Development of a Training Strategy Aimed at Increasing Veterinarians' Awareness of the Proper Use of Antibiotics on Rabbit Farms.

The rabbit sector faces significant challenges with antimicrobial usage (AMU) and antimicrobial resistance (AMR). A focus group involving nine rabbit sector stakeholders identified key issues contributing to high AMU and the need for veterinarians' training. Participants emphasized the lack of clear legislation, biosecurity standards, and the importance of training on best practices to reduce AMU. To address these concerns, a blended learning course for rabbit veterinarians was organized, focusing on responsible drug use to lower AMU. The course design promoted participant interaction using group exercises and online discussions. The output was an operational tool, encompassing crucial elements to reduce drug dependence, covering housing, environmental conditions, feed, reproduction, disease prevention, diagnosis, and treatments. Validated in veterinarians' daily practices, the tool proved valuable. The tool, presented as a checklist, assists veterinarians in supporting field activities regarding proper drug use and AMR issues. It also helps farmers address knowledge gaps in breeding management and adopt biosecurity practices for disease prevention. Participants were encouraged to use the tool during farm visits and collaborate with farmers. The project presented in the manuscript is a crucial step towards the development of effective strategies for responsible AMU and AMR mitigation in the rabbit sector and beyond.

Data distribution in public veterinary service: health and safety challenges push for context-aware systems.

BACKGROUND: Today's globalised and interconnected world is characterized by intertwined and quickly evolving relationships between animals, humans and their environment and by an escalating number of accessible data for public health. The public veterinary services must exploit new modeling and decision strategies to face these changes. The organization and control of data flows have become crucial to effectively evaluate the evolution and safety concerns of a given situation in the territory. This paper discusses what is needed to develop modern strategies to optimize data distribution to the stakeholders. MAIN TEXT: If traditionally the system manager and knowledge engineer have been concerned with the increase of speed of data flow and the improvement of data quality, nowadays they need to worry about data overflow as well. To avoid this risk an information system should be capable of selecting the data which need to be shown to the human operator. In this perspective, two aspects need to be distinguished: data classification vs data distribution. Data classification is the problem of organizing data depending on what they refer to and on the way they are obtained; data distribution is the problem of selecting which data is accessible to which stakeholder. Data classification can be established and implemented via ontological analysis and formal logic but we claim that a context-based selection of data should be integrated in the data distribution application. Data distribution should provide these new features: (a) the organization of situation types distinguishing at least ordinary vs extraordinary scenarios (contextualization of scenarios); (b) the possibility to focus on the data that are really important in a given scenario (data contextualization by scenarios); and (c) the classification of which data is relevant to which stakeholder (data contextualization by users). SHORT CONCLUSION: Public veterinary services, to efficaciously and efficiently manage the information needed for today's health and safety challenges, should contextualize and filter the continuous and growing flow of data by setting suitable frameworks to classify data, users' roles and possible situations.

Information and organization in public health institutes: an ontology-based modeling of the entities in the reception-analysis-report phases.

BACKGROUND: Ontologies are widely used both in the life sciences and in the management of public and private companies. Typically, the different offices in an organization develop their own models and related ontologies to capture specific tasks and goals. Although there might be an overall coordination, the use of distinct ontologies can jeopardize the integration of data across the organization since data sharing and reusability are sensitive to modeling choices. RESULTS: The paper provides a study of the entities that are typically found at the reception, analysis and report phases in public institutes in the life science domain. Ontological considerations and techniques are introduced and their implementation exemplified by studying the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), a public veterinarian institute with different geographical locations and several laboratories. Different modeling issues are discussed like the identification and characterization of the main entities in these phases; the classification of the (types of) data; the clarification of the contexts and the roles of the involved entities. The study is based on a foundational ontology and shows how it can be extended to a comprehensive and coherent framework comprising the different institute's roles, processes and data. In particular, it shows how to use notions lying at the borderline between ontology and applications, like that of knowledge object. The paper aims to help the modeler to understand the core viewpoint of the organization and to improve data transparency. CONCLUSIONS: The study shows that the entities at play can be analyzed within a single ontological perspective allowing us to isolate a single ontological framework for the whole organization. This facilitates the development of coherent representations of the entities and related data, and fosters the use of integrated software for data management and reasoning across the company.

Confirmation of protein biomarkers of corticosteroids treatment in veal calves sampled under field conditions.

In veal calf production, growth promoters are still illicitly used. Surveillance of misuse of such molecules is necessary to preserve human health. Methods currently adopted for their analysis are based on liquid chromatography-tandem mass spectrometry, but their efficacy can be affected by undetectable residual concentrations in biological matrices due to treatments at low-dosage or based on unknown anabolic compounds. The development of screening methods to identify the indirect biological effects of administration of growth promoters can improve the efficiency of drug residue monitoring. To this purpose, an integrated approach has been used to further validate the set of protein biomarkers defined in a previous controlled study to detect the use of corticosteroids through the changes caused in muscle protein expression. The thymus morphology of 48 samples collected under field conditions was evaluated to assess the presence of potential corticosteroids treatment. Animals were divided on the basis of their thymus characteristics in negative or suspected for illegal corticosteroids treatment. Drug residue analyses were performed on the liver, giving a satisfactory correlation with the histological examination (∼85%). Finally, the proteomics analysis of muscle protein extracts was carried out by 2D differential in gel electrophoresis, and proteins that were differentially expressed between the two animal groups (p value <0.01) were selected for MALDI-MS/MS analysis. This approach allowed us to identify 29 different proteins, and our findings indicate that the altered protein expression pattern can be used as an indirect method for the detection of illicit corticosteroids administration. A subset of the identified proteins was already reported in a previous controlled study, proving that these biomarkers can be used to develop a screening assay to improve the tools currently available for the detection of corticosteroids abuse in bovine meat production.

Sub-therapeutic treatments of bulls with dexamethasone: direct and indirect markers of treatment.

Dexamethasone is a potent synthetic corticosteroid synthesised as an anti-inflammatory agent, but also illegally used at low dosages as a growth promoter either alone or in combination with other steroids or ß-agonists. The use of screening methods based on the detection of biological effects of growth promoters is a promising approach to assist residue monitoring. The advantage of such an approach is that it offers the possibility to discover illicit treatments even when the drug residues are not easily detectable by common analytical methods. The aim of this study was to determine the urinary elimination and tissue distribution of dexamethasone and to investigate whether endogenous corticosteroid profiling of urine could be used to establish a predictive tool for identifying any misuse of growth promoters in bulls. Urinary corticosteroid concentrations were assayed in bulls (n = 8) treated with dexamethasone (0.75 mg/animal/day), bulls (n = 8) treated with a combination of dexamethasone (0.66 mg/animal/day) and clenbuterol (from 2 to 6 mg/animal/day) and untreated bulls (n = 8). The distribution and elimination of dexamethasone and the urinary concentration of cortisone, cortisol and 6ß-hydroxycortisol were determined by using LC-MS/MS at different time points both during treatments and after appropriate withdrawal times. The urinary concentration of endogenous corticosteroids in bulls decreased significantly (p < 0.001) upon the administration of dexamethasone alone, whereas it remained unchanged until the last sampling point in animals treated with dexamethasone and clenbuterol (p < 0.001). Nonetheless, dexamethasone administration significantly reduced (p < 0.01) the urinary 6ß-hydroxycortisol/cortisol ratio in both treatment groups. Therefore, this ratio could be regarded as a candidate for an indirect screening assay for detecting glucocorticoid abuse in bull fattening to pick out farms where similar treatments are potentially in use and to suggest targeted sampling on remaining animals.

Evaluation of thymus morphology and serum cortisol concentration as indirect biomarkers to detect low-dose dexamethasone illegal treatment in beef cattle.

BACKGROUND: Corticosteroids are illegally used in several countries as growth promoters in veal calves and beef cattle, either alone or in association with sex steroids and ß-agonists, especially at low dosages and primarily through oral administration, in order to enhance carcasses and meat quality traits. The aim of the present study is to evaluate the reliability of the histological evaluation of the thymus, as well as the serum cortisol determination, in identifying beef cattle, treated with two different dexamethasone-based growth-promoting protocols and the application of different withdrawal times before slaughter. RESULTS: Our findings demonstrate that low dosages of dexamethasone (DXM), administered alone or in association with clenbuterol as growth promoter in beef cattle, induce morphologic changes in the thymus, resulting in increase fat infiltration with concurrent cortical atrophy and reduction of the cortex/medulla ratio (C/M). In fact, the C/M value was significantly lower in treated animals than in control ones, with both the protocols applied. The cut off value of 0.93 for the cortex/medulla ratio resulted to be highly effective to distinguish control and treated animals. The animals treated with DXM showed inhibition of cortisol secretion during the treatment period, as well as at the slaughterhouse, 3 days after treatment suspension. The animals treated with lower doses of DXM in association with clenbuterol, showed inhibition of cortisol secretion during the treatment period, but serum cortisol concentration was restored to physiological levels at slaughterhouse, 8 days after treatment suspension. CONCLUSIONS: The histological evaluation of thymus morphology, and particularly of the C/M may represent a valuable and reproducible method applicable to large-scale screening programs, due to the easy sampling procedures at slaughterhouse, as well as time and cost-saving of the analysis. Serum cortisol determination could be considered as an useful in vivo biomarker of dexamethasone illegal treatment in beef cattle during the fattening period, whilst it does not appear to be a good biomarker at the slaughterhouse, since the protocol of DXM administration, as well as the withdrawal period could affect the reliability of the method.

Protein expression changes in skeletal muscle in response to growth promoter abuse in beef cattle.

The fraudulent treatment of cattle with growth promoting agents (GPAs) is a matter of great concern for the European Union (EU) authorities and consumers. It has been estimated that 10% of animals are being illegally treated in the EU. In contrast, only a much lower percentage of animals (<0.5%) are actually found as being noncompliant by conventional analytical methods. Thus, it has been proposed that methods should be developed that can detect the use of the substances via the biological effects of these substances on target organs, such as the alteration of protein expression profiles. Here we present a study aimed at evaluating if a correlation exists between the treatment with GPAs and alterations in the two-dimensional electrophoresis (2DE) protein pattern obtained from the biceps brachii skeletal muscle from mixed-bred cattle. After image analysis and statistical evaluation, protein spots that differentiate between treated and control groups were selected for analysis by mass spectrometry. A set of proteins could be defined that accurately detect the use of glucocorticoids and ß(2)-agonists as growth promoters through the changes caused in muscle differentiation. As a further validation, we repeated the analysis using an independent set of samples from a strain of pure-bred cattle and verified these proteins by Western blot analysis.

Steroidogenic enzyme gene expression profiles in the testis of cattle treated with illicit growth promoters.

Recently, the effect of illicit growth promoters (GPs) upon the cattle transcriptome has drawn the increasing attention of the scientific community. In the present study, the pre-transcriptional effects of three different illicit protocols on a set of target genes, including steroidogenic enzymes and three related transcription factors, were estimated in cattle testis. Beef cattle were administered with dexamethasone (DEX) orally (group D(1)) or intramuscularly in experiment 1 (group DIM). In experiment 2, DEX was orally administered alone (group D(2)) or with 17ß-estradiol (group DE), and in experiment 3, dehydroepiandrosterone and boldione were orally administered alone (group DHEA and group ADD) or in combination (group DHAD). The GP effects were measured by quantitative real time RT-PCR. The results of our study were significant but not univocal. A GP-dependent effect on target gene mRNA levels was noticed for 3ß-hydroxysteroid dehydrogenase type 1 (HSD3ß1,p<0.05 and p<0.01 for the D(2), DE and DHAD groups, respectively), the cytochrome P450 side chain cleavage (DHAD, p<0.05), the cytochrome P450 17A1 (DIM and D(2), p<0.05), HSD17ß3 (DE, p<0.05), aromatase (DHEA, p<0.05), the androgen receptor (DHAD, p<0.05) and the mineralocorticoid receptor-like (DIM, p<0.05). Our present results suggest that different GP schedules are likely to affect genes involved in steroid synthesis and regulation in cattle testis. Thus, this tissue might be considered a potential surrogate tissue that warrants further study into its usefulness in the screening of GP abuse.