Detection of pathogens from environmental samples for the evaluation of the health status of laboratory animals

Pathogens present in the environment are the biggest source of diseases and epidemics in the breeding of laboratory animals. The presence of microorganisms, in fact, can critically influence the animal health status and consequently the validity and reproducibility of experimental data. In accordance with the 3Rs principle, this study fits into the Refinement and Reduction concepts. The development of a health surveillance plan on environmental material from animal housing would have an important impact not only on maintaining an adequate state of health and on the generation of quality experimental data, but also on reducing the number of animals to be sacrificed.

Large Farm Animals Used for Research Purposes: A Survey on Purchase, Housing and Hygiene Management

Background: Farm animals (FAs) are frequently used in biomedical research. Recommendations for the purchase, housing and health monitoring of these animals (sheep, goats, cattle and pigs) are still missing, and many institutes have developed their own strategies and protocols to face the challenges associated with the use of farm animals. This may influence the comparability of research results and increase data variances, thus increasing animal use that contradicts the obligation to apply the 3Rs principle of reduction, refinement and replacement required in Directive 2010/63 EU and the German animal protection law. Methods: A survey was conducted to define the current state of the art in research institutes working with pigs, and large and small ruminants. Results: The results of the survey clearly show that there are no uniform procedures regarding the purchase, housing and hygiene management of farm animals contrary to small laboratory animals. The facilities make purpose-bound decisions according to their own needs and individual work instructions and implement their own useful protocols to improve and maintain the health of the animals. Conclusion: This survey was the first step to filling the gaps and identifying the status quo and practical applied measures regarding the purchase and hygiene monitoring of FAs in order to improve animal welfare and scientific validity.

The 3Rs Principle in Animal Experimentation: A Legal Review of the State of the Art in Europe and the Case in Italy

The aim of this paper is to describe the essential points of Italian and European legislation governing the use of animals in biomedical experimentation. A close look will be taken at the principles of the 3Rs, which represent the mainstay of the legal architecture based on which a correct interpretation may be drawn of the legislative documents on animal experimentation. Furthermore, this paper will address the ways in which Directive 2010/63/EU is implemented in Italian legislation on the welfare of laboratory animals. In addition to an assessment of legal issues (such as the scope of jurisdiction of supervisory authorities tasked with issuing authorizations), it will include a discussion of cases of inadequate and insufficient implementation of the requirements laid down by Directive 2010/63/EU. Both the consistency of the interpretation of national legislation with the Directive and the direct effectiveness of the Directive in national law, in which animal testing has been and still is the subject of heated debate between supporters and opponents, will be examined.

In Vitro Tests for Assessing the Neutralizing Ability of Snake Antivenoms: Toward the 3Rs Principles

There is an urgent need to strengthen the implementation of the 3Rs principle (Replacement, Reduction and Refinement) in the use of experimental animals in toxinological research and in the assessment of the neutralizing efficacy of snake antivenoms. This is a challenging task owing to the inherent complexity of snake venoms. The state of the art on this topic is hereby reviewed, with emphasis on the studies in which a correlation has been observed between in vivo toxicity tests and in vitro surrogate assays, particularly in the study of lethal activity of venoms and its neutralization. Correlations have been described with some venoms-antivenoms when using: (a) enzyme immunoassays, (b) hemagglutination, (c) enzyme assays (proteinase, phospholipase A2), (d) in vitro coagulant effect on plasma, (e) cell culture assays for cytotoxicity, (f) functional assays for assessing neurotoxicity in vitro, (g) use of hens’ eggs, and (h) antivenomics. Additionally, the routine introduction of analgesia in these assays and the design of more ‘humane’ protocols for the lethality test are being pursued. It is expected that the next years will witness a growing awareness of the relevance of the 3Rs principles in antivenom testing, and that new in vitro alternatives and more ‘humane’ experimental designs will emerge in this field.

Health monitoring program for the control of laboratory animal diseases

Pathogens present in the environment are the biggest source of diseases and epidemics in the breeding of laboratory animals. In fact, the presence of microorganisms can critically influence the animal health status and, consequently, the validity and reproducibility of experimental data. In accordance with the 3Rs principle (Refinement, Reduction, Replacement), this study is part of the Refinement concept. The FELASA guidelines, formulated with the aim of guaranteeing the best animal health state, are a valid support for researchers. In this preliminary study, health-monitoring program was carried out within the breeding of laboratory animals in IZSLER facility. The main murine viruses were analyzed through molecular biology techniques (PCR, RT-PCR) and enzyme immunoassays (indirect ELISA). The established surveillance program steadily guarantees animal health and ensures the most controlled environmental and sanitary conditions. Further investigations will be needed to develop virus control strategies.

Zebrafish as an alternative method for toxicological studies

According to the Directive 2010/63/EU fish embryos do not fall into regulatory frameworks dealing with animal experimentation. Therefore, in compliance with the 3Rs principle, zebrafish embryos are considered as replacement or refinement methods. Since more and more industrial chemicals are recognized causes of skin sensitization, it is needed a thorough understanding of the mechanisms to make predictions of the toxic potential of novel compounds. Thus, the FET test was performed and up to four apical observations were recorded as indicators of lethality: coagulation of fertilized eggs, lack of somite formation, no detachment of the tail bud from the yolk sac and lack of heartbeat. Then, in order to assess whether the skin sensitization due to chemical incubation was really measurable, the Fish Interleukin 8 (IL8) ELISA Kit was carried out. The preliminary results obtained so far seem encouraging. However, they need to be confirmed through further ELISA tests and compared with other in vitro methods.

GAMYBOS LOGISTIKOS TOBULINIMAS BIOEKONOMIKOS IŠŠŪKIŲ KONTEKSTE / IMPROVEMENT OF PRODUCTION LOGISTICS IN THE CONTEXT OF BIOECONOMIC CHALLENGES

In the context of globalization, with a rapid drop in natural resources and an increase in the amount of waste an effective tool for boosting business competitiveness is the bioeconomy. The purpose of the article is to review the standpoint of scientists and different institutions towards the concept of bioeconomy and evaluate the use of circular economy in industry. The study proposed a production logistics model that is based on the use of the circular economy and the 3Rs principle. Based on the results of the research, the potential benefits of using the circular economy have been identified, which will encourage resource conservation and waste reduction. Santrauka Globalizacijos kontekste sparčiai senkant gamtiniams ištekliams ir didėjant atliekų kiekiui, efektyvi priemonė verslo konkurencingumui didinti yra bioekonomika. Straipsnio tikslas – apžvelgti mokslininkų ir skirtingų institucijų požiūrį į bioekonomikos sampratą ir įvertinti žiedinės ekonomikos naudojimą pramonėje. Tyrimo metu buvo pasiūlytas gamybos logistikos modelis, grindžiamas žiedinės ekonomikos ir 3Rs principo taikymu. Remiantis tyrimų rezultatais buvo nustatyti žiedinės ekonomikos naudojimo pranašumai, kurie skatins tausoti išteklius ir mažinti atliekas.