Clinical efficacy of bronchodilators in equine asthma: Looking for minimal important difference.

BACKGROUND: Airway obstruction is the main trait of severe equine asthma that affects respiratory function and elicits detrimental effects on clinical presentation. Only few and underpowered clinical studies have investigated the impact of improvement in lung function induced by bronchodilators on the clinical signs of asthma-affected horses. OBJECTIVES: To identify the minimal important difference (MID) in lung function elicited by bronchodilator leading to a meaningful improvement in clinical signs. STUDY DESIGN: Pairwise meta-analysis and meta-regression analysis. METHODS: Literature searches were performed for studies that investigated the effect of bronchodilator therapy on lung function and clinical condition of asthmatic horses. The relationship between the change in lung function variables and clinical score was analysed via random-effect meta-regression. One-point change of the Improved clinically Detectable Equine Asthma Scoring System (IDEASS) score was used to identify the MID. RESULTS: A significant (P<0.05) relationship was found between the changes in IDEASS score and maximum change in transpulmonary pressure (ΔPplmax ) or pulmonary resistance (RL ). Since only the model resulting for RL passed through the origin (Y-intercept when X = 0: -0.31, 95% CI -0.75 to 0.14), this variable was used to identify the MID correlated with a meaningful improvement in clinical signs. The resulting MID value was a change in RL of 0.63 cm H2 O/L/s (95% CI 0.33-0.94), representing the slope of meta-regression model (high quality of evidence). MAIN LIMITATIONS: No long-term studies investigated the effect of bronchodilator agents on both lung function and clinical signs in asthmatic horses. CONCLUSIONS: In conclusion, bronchodilator pharmacotherapy in equine asthma elicits clinically meaningful effect when RL increases ≥1 cm H2 O/L/s, a value indicating the MID. Assessing the MID based on change in RL may improve the quality of evidence and the scientific impact of future clinical trials as it extends beyond the simple, and limiting, evaluation of statistical significance.

Geographical characteristics influencing the risk of poisoning in pet dogs: Results of a large population-based epidemiological study in Italy.

Pets can act as sentinels for human health and thus surveillance of pet dogs has the potential to improve awareness of emerging risks for animal and public health. The aim of this study was to investigate factors associated with the risk of canine poisoning. In a large population-based epidemiological investigation in Italy performed from January 2015 to January 2016 and April 2016 to April 2017, descriptive statistics were acquired and analysed to determine variables associated with poisoning events in pet dogs. Results were validated in a test population and forecast analysis of risk was performed. The cumulative incidence of poisoning events was low (10.2/1000 dogs/year). Anticoagulant rodenticides, organophosphate pesticides, metaldehyde and strychnine were the most frequent causes of intoxications. Territory characteristics significantly modulated both the frequency and the nature of the involved substances. The seashore area was associated with poisoning by rodenticides (odds ratio, OR, 1.81, 95% confidence interval, CI, 1.54-2.13) and metaldehyde (OR 1.61, 95% CI 1.16-2.28). The hill country area was associated with poisoning by organophosphate pesticides (OR 1.73, 95% CI 1.38-2.15), metaldehyde (OR 2.26, 95% CI 1.53-3.25) and strychnine (OR 1.86, 95% CI, 1.34-2.57). The mountain area was associated with strychnine poisoning (OR 3.79, 95% CI 2.84-5.06). The prospective cumulative incidence of poisoning over 10 years was 9.74% (95% CI 9.57-9.91). These results may be useful for predicting the risk of poisoning and for estimating the risk index related to specific toxic compounds in specific territories. This study suggests that poisoning events in dogs may represent a problem of public health with the potential to affect wildlife and human beings.

Clinical effect of corticosteroids in asthma-affected horses: A quantitative synthesis.

BACKGROUND: There are limited findings from low-powered studies based on few number of subjects with equine asthma. Furthermore, no studies have been performed to assess a meaningful clinically detectable impact of corticosteroids in equine asthma. OBJECTIVES: To assess and compare the clinical effect of inhaled and systemic corticosteroids in equine asthma and identify a quantitative clinical score suitable to assess the Minimal Important Difference (MID), expressed as the Minimally Clinically Detectable Difference (MCDD). STUDY DESIGN: Pair-wise and network meta-analysis. METHODS: Literature searches for studies on corticosteroid therapy in equine asthma were performed. The risk of publication bias was assessed by Funnel plots and Egger's test. The effect on changes in clinical scores vs. control was analysed via random-effects models and Bayesian networks. RESULTS: Corticosteroids significantly improved the clinical condition (Standardised Mean Difference: -1.52, 95% CrI -2.07 to -0.98; P<0.001 vs. control). No difference was detected between inhaled and systemic corticosteroids with regard to the changes in clinical scores (Relative Effect: 0.08, 95% CrI -1.45 to 1.32; P = 0.8). An Improved clinically Detectable Equine Asthma Scoring System (IDEASS) indicated that corticosteroids improved the clinical condition of asthmatic horses by 30% compared with controls (IDEASS value: -2.36, 95% CI -3.39 to -1.33; P<0.001). A one-point change in IDEASS represented the MCDD in equine asthma. MAIN LIMITATIONS: Moderate quality of evidence for systemic corticosteroids. CONCLUSIONS: Inhaled corticosteroids are effective in improving the clinical condition of horses with equine asthma and prevent exacerbations. Systemic corticosteroids should be used only in selected cases with symptomatic airway hyperresponsiveness during exacerbation. IDEASS requires further validation but may represent a suitable approach to rank the level of asthma severity and assess the clinical effect of pharmacotherapy in horses with equine asthma.

Pharmacological treatments in asthma-affected horses: A pair-wise and network meta-analysis.

BACKGROUND: Equine asthma is a disease characterised by reversible airflow obstruction, bronchial hyper-responsiveness and airway inflammation following exposure of susceptible horses to specific airborne agents. Although clinical remission can be achieved in a low-airborne dust environment, repeated exacerbations may lead to irreversible airway remodelling. The available data on the pharmacotherapy of equine asthma result from several small studies, and no head-to-head clinical trials have been conducted among the available medications. OBJECTIVES: To assess the impact of the pharmacological interventions in equine asthma and compare the effect of different classes of drugs on lung function. STUDY DESIGN: Pair-wise and network meta-analysis. METHODS: Literature searches for clinical trials on the pharmacotherapy of equine asthma were performed. The risk of publication bias was assessed by funnel plots and Egger's test. Changes in maximum transpulmonary or pleural pressure, pulmonary resistance and dynamic lung compliance vs. control were analysed via random-effects models and Bayesian networks. RESULTS: The results obtained from 319 equine asthma-affected horses were extracted from 32 studies. Bronchodilators, corticosteroids and chromones improved maximum transpulmonary or pleural pressure (range: -8.0 to -21.4 cmH2 O; P<0.001). Bronchodilators, corticosteroids and furosemide reduced pulmonary resistance (range: -1.2 to -1.9 cmH2 O/L/s; P<0.001), and weakly increased dynamic lung compliance. Inhaled ß2 -adrenoreceptor (ß2 -AR) agonists and inhaled corticosteroids had the highest probability of being the best therapies. Long-term treatments were more effective than short-term treatments. MAIN LIMITATIONS: Weak publication bias was detected. CONCLUSIONS: This study demonstrates that long-term treatments with inhaled corticosteroids and long-acting ß2 -AR agonists may represent the first choice for treating equine asthma. Further high quality clinical trials are needed to clarify whether inhaled bronchodilators should be preferred to inhaled corticosteroids or vice versa, and to investigate the potential superiority of combination therapy in equine asthma.

Proteomics and the search for welfare and stress biomarkers in animal production in the one-health context.

Stress and welfare are important factors in animal production in the context of growing production optimization and scrutiny by the general public. In a context in which animal and human health are intertwined aspects of the one-health concept it is of utmost importance to define the markers of stress and welfare. These are important tools for producers, retailers, regulatory agents and ultimately consumers to effectively monitor and assess the welfare state of production animals. Proteomics is the science that studies the proteins existing in a given tissue or fluid. In this review we address this topic by showing clear examples where proteomics has been used to study stress-induced changes at various levels. We adopt a multi-species (cattle, swine, small ruminants, poultry, fish and shellfish) approach under the effect of various stress inducers (handling, transport, management, nutritional, thermal and exposure to pollutants) clearly demonstrating how proteomics and systems biology are key elements to the study of stress and welfare in farm animals and powerful tools for animal welfare, health and productivity.

Animal board invited review: advances in proteomics for animal and food sciences.

Animal production and health (APH) is an important sector in the world economy, representing a large proportion of the budget of all member states in the European Union and in other continents. APH is a highly competitive sector with a strong emphasis on innovation and, albeit with country to country variations, on scientific research. Proteomics (the study of all proteins present in a given tissue or fluid - i.e. the proteome) has an enormous potential when applied to APH. Nevertheless, for a variety of reasons and in contrast to disciplines such as plant sciences or human biomedicine, such potential is only now being tapped. To counter such limited usage, 6 years ago we created a consortium dedicated to the applications of Proteomics to APH, specifically in the form of a Cooperation in Science and Technology (COST) Action, termed FA1002--Proteomics in Farm Animals: www.cost-faproteomics.org. In 4 years, the consortium quickly enlarged to a total of 31 countries in Europe, as well as Israel, Argentina, Australia and New Zealand. This article has a triple purpose. First, we aim to provide clear examples on the applications and benefits of the use of proteomics in all aspects related to APH. Second, we provide insights and possibilities on the new trends and objectives for APH proteomics applications and technologies for the years to come. Finally, we provide an overview and balance of the major activities and accomplishments of the COST Action on Farm Animal Proteomics. These include activities such as the organization of seminars, workshops and major scientific conferences, organization of summer schools, financing Short-Term Scientific Missions (STSMs) and the generation of scientific literature. Overall, the Action has attained all of the proposed objectives and has made considerable difference by putting proteomics on the global map for animal and veterinary researchers in general and by contributing significantly to reduce the East-West and North-South gaps existing in the European farm animal research. Future activities of significance in the field of scientific research, involving members of the action, as well as others, will likely be established in the future.

A discovery-phase urine proteomics investigation in type 1 diabetes.

Diabetes is a chronic metabolic disease which can lead to serious health problems particularly in and to the development of cardiovascular and renal complications. The aim of this study is to possibly identify distinctive molecular features in urine samples which might correlate to the progression and complications of type 1 diabetes. Diabetic patients with normo- and micro-albuminuria have been analyzed and compared to a group of control subjects. Urine proteins of control and type 1 diabetes subjects were investigated in their proteome profiles, using high-resolution two-dimensional gel electrophoresis separation and protein identifications by MALDI-TOF-MS and LC-MS/MS analysis. Proteomics analysis highlighted differential expression of several proteins between control and type 1 diabetes subjects. In particular, five proteins were found to be down-regulated and four proteins up-regulated. Lower protein representations in diabetic subjects were associated with Tamm-Horsfall urinary glycoprotein, apolipoprotein A-I, apolipoprotein E, α2-thiol proteinase inhibitor, and human complement regulatory protein CD59, while higher protein representations were found for α-1-microglobulin, zinc-α2 glycoprotein, α-1B glycoprotein, and retinol-binding protein 4. These differences were maintained comparing control subjects with type 1 diabetes normo-albuminuric and micro-albuminuric subjects. Furthermore, these proteins are correlated to glycosylated hemoglobin and microalbuminuria, confirming their role in diabetic pathology. This study gives new insights on potential molecular mechanisms associated with the complications of type 1 diabetic disease providing evidences of urine proteins potentially exploitable as putative prognostic biomarkers.

The use of fentanyl-patch in dogs undergoing spinal surgery: plasma concentration and analgesic efficacy.

Objectives of this study were to evaluate plasma concentrations and analgesic efficacy of fentanyl administered transdermically in dogs undergoing spinal surgery. At the end of the surgery and before awakening, a fentanyl-patch was applied and was maintained in situ for 72 h. Blood samples were taken before the application of the patch, at 2, 4, 6, 8, 10, 12, 18, 24, 32, 40, 48, 60, and 72 h after application and then 2, 4, 6, 8, 10, and 12 h after its removal. Before each blood sampling, pain evaluation was carried out using the Glasgow pain score, appropriately modified. Plasma concentrations of fentanyl were determined using a specific immuno-enzymatic kit. In this study, the minimum analgesic plasma concentration (0.23 ng/mL) required to achieve analgesia in human and considered to apply also for dogs was reached in all animals. No animal showed pain in the range of 'intense pain'; in two cases, the level of the pain was slight or moderate. No undesired effects were found. Results suggest that the use of transdermic patches could represent a valid aid in pain therapy in small animals; in particular, it contributes to the postoperative well-being of patients undergoing major surgery.

Proteomics to investigate fertility in bulls.

In dairy cattle breeding, herd reproductive management is the primary focus, affecting a large part of the general costs. A negative association was observed between the level of milk production and fertility. Some studies have shown that a significant percentage of reproductive failure is attributable to semen quality; therefore, if reproduction management is based on artificial insemination, then it is important to assess the fertility level of the sires. In this study, proteomic analysis was used to compare the protein expression profiles from sperm of high- and low-fertility bulls. Comparative proteomic analysis showed that expression of several proteins [nine different two-dimensional electrophoresis (2-DE) spots] is related to fertility level (p

Mannanoligosaccharides and aflatoxin B1 in feed for laying hens: effects on egg quality, aflatoxins B1 and M1 residues in eggs, and aflatoxin B1 levels in liver.

Ninety-six laying hens were allocated to 4 groups and fed diets (control diet (0-0), diet supplemented with 2.5 ppm aflatoxin B1 (0-AF); diet supplemented with 0.11% mannanoligosaccharide (MOS-0); diet supplemented with 0.11% MOS and 2.5 ppm aflatoxin B1 (MOS-AF) for 4 wk to evaluate the effect of aflatoxin B1 (AFB1), mannanoligosaccharide (MOS), or both on egg quality and the in vivo efficacy of MOS to interact with an oral administration of AFB1. After 2 and 3 wk, egg weight decreased (P < 0.05) in the group fed MOS-0 versus groups on 0-0 and 0-AF. Egg shell weight was lower (P < 0.05) in the group fed 0-AF. Aflatoxin influenced color parameters, which were probably related to interference of AFB1 with lipid metabolism and pigmentary substances deposition in yolk. MOS appeared to increase protein percentage in albumen. No AFB1 or aflatoxin M1 (AFM1; a polar metabolite of AFB1) residues were found in eggs of the experimental groups. Livers from groups 0-0 and MOS-0 always tested negative for AFB1 and AFM1. Differences (P < 0.01) were found between AFB1 hepatic levels of group 0-AF (mean +/- SD: 4.13 +/- 1.95 ppb) and group MOS-AF (mean +/- SD: 2.21 +/- 1.37 ppb). The data demonstrated the ability of MOS to adsorb and degrade AFB1, reducing gastrointestinal absorption of AFB1 and its levels in tissues.

Aflatoxin B1 and clinoptilolite in feed for laying hens: effects on egg quality, mycotoxin residues in livers, and hepatic mixed-function oxygenase activities.

Ninety-six laying hens were allocated to four groups administered different diets (group 0-0 received a complete diet, group 0-AF received a diet supplemented with 2.5 ppm of aflatoxin B1 [AFB1], group 2-0 received a diet supplemented with 2% clinoptilolite [CPL], and group 2-AF received a diet supplemented with 2% CPL and 2.5 ppm of AFB1) for 4 weeks to evaluate the effect of AFBI and/or CPL on egg quality and the ability of CPL to interact with the oral administration of AFB1. The possible effects of AFB1 on cytochrome P450-dependent hepatic mixed-function oxygenase (MFO) activities were also evaluated. Mycotoxin reduced yolk weight, while CPL influenced albumen percentage relative to that of eggs laid by chickens in group 0-AF Eggs laid by chickens in groups 0-AF and 2-AF had stronger shells and weighed less than the eggs of other groups. The eggs of treated groups were lighter in color than those of the control group (P < 0.01), and the tendency to yellowness in eggs was increased by CPL, probably through the affinity of red pigments for adsorbents and a consequent prevalence of yellow tonality. Color parameters might be connected with AFB1's interference with lipid metabolism and pigment deposition. The livers of hens in groups 0-AF and 2-AF showed very low mycotoxin concentrations that were significantly different (P < 0.01). The highest levels observed were those in the livers of the hens receiving the diet supplemented with the mycotoxin alone. AFB1 did not exert any significant effects on the hepatic MFO activities examined.

Pharmacokinetics and residual behaviour in milk of oxytetracycline in cows following administration of uterine pessaries.

The plasma kinetics and residual depletion in milk of cows treated by the intrauterine route with pessaries containing oxytetracycline (OTC) were evaluated. The antibiotic was administered to five healthy Friesian cows at a dosage of 3g/head in the early post partum phase. Blood samples were collected before and at different time intervals (3, 6, 12, 24, 48, 72, 84, and 96 h) after treatment. Milk was drawn before treatment and at 12-h intervals for 4 consecutive days. Samples were analysed by a high-performance liquid chromatography method and the pharmacokinetic parameters were processed using the minimum Akaike information criterion estimation (MAICE) test. The mean values obtained indicated a relatively low area under the concentration time curve (25.19+/-12.61 microg/mg per h) and maximum plasma concentration (Cmax) (0.549+/-0.278 microg/mL) with delayed time to Cmax (11.71+/-4.15 h) and elimination half-life (21.96+/-4.42 h). A similar pattern could be shown for milk, in which measurable residual levels are found in two out of five animals until the 72nd hour after treatment. Data obtained demonstrate that OTC administered as a solid form is poorly and slowly absorbed from the uterus of cows.

Milk depletion of dicloxacillin residues in cows and sheep following intramammary administration.

The excretion rate of dicloxacillin from milk was studied after intramammary administration of a suspension of the drug active in vegetable oil. Eight cows and eight sheep, four of each group in low and four in high milk production, were dosed with 200 mg dicloxacillin/quarter in cows and 100 mg dicloxacillin/quarter in sheep, three times at 12 h intervals. The dicloxacillin concentrations in milk were quantified by high performance liquid chromatography (HPLC). In cows, time until dicloxacillin was undetectable was 48 h and no difference was observed between the groups. In sheep, dicloxacillin was undetectable 72 h and 84 h after the treatment in low and in high milk production groups, respectively. The implications of several factors affecting the possible milk withdrawal period were studied. The results indicated that the pharmaceutical vehicle and the coefficient of lipid solubility exerted major effects on depletion time.