kg/ha of herbaceous dry matter, enough to support the heifers throughout the study without affecting diet selection. Shrub and tree cover ranged from 55.2 to 58.4% across treatments. Bites per minute were adjusted to a log-scaled quadratic-plateau model and the curves showed no differences between treatments (P = 0.756). However, diet composition differed between groups (P < 0.001), with naive + sm heifers ingesting a greater proportion of trees (P < 0.001) and shrubs (P = 0.02), while naive heifers ingested more forb/herbs (P = 0.02); no difference in grass consumption was observed (P = 0.92). Heifers in both treatments consumed the same plant species (50 from 26 families). Over time, utilization of several plant species increased or decreased (P < 0.05), eventually leveling off for the remainder of the study. Although differences in diet composition were observed, they did not affect overall weight gain (117 and 113 g/day in the naive and naive + sm groups, respectively; P = 0.913). However, initial post-weaning weight loss was avoided in the naive + sm group. Social learning facilitates a higher use of shrubs and trees in tropical pastures containing a high diversity of plant species with different growth habits. As many plant species have high potential as forage, more effort should be placed on developing mechanisms to increase their dietary inclusion by cattle.     

Effects of GnRH treatment on initiation of pulses of LH,LH release,and subsequent concentrations of progesterone

Progesterone is essential for establishment and maintenance of pregnancy. One proposed method to increase progesterone is administering GnRH at insemination. However, this method has resulted in conflicting results. Therefore, 2 experiments were conducted to evaluate how administering GnRH at insemination affected pulses of luteinizing hormone (LH) and subsequent progesterone. In Experiment 1, cows were allotted to 2 treatments: (1) GnRH (100 μg) given approximately 12 h after initiation of estrus (n = 5); and (2) Control (n = 5). Blood samples were collected at 15-min intervals for 6 h at 12 (blood sampling period 1), 26 (blood sampling period 2), 40 (blood sampling period 3), 54 (blood sampling period 4), and 68 (blood sampling period 5) h after onset of estrus. Daily blood samples were collected for 17 d. In Experiment 2, cows were allotted into 2 treatments: GnRH administered 10 to 11 h (n = 10) or 14 to 15 h (n = 10) after onset of estrus. Daily blood samples were collected for 17 d. Cows treated with GnRH tended (P ≤ 0.075) to have greater LH release during blood sampling period 1, tended (P = 0.095) to have fewer pulses during blood sampling period 2, tended (P = 0.067) to have greater concentrations of progesterone, and had an earlier (P = 0.05) increase in progesterone than control cows. Cows treated with GnRH 10 to 11 h after onset of estrus had greater (P = 0.01) progesterone and an earlier (P = 0.04) increase in progesterone than cows treated 14 to 15 h. In conclusion, timing of GnRH treatment following onset of estrus influenced pulses of LH and subsequent progesterone.     

Alteration in lymphocyte responses,cytokine and chemokine profiles in chickens infected with genotype VII and VIII velogenic Newcastle disease virus

Newcastle disease (ND) is a highly contagious avian disease and one of the major causes of economic losses in the poultry industry. The emergence of virulent NDV genotypes and repeated outbreaks of NDV in vaccinated chickens have raised the need for fundamental studies on the virus–host interactions. In this study, the profiles of B and T lymphocytes and macrophages and differential expression of 26 immune-related genes in the spleen of specific-pathogen-free (SPF) chickens, infected with either the velogenic genotype VII NDV strain IBS002 or the genotype VIII NDV strain AF2240, were evaluated. A significant reduction in T lymphocyte population and an increase in the infiltration of IgM⁺ B cells and KUL01⁺ macrophages were detected in the infected spleens at 1, 3 and 4 days post-infection (dpi) (P < 0.05). The gene expression profiles showed an up-regulation of CCLi3, CXCLi1, CXCLi2 (IL-8), IFN-γ, IL-12α, IL-18, IL-1β, IL-6, iNOS, TLR7, MHCI, IL-17F and TNFSF13B (P < 0.05). However, these two genotypes showed different cytokine expression patterns and viral load. IBS002 showed higher viral load than AF2240 in spleen at 3 and 4 dpi and caused a more rapid up-regulation of CXCLi2, IFN-γ, IL-12α, IL-18, IL-1β, iNOS and IL-10 at 3 dpi. Meanwhile, the expression levels of CCLI3, CXCLi1, IFN-γ, IL-12α, IL-1β and iNOS genes were significantly higher in AF2240 at 4 dpi. In addition, the expression levels of IL-10 were significantly higher in the IBS002-infected chickens at 3 and 4 dpi. Hence, infection with velogenic genotype VII and VIII NDV induced different viral load and production of cytokines and chemokines associated with inflammatory reactions.     

Removal of nocturnal secretion of melatonin fails to reduce antibody synthesis and interleukin-2 production of lambs.

Crossbred ewe and wether lambs were used to evaluate the effects of a normal, nocturnal elevation in the concentration of melatonin in the serum on immunological functions. The nocturnal elevation in melatonin was eliminated by exposing half the lambs to constant light (LL), whereas the remainder received a 12-h light, 12-h dark cycle (LD). Immune function was challenged by treating half the lambs in LL and half of the lambs in LD with dexamethasone (DEX; .04 mg/kg); the remainder of the lambs received only a saline vehicle (SAL). The resulting treatment combinations were designated LD+SAL (n = 5), LD+DEX (n = 5), LL+SAL (n = 5), and LL+DEX (n = 5). Lambs were stanchioned individually in environmental rooms; photoperiod treatments commenced on that day (d -14). Also on d -14, lambs were given 1 mg ovalbumin/lamb in adjuvant. Lambs were given a booster injection of .5 mg ovalbumin/lamb on d 0. Treatments with DEX and SAL also began on d 0 and were repeated every 48 h through d 14. Catheters were placed in the jugular vein of all lambs on d 12; samples of plasma and serum were collected hourly from 0800 on d 14 to 0800 on d 15; plasma was assayed for adrenocorticotropic hormone (ACTH) and serum was assayed for cortisol and melatonin. In addition, samples of serum obtained at 0800 on d 15 were used to evaluate antibody titers to ovalbumin. Samples of whole blood also were obtained at 0800 on d 15, and total and differential leukocyte numbers and production of interleukin-2 (IL-2) by lymphocytes were determined.(ABSTRACT TRUNCATED AT 250 WORDS)     

Meta-analysis of the relationship between ractopamine and dietary lysine levels on carcass characteristics in pigs

A meta-analysis was carried out to evaluate the relationship between ractopamine and dietary lysine levels on carcass characteristics in pigs. The database was composed by 29 articles published in international journals from 1990 to 2007, totalizing 155 treatments and 3786 pigs. Average inclusion of ractopamine was 15.3 ppm (ranging from 0 to 30 ppm) and daily average intake of ractopamine was 24.9 mg. Ractopamine addition increased (P < 0.05) hot carcass weight in 4%, loin area in 12% and lean meat content in 4%. Pigs supplemented with ractopamine presented decrease (P < 0.05) of 8% in backfat thickness at the tenth rib, 3% in backfat thickness at the last rib and 5% in mean backfat thickness. Each increase in 1 mg of ractopamine intake represented a reduction of 0.3 mm in tenth-rib (Y = 29.61-0.308 RAC + 0.025 RAC², R² = 0.81, RAC: ractopamine intake expressed in mg) and 0.5 mm at last-rib backfat thickness (Y = 30.52 + 0.519 RAC-0.0054 RAC², R² = 0.94). The use of ractopamine affected (P > 0.05) neither carcass length and dressing, nor meat marbling and color. Loin area was positively correlated (r = 0.27, P < 0.05) and mean backfat thickness was negatively correlated (r = − 0.27, P < 0.05) to dietary lysine concentration. Pigs supplemented with ractopamine whose daily intake of lysine per unit of metabolic weight was more than 195 mg presented (P < 0.05) loin area 4% higher and backfat thickness 10% lower than other animals. Supplemented pigs that received diets with lysine content superior to their calculated amino acid requirement presented weight gain 14% higher, lean meat content 17% higher, leaf fat 34% lower and loin area 6% higher when compared to other supplemented animals. Ractopamine increases lean meat content and reduces backfat thickness in carcass, however, the interaction between additive and nutritional components must be considered in diet formulation.     

Physiopathological effects of zearalenone in post-weaning female piglets with or without montmorillonite clay adsorbent

Zearalenone (ZEA), commonly present in corn and its derived products for animals, has caused significant economical impact on swine reproduction in China. The present study therefore attempted to reveal the adverse effects of ZEA (1.3 mg/kg diet) exposure from a viewpoint of damages focusing on the liver and kidney of female piglets. The efficacy of dietary montmorillonite clay in preventing ZEA-induced adverse effects was also determined. Treatments were 1) control; 2) control + 2.5 g/kg clay; 3) control + 1 mg/kg ZEA; 4) control + 1 mg/kg ZEA + 1.25 g/kg clay; 5) control + 1 mg/kg ZEA + 2.5 g/kg clay; 6) control + 1 mg/kg ZEA + 5.0 g/kg clay; 7) control + 1 mg/kg ZEA + 10 g/kg clay. Results showed that pigs fed ZEA-contaminated diet reduced (P < 0.05) platelets, haemoglobin, globulin, triglycerides and high density lipoproteins (HDL) in serum, and increased (P < 0.05) all enzymes activities, cholesterol, urea, and creatinine. Degeneration of the liver and kidney tissues was also found in female piglets fed 1.3 mg/kg ZEA-contaminated diet. Dietary addition of clay showed a positive protection effect on ZEA feeding, and the linear or quadratic effect (P < 0.05) on neutralizing detrimental effects of clay in ZEA feeding were observed. It suggested that feeding ZEA at 1.3 mg/kg diet for 24-d may result in a deleterious effect in female piglets, and clay addition at 5 or 10 g/kg diet can effectively protect against the detrimental effects of the ZEA feeding. These results may have implications for human and animals consuming ZEA-contaminated food or feed.     

Melatonin receptor subtypes Mel1a and Mel1c but not Mel1b are associated with monochromatic light-induced B-lymphocyte proliferation in broilers

This study determined the effects of melatonin (MEL) and its receptors on monochromatic light-induced bursal B-lymphocyte proliferation in broiler chickens. In vivo, green light (GL) enhanced the proliferation of B lymphocytes in bursas by 16.49% to 30.83% and the expression of MEL receptor subtypes 1a (Mel1a), Mel1b, and Mel1c receptors in bursas by 6.91% to 366.98% than other light colors. However, pinealectomy reduced these parameters and eliminated the differences between GL and other light groups. In vitro, the MEL-induced bursal B-lymphocyte proliferation was most suppressed by prazosin (P = 0.001, selective Mel1c antagonist), followed by luzindole (P = 0.022, nonselective Mel1a/Mel1b antagonist), but not by 4-phenyl-2-propionamideotetralin (P = 0.144, selective Mel1b antagonist). Similarly, dibutyryl-cyclic adenosine monophosphate (cAMP; analog of cAMP; P = 0.017) but not 8-(4-chloro-phenylthio)-2′-O-methyladenosine-3′,5′-cyclic monophosphate (P = 0.736; activator of exchange protein directly activated by cAMP) significantly inhibited bursal B-lymphocyte proliferation. These results suggest that MEL mediates GL-induced bursal B-lymphocyte proliferation through Mel1c and Mel1a receptors but not Mel1b receptors by activating the cAMP/protein kinase A pathway.     

Partial budgeting assessment of the treatment of pyometra, follicular cysts and ovarian inactivity causing postpartum anoestrus in dairy cattle

A total of 412 multiparous German Holstein cows were screened for postpartum pyometra, follicular cysts and ovarian inactivity to assess economic and productivity losses in relation to pharmaceutical expenditures. Our results show that cows treated for pyometra with prostaglandin f2 alpha (PGF2α) and oxytetracycline had significantly (P < 0.05) greater total and net returns than untreated cows or those treated with PGF2α + cephapirin or PGF2α alone. Milk yields from untreated cows affected by follicular cysts were significantly (P < 0.05) lower than the yields from cows treated with gonadotrophin-releasing hormone (GnRH)− and GnRH + PGF2α. In addition, the use of GnRH to treat cows with ovarian inactivity resulted in significantly (P < 0.05) lower costs and greater total and net return values compared to untreated controls.     

Characteristics of prolactin-releasing response to salsolinol (SAL) and thyrotropin-releasing hormone (TRH) in ruminants

The secretion of prolactin (PRL) is stimulated by thyrotropin-releasing hormone (TRH), and inhibited by dopamine (DA). However, we have recently demonstrated that salsolinol (SAL), a DA-derived endogenous compound, is able to stimulate the release of PRL in ruminants. The aims of the present study were to compare the characteristics of the PRL-releasing response to SAL and TRH, and examine the relation between the effects that SAL and DA exert on the secretion of PRL in ruminants in vivo and in vitro. Three consecutive intravenous (i.v.) injections of SAL (5 mg/kg body weight (b.w.): 19.2 μmol/kg b.w.) or TRH (1 μg/kg b.w.: 2.8 nmol/kg b.w.) at 2-h intervals increased plasma PRL levels after each injection in goats (P < 0.05); however, the responses to SAL were different from those to TRH. There were no significant differences in each peak value between the groups. The rate of decrease in PRL levels following the peak was attenuated in SAL-treated compare to TRH-treated animals (P < 0.05). PRL-releasing responses to SAL were similar to those to sulpiride (a DA receptor antagonist, 0.1 mg/kg b.w.: 293.3 nmol/kg b.w.). In cultured bovine anterior pituitary (AP) cells, TRH (10⁻⁸ M) significantly increased the release of PRL following both 15- and 30-min incubation periods (P < 0.05), but SAL (10⁻⁶ M) did not increase the release during the same periods. DA (10⁻⁶ M) completely blocked the TRH-induced release of PRL for a 2-h incubation period in the AP cells (P < 0.05). Sulpiride (10⁻⁶ M) reversed this inhibitory effect but SAL (10⁻⁶ M) did not have any influence on the action of DA. These results show that the mechanism(s) by which SAL releases PRL is different from the mechanism of action of TRH. Furthermore, they also show that the secretion of PRL is under the inhibitory control of DA, and SAL does not antagonize the DA receptor's action.     

Effects of orexigenic peptides and leptin on melatonin secretion during different photoperiods in seasonal breeding ewes: an in vitro study

The pineal gland (PG) acts as a neuroendocrine transducer of daily and seasonal time through the nocturnal release of melatonin. Here, we examined the interaction of season, orexin, ghrelin, and leptin on melatonin secretion by pineal explants in short-term culture. Glands were collected after sunset from 12 ewes during long days (LD; April and May) and from an additional 12 ewes during short days (SD; October and November). Glands were transected sagittally into strips, with each equilibrated in 2.5 mL of Dulbecco's modified Eagle's medium for 60 min, followed by a 2-h incubation in control medium or medium containing orexin B (10 and 100 ng/mL), ghrelin (10 and 100 ng/mL), or 50 ng/mL of leptin. After a 3-h incubation, some PG explants treated previously with lower doses of orexin or ghrelin were challenged with 50 ng/mL of leptin and those treated with both doses of orexin were challenged with 300 nM of the β-agonist isoproterenol. One milliliter of medium was harvested and replaced from each well every 30 min. Treatment with the low dose of orexin during LD increased melatonin secretion about 110% (P<0.01); treatment with a high dose increased melatonin secretion about 47% (P<0.001). During the SD period, leptin stimulated (P < 0.05) melatonin secretion slightly compared with mean melatonin concentration in controls. However, together, orexin and leptin depressed (P<0.01) melatonin secretion. Both doses of ghrelin reduced (P < 0.01) melatonin concentration during the SD season compared with control culture. Addition of ghrelin and leptin to culture medium increased (P<0.01) melatonin concentration compared with ghrelin-treated culture and decreased melatonin concentration (P<0.01) compared with leptin-treated culture during SD. Isoproterenol stimulated (P<0.01) melatonin secretion compared with values observed during the pretreatment period. We conclude that orexigenic peptides (orexin B and ghrelin) and an anorectic peptide (leptin) affect PG directly. The responses of PG to those hormones depend on day length. Moreover, secretion of melatonin from the ovine PG is under an adrenergic regulation.     

Pharmacokinetic and pharmacodynamic interactions of tolfenamic acid and marbofloxacin in goats

Pharmacokinetic and pharmacodynamic properties in goats of the non-steroidal anti-inflammatory drug tolfenamic acid (TA), administered both alone and in combination with the fluoroquinolone marbofloxacin (MB), were established in a tissue cage model of acute inflammation. Both drugs were injected intramuscularly at a dose rate of 2 mg kg⁻¹. After administration of TA alone and TA + MB pharmacokinetic parameters of TA (mean values) were C_max = 1.635 and 1.125 μg ml⁻¹, AUC = 6.451 and 3.967 μg h ml⁻¹, t_1/2K₁₀ = 2.618 and 2.291 h, Vdarea/F = 1.390 and 1.725 L kg⁻¹, and ClB/F = 0.386 and 0.552 L kg⁻¹ h⁻¹, respectively. These differences were not statistically significant. Tolfenamic acid inhibited prostaglandin (PG)E₂ synthesis in vivo in inflammatory exudate by 53-86% for up to 48 h after both TA treatments. Inhibition of synthesis of serum thromboxane (Tx)B₂ ex vivo ranged from 16% to 66% up to 12 h after both TA and TA + MB, with no significant differences between the two treatments.From the pharmacokinetic and eicosanoid inhibition data for TA, pharmacodynamic parameters after dosing with TA alone for serum TxB₂ and exudate PGE₂ expressing efficacy (E_max = 69.4 and 89.7%), potency (IC₅₀ = 0.717 and 0.073 μg ml⁻¹), sensitivity (N = 3.413 and 1.180) and equilibration time (t_1/2K_e0 = 0.702 and 16.52 h), respectively, were determined by PK-PD modeling using an effect compartment model. In this model TA was a preferential inhibitor of COX-2 (COX-1:COX-2 IC₅₀ ratio = 12:1). Tolfenamic acid, both alone and co-administered with MB, did not affect leucocyte numbers in exudate, transudate or blood. Compared to placebo significant attenuation of skin temperature rise over inflamed tissue cages was obtained after administration of TA and TA + MB with no significant differences between the two treatments. Marbofloxacin alone did not significantly affect serum TxB₂ and exudate PGE₂ concentrations or rise in skin temperature over exudate tissue cages. These data provide a basis for the rational use of TA in combination with MB in goat medicine.     

Susceptibility of canine and feline bacterial pathogens to pradofloxacin and comparison with other fluoroquinolones approved for companion animals

In this study, 908 bacterial pathogens from defined infections of dogs and cats were tested for their susceptibility to the novel fluoroquinolone pradofloxacin, which was approved in 2011 for use in cats and dogs. Most of the bacteria tested (Staphylococcus aureus, Staphylococcus pseudintermedius, Escherichia coli, β-haemolytic streptococci, Pasteurella multocida and Bordetella bronchiseptica) exhibited low pradofloxacin MIC₉₀ values of ≤0.25 μg/ml. Solely Proteus spp. and Pseudomonas aeruginosa had higher MIC₉₀ values of ≥4 μg/ml. Only six (3.4%) of 177 S. pseudintermedius and 12 (5.3%) of 227 E. coli isolates showed pradofloxacin MICs of ≥2 μg/ml. Analysis of the quinolone resistance determining regions of the target genes identified double mutations in GyrA that resulted in amino acid exchanges S83L + D87N or S83L + D87Y and single or double mutations in ParC that resulted in amino acid exchanges S80I or S80I + E84G in all 12 E. coli isolates. The six S. pseudintermedius isolates exhibited amino acid exchanges S84L or E88K in GyrA and S80I in GrlA. Comparative analysis of the MICs of pradofloxacin and the MICs determined for enrofloxacin and its main metabolite ciprofloxacin, but also marbofloxacin, orbifloxacin, difloxacin and ibafloxacin was conducted for the target pathogens S. pseudintermedius, E. coli and P. multocida. This comparison confirmed that pradofloxacin MICs were significantly lower than those of the other tested fluoroquinolones.