Magnetic hydrogel particles improve nanopore sequencing of SARS-CoV-2 and other respiratory viruses.

Presented here is a magnetic hydrogel particle enabled workflow for capturing and concentrating SARS-CoV-2 from diagnostic remnant swab samples that significantly improves sequencing results using the Oxford Nanopore Technologies MinION sequencing platform. Our approach utilizes a novel affinity-based magnetic hydrogel particle, circumventing low input sample volumes and allowing for both rapid manual and automated high throughput workflows that are compatible with Nanopore sequencing. This approach enhances standard RNA extraction protocols, providing up to 40 × improvements in viral mapped reads, and improves sequencing coverage by 20-80% from lower titer diagnostic remnant samples. Furthermore, we demonstrate that this approach works for contrived influenza virus and respiratory syncytial virus samples, suggesting that it can be used to identify and improve sequencing results of multiple viruses in VTM samples. These methods can be performed manually or on a KingFisher automation platform.

Hydrogel particles improve detection of SARS-CoV-2 RNA from multiple sample types.

Here we present a rapid and versatile method for capturing and concentrating SARS-CoV-2 from contrived transport medium and saliva samples using affinity-capture magnetic hydrogel particles. We demonstrate that the method concentrates virus from 1 mL samples prior to RNA extraction, substantially improving detection of virus using real-time RT-PCR across a range of viral titers (100-1,000,000 viral copies/mL) and enabling detection of virus using the 2019 nCoV CDC EUA Kit down to 100 viral copies/mL. This method is compatible with commercially available nucleic acid extraction kits (i.e., from Qiagen) and a simple heat and detergent method that extracts viral RNA directly off the particle, allowing a sample processing time of 10 min. We furthermore tested our method in transport medium diagnostic remnant samples that previously had been tested for SARS-CoV-2, showing that our method not only correctly identified all positive samples but also substantially improved detection of the virus in low viral load samples. The average improvement in cycle threshold value across all viral titers tested was 3.1. Finally, we illustrate that our method could potentially be used to enable pooled testing, as we observed considerable improvement in the detection of SARS-CoV-2 RNA from sample volumes of up to 10 mL.

Preliminary comparative serological evaluation of Histamine H2 receptors in dogs with an acute onset of vomiting treated with Ranitidine and healthy dogs.

Despite the lack of evidence of the ability to suppress gastric acid secretion in dogs, ranitidine (RT) is often used to control clinical signs in dogs with acute vomiting even if the way it happens it is still largely unknown. The aim of this study was thus to evaluate the therapeutic effect of ranitidine on H2 receptors in dogs with acute vomiting. To investigate the RT activity a preliminary study was performed in dogs which underwent gastroscopy analyses, demonstrating that the level of H2R observed in the serum and gastric wall tissue was the same [1.473(1.30; 1.79) ng/ml and 1.498 (1.33; 1.85) ng/ml, respectively]. After that H2R levels in the serum of 22 healthy dogs (Group 1) and in a group of 22 dogs with acute vomiting (Group 2) were compared both before (T0), after 7-10 days (T1) of 2 mg/kg twice a day ranitidine administration and after 11 days since the drug was discontinued (T2). Significant differences (p<0.001) were detected between the level of circulating H2R among Group 1: 0.41 ng/ml (0.28;0.54) and Group 2: 2.27 ng/ml (2.11;2.49) at T0. In Group 2, no difference in the level of H2R was detected in samples collected at T0 compared to those at T1 [T1: 2.32 ng/ml (2.14; 2.49)] and T2 [T2: 2.30 ng/ml (1.99;2.69)]. In Group 2 all patients but one displayed remission of symptoms attributable to inflammatory gastropathy at the first withdrawal (T1: 7-10 days), while at the second withdrawal (T2: after 21 days), remission was detected in all dogs. Our preliminary hypothesis is that the clinical efficacy of ranitidine is related to the greater expression of H2 receptors in patients with acute vomiting. This increased expression may be due to continuous pathological stimulus at the gastric level. Further studies with a wider population are needed to better investigate the activity of RT in dogs with acute onset of vomiting.

Análise bioeconômica de períodos de suplementação proteico-energética na estação seca para novilhas Nelore em pastagem diferida de Urochloa decumbens / Bioeconomic analysis of protein-energy supplementation periods in dry season for Nellore heifers in stockpiled pastures of Urochloa decumbens

A suplementação proteico-energética (SPE) é uma estratégia adequada à bovinocultura de corte no período seco do ano, embora sua adoção equivocada possa comprometer seu benefício/custo. O objetivo deste estudo foi avaliar o desempenho de novilhas Nelore em pastagem diferida de Urochloa decumbens sob dois períodos de SPE, e realizar análise econômica desses sistemas de produção. O experimento foi conduzido sob delineamento inteiramente ao acaso, com 147 e 55 dias de SPE (sistemas 1 e 2, respectivamente), de 21/06 a 15/11/2016, com 16 e 20 novilhas nos sistemas 1 e 2, respectivamente. Os dados de desempenho foram analisados utilizando-se o PROC MIXED do SAS®, e as médias comparadas por PDIFF (P<0,05). Os pesos médios foram semelhantes (P>0,05) entre os sistemas de produção nas pesagens de 21/06, 12/07, 02/08, 23/08 e 15/11/2016. Os valores presentes líquidos foram positivos sob a taxa anual de desconto de 4%, sendo viáveis financeiramente. Por apresentarem taxa interna de retorno superior à taxa mínima de atratividade, os sistemas foram atrativos para investimento. Foi obtido balanço econômico positivo de R$ 25,26/animal para substituição do sistema 1 pelo sistema 2. A redução de 92 dias na SPE proporciona desempenho semelhante entre novilhas e maior eficiência econômica do sistema de produção.(AU)

Protein-energy supplementation (SPE) is an adequate strategy for beef cattle during drought season of the year, although the wrong adoption might compromise its cost/benefit ratio. The aim of this study was to evaluate Nellore heifers' performance under SPE periods and to draw economic analysis of production systems. The experiment was conducted under a completely randomized design, with two systems: 147 and 55 days of SPE (Systems 1 and 2), from 06/21/2016 to 11/15/2016, with 16 and 20 heifers to Systems 1 and 2, respectively. Performance data were analyzed by SAS® PROC MIXED and means compared by PDIFF (P< 0.05). The weight means were similar between the production systems (P> 0.05) at weighing of 06/21, 07/12, 08/02, 08/23 and 11/15/2016. The net present values were positive under annual discount rate of 4%, being financially viable. By presenting internal return rate higher than the minimum attractiveness rate, the systems were attractive for investment. Positive economic balance was obtained of R$ 25.26/animal to replace System 1 by 2. The reduction of 92 days in SPE provides similar performance between Nellore heifers and greater economic efficiency of the production system.(AU)

Effect of forage species and supplement type on rumen kinetics and serum metabolites in growing beef heifers grazing winter forage.

The objective of this study was to determine the effect of stockpiled forage type and protein supplementation on VFA production, serum metabolites, and BW in yearling beef heifers. Over 2 yr, spring-born, Angus crossbred yearling beef heifers ( = 42; 305 ± 2.9 kg initial BW) were randomly assigned to 1 of 3 forage pasture types: 1) endophyte-infected tall fescue [TF; (Schreb.) Dumort], 2) a big bluestem ( Vitman) and indiangrass ( L.) combination (BI), or 3) switchgrass (SG; L.). Each pasture was then randomly assigned to receive either 1 of 2 isonitrogenous CP treatments: 1) 0.68 kg·heifer·d of dried distiller's grains with solubles (DDGS; 28% CP and 88% TDN) or 2) 0.22 kg·heifer·d of blood meal and fish meal (BF; 72.5% CP and 69.5% TDN), resulting in a 3 × 2 factorial arrangement of treatments. Treatments were initiated in January and terminated in April in both years of the study. Body weights and blood samples were collected approximately every 28 d from initiation of grazing until the end of the trial. Heifer BW change from January to February and overall BW change were greater ( < 0.01) for TF heifers. However, BW change from March to April was not different ( = 0.84) among forage types. Supplement type did not influence ( ≥ 0.13) BW or BW change from January to February and from January to April; however, heifers fed DDGS had greater ( = 0.03) BW gain from March to April. Heifer BW change from February to March exhibited ( < 0.05) a forage type × supplement interaction, with BF-fed heifers gaining more BW on BI pastures than DDGS-fed heifers. Serum glucose concentrations, ruminal acetate, and the acetate:propionate ratio were greater ( ≤ 0.04) for SG heifers. However, circulating serum NEFA and urea N (SUN) concentrations were not different ( ≥ 0.85) among forage types. Serum glucose and NEFA concentrations were not influenced ( ≥ 0.61) by supplement type. Circulating SUN concentrations were greater ( < 0.01) in BF-supplemented heifers. Ruminal acetate tended to be greater ( = 0.09) and butyrate concentrations were greater ( < 0.01) for BF-supplemented heifers. The acetate:propionate ratio was not influenced ( = 0.15) by supplement type. These results suggest that a compensatory gain period prior to breeding would be needed for these native warm-season species to be a viable opportunity for growing and developing replacement heifers in the southeastern United States.

Rabbit dietary supplementation with pale purple coneflower. 1. Effects on the reproductive performance and immune parameters of does.

Echinacea pallida (EPAL), also known as pale purple coneflower, is a herbaceous flowering plant with immune-enhancement and antioxidative properties. The effect of EPAL on the reproductive performance, serum biochemistry and haematological parameters of rabbit does has been studied here. A total of 100, 21-week-old Grimaud rabbit does, were randomly assigned to two groups. One group was fed a basal diet supplemented with 3 g EPAL/kg diet (Echinacea group, E), while the other was fed the basal diet without the supplementation (control group, C). The reproductive performance of the does was not affected by the treatment (P>0.05). The haematological parameters of pregnant rabbits showed that there was no interaction between gestation day and treatment. The EPAL supplementation induced a reduction (-47.3%) in the basophil cell rate (0.55% and 0.29%, for the control and treatment groups, respectively; P=0.049). The gestation day significantly affected most of the haematological parameters (P<0.05). The white blood cell counts declined progressively after day 14. The mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red cell distribution width, mean platelet volume and eosinophils increased steadily throughout the study, and reached a maximum value on day 28. The red blood cells, haemoglobin, haematocrit, mean corpuscular volume and neutrophils increased slightly up to day 14, and then subsequently decreased progressively until day 28. The lymphocytes and platelet distribution width decreased until day 14, and then increased to a maximum value on day 28. No significant effect of gestation day or treatment was observed on the blood serum chemistry. As far as the immune parameters are concerned, no significant differences were observed between groups, while a significant effect of gestation day was observed for lysozymes (6.02 v. 7.99 v. 1.91; for 0, 14 and 28 days, respectively; P=0.014). In conclusion, a lack of effect of EPAL has been observed. In fact, no impacts of EPAL have been observed on the reproductive or haematological parameters of the does. The effects of dietary supplementation with EPAL on the performances, bacterial community, blood parameters and immunity in growing rabbits are reported in the second part of this study.

Rabbit dietary supplementation with pale purple coneflower. 2. Effects on the performances, bacterial community, blood parameters and immunity of growing rabbits.

Echinacea pallida (EPAL), a herbaceous flowering plant with immunomodulatory properties, has been chosen to determine the pre- and post-supplementary effects on the growth performances, bacterial community, blood parameters and immunity of growing rabbits. The same Grimaud does (14-week-old) from the studied in the first part of this study were randomly divided into two groups (n=50/group). The first group was fed a basal diet without supplementation (Control group, C) while the another group was fed a basal diet supplemented with 3 g EPAL/kg diet (Echinacea group, E). From the second parturition, 80 weaned kits (40 from the C does and 40 from the E does) were randomly assigned to four groups of 20 animals each and were fed a growing commercial diet supplemented with or without a 3 g EPAL/kg diet: the CC group (rabbits from the C does fed the control diet), CE group (rabbits from the C does fed the supplemented diet), EC (rabbits from the E does fed the control diet) and EE group (rabbits from the E does fed the supplemented diet). The dietary EPAL treatment did not affect the growth performance. Ten fattening rabbits from each group were selected to evaluate the bacterial community and blood parameters, while the remaining rabbits (n=10/group) were used to study phagocytosis and the humoral immune response. The variability was evaluated from hard faeces at 35, 49 and 89 days, and the caecal content at 89 days. The variability of the bacterial community of the EE group was higher than that of the other groups. The phagocytic activity was higher in the CE and EE groups than in the CC and EC ones (30.9 and 29.7 v. 21.2 and 21.8%; P<0.05), whereas no statistically significant difference was observed for the blood parameters or humoral immune response against vaccination (rabbit haemorrhagic disease virus) at 95 days old which the serum was collected at 88, 102, 109, 116 and 123 days old. In conclusion, no impact of EPAL dietary supplementation has been observed on the growth performances, bacterial community, blood parameters or humoral immune responses in growing rabbits, except for an increase in phagocytic activities.

Botulinum toxin type A as an adjunct in postoperative pain management in dogs undergoing radical mastectomy.

The aim of this randomised placebo-controlled, observer-blinded study was to evaluate the analgesic effects of botulinum toxin type A (BoNT-A) as an adjunct for postoperative pain control in dogs. Sixteen dogs undergoing bilateral radical mastectomy for treatment of mammary tumours were enrolled. Twenty-four hours before surgery, the subjects were distributed into two groups of eight dogs each: 7 iu/kg BoNT-A (BoNT-A) or saline (Control) was administered subcutaneously in each mammary gland. Following sedation with intramuscular 0.03 mg/kg acepromazine and 0.3 mg/kg morphine, anaesthesia was induced intravenously with 4 mg/kg propofol and maintained with isoflurane/O2. Postoperative analgesia was evaluated for 72 hours after extubation using the Visual Analogue Scale (VAS) and modified Glasgow Composite Measure Pain Scale (modified-GCMPS). Rescue analgesia was provided with intramuscular morphine (0.5 mg/kg). Data were analysed using analysis of variance, Tukey's test, Mann-Whitney U test and Friedman test (P<0.05). The pain scores were significantly lower in the BoNT-A than in the Control from 8 hours to 60 hours and from 12 hours to 60 hours after extubation, based on the VAS and modified-GCMPS, respectively. Rescue analgesia was required by significantly more dogs in the Control (7/8) compared with the BoNT-A (2/8) (P=0.022). Pre-emptive BoNT-A appears to be effective as an adjuvant for postoperative pain management in dogs undergoing bilateral radical mastectomy.

Expression and functionality of TRPV1 receptor in human MCF-7 and canine CF.41 cells.

As canine mammary tumours (CMT) and human breast cancer share clinical and prognostic features, the former have been proposed as a model to study carcinogenesis and improved therapeutic treatment in human breast cancer. In recent years, it has been shown that transient receptor potential vanilloid 1 (TRPV1) is expressed in different neoplastic tissues and its activation has been associated with regulation of cancer growth and progression. The aim of the present research was to demonstrate the presence of TRPV1 in human and canine mammary cancer cells, MCF-7 and CF.41, respectively, and to study the role of TRPV1 in regulating cell proliferation. The images obtained by Western blot showed a signal at 100 kDa corresponding to the molecular weight of TRPV1 receptor. All tested TRPV1 agonists and antagonists caused a significant decrease (P < 0.05) of cell growth rate in MCF-7 cells. By contrast, in CF.41 cells capsaicin and capsazepine induced a significant increase (P < 0.05) in cell proliferation, whereas resiniferatoxin (RTX) and 5-iodo-resiniferatoxin (5-I-RTX) had no influence on CF.41 cell proliferation. Further studies are needed to elucidate the underlying molecular mechanism responsible for the different effects evoked by TRPV1 activation in MCF-7 and CF.41 cells.

Desempenho de novilhos de corte em pastos de capim-tanzânia sob quatro alturas de desfolha / Performance of steers on pasture of Tanzania grass under four defoliation heights

Determinou-se a melhor altura de pastejo do capim-tanzânia (Panicum maximum Jacq. cv. Tanzânia) sob lotação contínua e com taxa de lotação variável na produção de bovinos de corte. A área experimental, de 12 hectares, foi dividida em piquetes de um hectare cada, com três repetições para cada altura - 20; 40; 60 e 80cm. Foram utilizados, em cada piquete, três bovinos machos, da raça Nelore, mais animais adicionais conforme necessidade de ajuste à altura pretendida. O delineamento experimental foi inteiramente ao acaso. A taxa de lotação apresentou comportamento linear negativo em razão das alturas de pastejo em todas as estações do ano, e o mesmo efeito foi observado com o ganho de peso vivo por hectare, enquanto o ganho médio diário apresentou comportamento contrário. Alturas de capim-tanzânia entre 40 e 60cm, sob lotação contínua, possibilitam bom ganho de peso médio diário em bovinos de corte e boa produção por hectare...

This study was carried out to evaluate the effects of four grazing heights under continuous stocking of Tanzânia grass (Panicum maximum Jacq. Cv. Tanzânia) on the stocking rate, performance and beef production per hectare. The experimental area was 12 hectares, divided into paddocks with one hectare each, with three replicates each (20, 40, 60 and 80 cm). There were three male Nellore bovines per paddock, adding animals as needed to adjust the desired heights in a completely randomized design. The stocking rate had a negative linear function of grazing heights in all seasons regarding yield per hectare, while the average daily gain showed opposite behavior. Apparently, Tanzânia grass heights between 40 and 60cm under continuous stocking enable good average daily weight gain in beef cattle, and high yield per hectare...

In vitro enantioselective pharmacodynamics of Carprofen and Flunixin-meglumine in feedlot cattle.

The activity of the anti-inflammatory agents Flunixin-meglumine (FLU), RS (±) Carprofen (CPF) and S (+) CPF on bovine cyclooxygenases (COXs) has been characterized in feedlot calves using an in vitro whole blood model. The drugs showed equivalent efficacy in their inhibitory activity on COXs, and the rank order of potency for both COX-1 and COX-2 inhibition was FLU > S (+) CPF > RS (±) CPF. Our results indicated that FLU is a nonselective inhibitor of bovine COXs, whereas RS (±) CPF and S (+) CPF exhibited different degrees of preferential inhibition of COX-2 isoenzyme. The rank order of IC50 COX-1: IC50 COX-2 potency ratios was in fact S (+) CPF (51.882) > RS (±) CPF (13.964) > FLU (0.606), and the calculated percentage inhibition of COX-1 corresponding to COX-2 inhibition values comprised between 80% and 95% was comprised between 57.697 and 79.865 for FLU, 33.373 and 51.319 for RS (±) CPF, and 0.230 and 4.622 for S (+) CPF, respectively. These findings are discussed in relation to the prediction of the clinical relevance of COX inhibition by the test drugs in cattle.

Effects of flunixin meglumine and ketoprofen on mediator production in ex vivo and in vitro models of inflammation in healthy dairy cows.

In this study, ex vivo assays were carried out in dairy cows to evaluate the anti-inflammatory effects of two nonsteroidal anti-inflammatory drugs: ketoprofen (KETO) and flunixin meglumine (FM). Twelve healthy Holstein dairy cattle were randomly allocated to two groups (n=6): group 1 received FM and group 2 received KETO at recommended therapeutic dosages. The anti-inflammatory effects of both drugs were determined by measuring the production of coagulation-induced thromboxane B2 (TXB2 ), lipopolysaccharides (LPS) (10 µg/mL)-induced prostaglandin E2 (PGE2 ), and calcium ionophore (60 µm)-induced leukotrien B4 (LTB4 ). Cytokine production was assessed by measuring tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and interleukin-8 (CXCL8) concentrations after incubation in the presence of 10 µg/mL LPS. The IC50 of FM and KETO was determined in vitro by determining the concentration of TXB2 and PGE2 in the presence of scalar drug concentrations (10(-9) -10(-3) m). Both FM and KETO inhibited the two COX isoforms in vitro, but showed a preference for COX-1. FM and KETO showed similar anti-inflammatory effects in the cow.

In vitro and ex vivo pharmacodynamics of selected non-steroidal anti-inflammatory drugs in equine whole blood.

Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit cyclooxygenases (COX), and the inhibition of COX-2 rather than COX-1 can limit the onset of NSAID-related adverse effects. The pharmacodynamic properties of eltenac, naproxen, tepoxalin, SC-560 and NS 398 in healthy horses were investigated using an in vitro whole blood assay. To predict COX selectivity in clinical use, eltenac and naproxen were also studied ex vivo after intravenous administration. SC-560 acted as a selective COX-1 inhibitor, tepoxalin as a dual inhibitor with potent activity against COX-1, and NS 398 as a preferential COX-2 inhibitor. Eltenac was a preferential COX-2 inhibitor in vitro but un-selective in the ex vivo study. Naproxen maintained its non-selectivity both in vitro and ex vivo. These findings have demonstrated that in vitro studies may not accurately predict in vivo NSAID selectivity for COX and should be confirmed using an ex vivo whole blood assay.

Use of self-actuating and self-sensing cantilevers for imaging biological samples in fluid.

In this paper, we present a detailed investigation into the suitability of atomic force microscopy (AFM) cantilevers with integrated deflection sensor and micro-actuator for imaging of soft biological samples in fluid. The Si cantilevers are actuated using a micro-heater at the bottom end of the cantilever. Sensing is achieved through p-doped resistors connected in a Wheatstone bridge. We investigated the influence of the water on the cantilever dynamics, the actuation and the sensing mechanisms, as well as the crosstalk between sensing and actuation. Successful imaging of yeast cells in water using the integrated sensor and actuator shows the potential of the combination of this actuation and sensing method. This constitutes a major step towards the automation and miniaturization required to establish AFM in routine biomedical diagnostics and in vivo applications.

Palmitoylethanolamide, endocannabinoids and related cannabimimetic compounds in protection against tissue inflammation and pain: potential use in companion animals.

Endocannabinoids have analgesic/anti-inflammatory properties. The biology of endocannabinoids, their receptors, signalling mechanisms and role in the regulation of physiological processes have been extensively reviewed. This review focuses on the role of palmitoylethanolamide (PEA), an endogenous fatty acid amide analogue of the endocannabinoid anandamide, in tissue protective mechanisms. PEA was first identified almost five decades ago in lipid extracts of various natural products, and its anti-inflammatory and antinociceptive effects were established later. Evidence exists that PEA is synthesised during inflammation and tissue damage and a number of beneficial effects, including the relief of inflammation and pruritus, have been shown to be useful in the control of neurogenic and neuropathic pain. The postulated hypotheses as to the mode of action of PEA include a possible local autacoid-like mediator activity regulating mast-cell activity and putative activation of cannabinoids and vanilloid TRPV1 receptors via "entourage" effects. The large number of scientific investigations into the effects of PEA and PEA-related compounds has given rise to new therapeutic opportunities. In spite of the multitude of therapies currently employed to control inflammation, pain, pruritus and tissue damage, the possibility of using a natural compound, such as PEA to manipulate endogenous protective mechanisms may be considered a beneficial novel therapeutic strategy in veterinary medicine.

Mepartricin long-term administration regulates steroid hormone and adrenergic receptor concentrations in the prostate of aged rats.

Mepartricin is a semi-synthetic macrolide antibiotic developed as a drug for the treatment of benign prostatic hyperplasia (BPH) in human patients. In the present study, aged rats are used as an experimental model to evaluate the effects of mepartricin on circulating hormone concentrations and prostate receptor concentrations, to compare these possible effects with clinical findings observed in long-term treated dogs. Fifty-six aged male rats were randomly divided into four experimental groups treated orally with 0 (group 1), 2 mg (group 2), 5 mg (group 3) and 20 mg (group 4) mepartricin/kg of body weight. for 28 days respectively. Serum oestradiol and testosterone concentrations were measured by radio-immune-assays methods. Binding assays were used to measure the prostate concentrations of oestrogen receptors (ER), androgen receptors (AnR), alpha(1)-adrenergic receptor (alpha(1)-AR), and beta-adrenerergic receptor (beta-AR) subtypes. Mepartricin induced a significant reduction of prostate weight and serum oestradiol concentrations. Serum testosterone concentrations were unaffected. The treatment induced a significant down-regulation of ER concentrations (P < 0.05) and a significant up-regulation of AnR (P < 0.05) in rat prostate. Mepartricin induced a significant (P < 0.05) dose-dependent up-regulation of alpha(1)-AR and beta(2)-AR. In contrast, the concentration of beta(3)-ARs was significantly decreased (P < 0.05) in treated animals. The increase in prostate beta(2)-AR concentrations observed in subjects treated with mepartricin may be a favourable element in the evolution of BPH, because of the role exerted by these receptors in the control of prostatic smooth muscle relaxation. Curiously, beta(3)-AR concentrations were significantly reduced in treated animals. Data collected suggest that the prostatic beta-AR expression might be strongly influenced by oestrogen deprivation (mepartricin treatment); therefore, the combination of oestrogen suppression (mepartricin) and adrenergic suppression (alpha(1)-AR blockers) may be proposed as a possible nonhormonal therapeutic strategy for the treatment of benign prostatic hyperplasia in dogs.

Changes in lymphocyte glucocorticoid and beta-adrenergic receptors in veal calves treated with clenbuterol and steroid hormones for growth-promoting purposes.

In order to identify possible peripheral markers of illegal treatments with growth-promoting agents in veal calves, beta-adrenergic receptor (beta-AR) and glucocorticoid receptor (GR) concentrations were measured in lymphocytes of 12 male Friesian crossbred calves (six controls and six treated). The animals received a cocktail of anabolic and re-partitioning agents [17beta-oestradiol: 3 x 10 mg intramuscular (i.m.) doses at 17-day intervals; dexamethasone sodium phosphate: 4 mg/day for 6 days and 5 mg/day for six further days dissolved in milk; and clenbuterol: 20 microg/kg/day dissolved in milk for the last 40 days before slaughter]. Blood samples were collected by venipuncture at different time points and lymphocytes were isolated by density gradient centrifugation. Lymphocyte beta-AR and GR levels were measured by binding assays. Treatment with re-partitioning agents caused a significant down-regulation of lymphocyte beta-ARs 19 days after the beginning of clenbuterol administration and at day 55 (after dexamethasone withdrawal, just before slaughter). This phenomenon was partially reversed at day 50, after dexamethasone administration, at which time a significant decrease in GR concentrations also occurred. For both types of receptors, no significant changes in the dissociation constant values were observed at any time point. Lymphocytes express measurable concentrations of beta-ARs and GRs and the measurement of receptor levels highlights the fluctuation of receptor expression due to the dynamic interaction of the drugs used in combination. Lymphocyte receptor determination could therefore be included in a battery of biological assays to detect illegal treatments with anabolic agents in veal calves in the light of a multivariate approach.

Measurement of faecal corticoid metabolites in domestic dogs.

In the present study we established a method for the determination of faecal glucocorticoid metabolites in dogs and then used the assay to evaluate the adrenocortical activity in 12 dogs divided into two groups. In group A faecal samples were collected at their domestic setting. In group B, faecal samples were collected at home prior to transport to a boarding kennel, where faecal samples were then collected. In faecal samples most of the steroids were extracted with methanol and determined using a radioimmunoassay with an anti-cortisol antibody. Dogs in group A did not show any statistically significant inter-day variations in the basal levels of faecal corticoid metabolites. Faecal corticoid metabolites in dogs in group B were significantly higher on the first day at the kennel compared to animals kept at home. The peak concentration was found after 24 hours and followed by a slow decline. These results suggest that extraction with methanol and dosage with an anti-cortisol antibody by radioimmunoassay represents a valid approach technique for determination of faecal glucocorticoid metabolites and accurately reflects adrenocortical activity.