Non-Technical Project Summaries (NTS) are legal documents that were first introduced by the to enhance transparency within scientific animal experimentation. Researchers intending to conduct biological research on anima...Non-Technical Project Summaries (NTS) are legal documents that were first introduced by the to enhance transparency within scientific animal experimentation. Researchers intending to conduct biological research on animal models must fulfil the NTS requirements by outlining their proposed use of animals and how they plan to implement the Three Rs (replacement, reduction and refinement of animal use) in their experiments. This study outlines a novel systematic analysis approach that enables the assessment of NTS transparency based on the accuracy of reporting of certain Three Rs-specific information. This potentially customisable strategy could help toward the development of practical guidelines for use by Animal Welfare and Ethical Review Bodies (AWERBs) in establishments conducting animal research, in the process of scrutinising NTS during their pre-submission review of proposed licence applications. This could help to identify gaps in reporting of Three Rs-specific information relating to the planned animal experiments, which represents a remarkable step toward achieving greater openness in scientific communication. This study supports the concept that NTS transparency can promote the implementation of non-animal alternatives in fields where this is currently lacking, such as respiratory disease research. Although NTS were originally conceived as informative documents for a lay audience, we can conclude that data in NTS can be successfully used as a basis for systematic analysis. By reviewing the NTS, the experimental limitations of the currently available replacement strategies can also be highlighted, potentially pinpointing where there is a need for future method development.
Vero cells are one of the most frequently used cell types in virology. They can be used not only as a vehicle for the replication of viruses, but also as a model for investigating viral infectivity, cytopathology and vac...Vero cells are one of the most frequently used cell types in virology. They can be used not only as a vehicle for the replication of viruses, but also as a model for investigating viral infectivity, cytopathology and vaccine production. There is increasing awareness of the need to limit the use of animal-derived components in cell culture media for a number of reasons, which include reducing the risk of contamination and decreasing costs related to the downstream processing of commercial products obtained via cell culture. The current study evaluates the use of protein hydrolysates (PHLs), also known as peptones, as partial substitutes for fetal bovine serum (FBS) in Vero cell culture. Eleven plant-based, two yeast-based, and three casein-based peptones were assessed, with different batches evaluated in the study. We tested the effects of three concentration ratios of FBS and peptone on Vero cell proliferation, four days after the initial cell seeding. Some of the tested peptones, when in combination with a minimal 1% level of FBS, supported cell proliferation rates equivalent to those achieved with 10% FBS. Collectively, our findings showed that plant-based peptones could represent promising options for the successful formulation of serum-reduced cell culture media for vaccine production. This is especially relevant in the context of the current COVID-19 pandemic, in view of the urgent need for SARS-CoV-2 virus production for certain types of vaccine. The current study contributes to the Three Rs principle of , as well as addressing animal ethics concerns associated with FBS, by repurposing PHLs for use in cell culture.
The contemporary pharmaceutical industry is voicing growing concerns about the translatability and reproducibility of animal models. In addition, the usefulness of certain of the required regulatory safety tests in anima...The contemporary pharmaceutical industry is voicing growing concerns about the translatability and reproducibility of animal models. In addition, the usefulness of certain of the required regulatory safety tests in animals is being increasingly questioned. It remains difficult, however, to make the move toward alternative testing methods, not least because of legislative demands. A historical analysis was performed, in order to study how the mandatory animal studies in legislative requirements came about. This article reflects on the role that specific public health disasters played in the creation of (more) regulatory requirements for animal testing. It will show how the regulatory changes prompted by the sulfanilamide elixir disaster in the 1930s and the thalidomide disaster in the early 1960s were based on the belief that extensive animal testing would prevent similar future human health tragedies. As scientists increasingly highlight issues with translatability between non-human animals and humans, the belief that current regulatory requirements ensure safety becomes more difficult to maintain. In addition, it means that some of the regulations now in place require animal tests that do not contribute to the safety of a drug, as shown in a third case study of the court case by Vanda industries against the FDA. We finally argue that regulations should be critically examined and altered where necessary, so that they are no longer a barrier in the transition toward animal-free testing and more human-relevant science.
Cell culture techniques are strongly connected with modern scientific laboratories and production facilities. Thus, choosing the most suitable medium for the cells involved is vital, not only directly to optimise cell vi...Cell culture techniques are strongly connected with modern scientific laboratories and production facilities. Thus, choosing the most suitable medium for the cells involved is vital, not only directly to optimise cell viability but also indirectly to maximise the reliability of the experiments performed with the cells. Fetal bovine or calf serum (FBS or FCS, respectively) is the most commonly used cell culture medium supplement, providing various nutritional factors and macromolecules essential for cell growth. Yet, the use of FBS encompasses a number of disadvantages. Scientifically, one of the most severe disadvantages is the lot-to-lot variability of animal sera that hampers reproducibility. Therefore, transitioning from the use of these ill-defined, component-variable, inconsistent, xenogenic, ethically questionable and even potentially infectious media supplements, is key to achieving better data reproducibility and thus better science. To demonstrate that the transition to animal component-free cell culture is possible and achievable, we highlight three different scenarios and provide some case studies of each, namely: i) the adaptation of single cell lines to animal component-free culture conditions by the replacement of FBS and trypsin; ii) the adaptation of multicellular models to FBS-free conditions; and (iii) the replacement of FBS with human platelet lysate (hPL) for the generation of primary stem/stromal cell cultures for clinical purposes. By highlighting these examples, we aim to foster and support the global movement towards more consistent science and provide evidence that it is indeed possible to step out of the currently smouldering scientific reproducibility crisis.
There is ethical debate over whether fetal calves suffer when their dam is slaughtered and fetal blood extracted by cardiac puncture for fetal bovine serum (FBS) production. Yet, the serum industry does not follow best p...There is ethical debate over whether fetal calves suffer when their dam is slaughtered and fetal blood extracted by cardiac puncture for fetal bovine serum (FBS) production. Yet, the serum industry does not follow best practice, as recommended by the European Food Safety Authority (EFSA), to avoid fetal distress. We discuss the key elements of this debate, and recommend how the serum industry can alter its practices to improve animal welfare.
Behrsing HP, Wahab A, Ukishima L
… +15 more, Grodi C, Frentzel S, Johne S, Ishikawa S, Ito S, Wieczorek R, Budde J, Keyser BM, Haswell LE, Thorne D, Bishop E, Breheny D, Cao X, Qin Q, Patel V
The use of reconstituted human airway (RHuA) epithelial tissues to assess functional endpoints is highly relevant in respiratory toxicology, but standardised methods are lacking. In June 2015, the Institute for In Vitro...The use of reconstituted human airway (RHuA) epithelial tissues to assess functional endpoints is highly relevant in respiratory toxicology, but standardised methods are lacking. In June 2015, the Institute for In Vitro Sciences (IIVS) held a technical workshop to evaluate the potential for standardisation of methods, including ciliary beat frequency (CBF). The applicability of a protocol suggested in the workshop was assessed in a multi-laboratory ring study. This report summarises the findings, and uses the similarities and differences identified between the laboratories to make recommendations for researchers in the absence of a validated method. Two software platforms for the assessment of CBF were used - Sisson-Ammons Video Analysis (SAVA; Ammons Engineering, Clio, MI, USA) and ciliaFA (National Institutes of Health, Bethesda, MD, USA). Both were utilised for multiple read temperatures, one objective strength (10×) and up to four video captures per tissue, to assess their utility. Two commercial RHuA tissue cultures were used: MucilAir™ (Epithelix, Geneva, Switzerland) and EpiAirway™ (MatTek, Ashland, MA, USA). IL-13 and procaterol were used to induce CBF-specific responses as positive controls. Further testing addressed the impact of tissue acclimation duration, the number of capture fields and objective strengths on baseline CBF readings. Both SAVA and ciliaFA reliably collected CBF data. However, ciliaFA failed to generate accurate CBF measurements above ∼10 Hz. The positive controls were effective, but were subject to inter-laboratory variability. CBF endpoints were generally uniform across replicate tissues, objective strengths and laboratories. Longer tissue acclimation increased the percentage active area, but had minimal impact on CBF. Taken together, these findings support the development and validation of a standardised CBF measurement protocol.
Resources from biobanks and biorepositories, such as human samples, are of increasing interest to specialists in various fields. However, whilst biobanks provide a crucial service, their efficient and effective managemen...Resources from biobanks and biorepositories, such as human samples, are of increasing interest to specialists in various fields. However, whilst biobanks provide a crucial service, their efficient and effective management can prove challenging. When establishing a biobank many factors should be considered, such as the need for appropriate infrastructure, equipment, financial support, and highly specialised and suitably qualified personnel. The number and qualifications of the necessary personnel depend both on the biobank's size and type - i.e. a biobank that is large and diversified in terms of the stored material should be organised differently to a small biorepository. The core of the biobank should be composed of highly trained personnel that closely co-operate with the general and quality control manager. Due to the large amount of data related to the samples, an IT specialist might be needed. In the case of large population biobanks, personnel responsible for patient recruitment, documentation handling, sample collection and distribution to the biobank would be necessary. Furthermore, staff responsible for the infrastructure are also highly important, as they are the first responders to failures that may be critical for the biobank functioning. Depending on the type and size of the biobank/biorepository, some responsibilities and tasks could potentially be combined. Nevertheless, highly trained personnel with clear and precisely defined duties are the key to the proper functioning of a biobank.
Altern Lab Anim
· 2022 Jul · PMID 35801971
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Full text
While laboratory animals are necessary for some aspects of the development of scientific and biomedical advances, including those of precision medicine, the use of human tissues is necessary in order to explore the findi...While laboratory animals are necessary for some aspects of the development of scientific and biomedical advances, including those of precision medicine, the use of human tissues is necessary in order to explore the findings and ensure that they are relevant to human systems. Many sources of human tissues exist, but researchers - particularly those making the transition from animal to human systems - may not be aware of how best to find quality sources of human tissues or how best to use them in their research. In this article, we discuss the advantages of using human tissues in research. In addition, we highlight some of the major advances made possible through the use of human tissue, and describe how human tissue is collected for research. We discuss the various types of bioresources that make human tissue available, and advise on how investigators can find and use appropriate bioresources to support their research - with the hope that this information will help facilitate the transition from research on animals to research using human tissues, as rapidly as is practicable.
Colorectal cancer (CRC) is a global cause of cancer-related mortality driven by genetic and environmental factors which influence therapeutic outcomes. The emergence of next-generation sequencing technologies enables the...Colorectal cancer (CRC) is a global cause of cancer-related mortality driven by genetic and environmental factors which influence therapeutic outcomes. The emergence of next-generation sequencing technologies enables the rapid and extensive collection and curation of genetic data for each cancer type into clinical gene expression biobanks. We report the application of bioinformatics tools for investigating the expression patterns and prognostic significance of three genes that are commonly dysregulated in colon cancer: adenomatous polyposis coli (); B-Raf proto-oncogene (); and Kirsten rat sarcoma viral oncogene homologue (). Through the use of bioinformatics tools, we show the patterns of , and genetic alterations and their role in patient prognosis. Our results show mutation types, the frequency of mutations, tumour anatomical location and differential expression patterns for and for colorectal tumour and matched healthy tissue. The prognostic value of , and genetic alterations was investigated as a function of their expression levels in CRC. In the era of precision medicine, with significant advancements in biobanking and data curation, there is significant scope to use existing clinical data sets for evaluating the role of mutational drivers in carcinogenesis. This approach offers the potential for studying combinations of less well-known genes and the discovery of novel biomarkers, or for studying the association between various effector proteins and pathways.
There is a need for paradigm change in the methodology employed for toxicological testing and assessment. It could be said that this change is well on its way, through an evolutionary progress analogous to that of natura...There is a need for paradigm change in the methodology employed for toxicological testing and assessment. It could be said that this change is well on its way, through an evolutionary progress analogous to that of natural selection. Darwin's Theory of Evolution has defined the idea of evolution and descendancy since the last third of the 19th century. Increasingly, this concept of 'evolution' is being applied beyond the field of biology. This article discusses the progress of toxicological testing in the context of 'evolutionary pressure' and deliberates how this process can help foster the development, implementation and acceptance of mechanistic and human-relevant methods in this field. By comparing the current regulatory landscape in toxicity testing and assessment to specific elements in Charles Darwin's evolutionary theory, we aim to better understand the needs and requirements for the future.
The use of animals in research and education is a controversial topic that has raised extensive debates. Undergraduate students ( = 404) and lecturers ( = 62) from biomedical science schools at the Federal University of...The use of animals in research and education is a controversial topic that has raised extensive debates. Undergraduate students ( = 404) and lecturers ( = 62) from biomedical science schools at the Federal University of Goiás (UFG) in the municipality of Goiânia, Jataí and Catalão, Goiás, Brazil, were asked about their knowledge and opinions on bioethics, the use and importance of animals in education, the replacement of animal use with non-animal alternatives, and the current legislation of the National Council for the Control of Animal Experimentation (CONCEA) that bans animal use in some practical classes within technical and higher education (i.e. ). Most students and lecturers agreed not only that animal use can contribute to education, but also that it is important to replace this animal use with innovative non-animal alternatives where appropriate. The lecturers emphasised that the replacement of animal models will be possible only with the provision of appropriate training to improve the skills of educators in their use, as well as ensuring reliable access to suitable facilities and materials.
Animal dissection remains a common practice in American biology classrooms, despite the availability of dissection alternatives to study anatomy and physiology. Indeed, there is a growing body of evidence in the literatu...Animal dissection remains a common practice in American biology classrooms, despite the availability of dissection alternatives to study anatomy and physiology. Indeed, there is a growing body of evidence in the literature suggesting that the use of alternatives leads to the greater achievement of learning objectives, as compared to dissection. To better understand the current use of and attitudes toward dissection and alternatives, a nationwide survey of (mainly high-school) biology teachers ( = 2687) was conducted. Most educators believed that learning objectives related to biology subject content could be met through the use of alternatives, yet they preferred the hands-on experience of dissection. Most educators allow their students to use alternatives if requested, although few teachers ask students about their preference for using an animal specimen an alternative. Educators cited student engagement as the main factor driving their decision to choose between dissection specimens and alternatives, and felt that cost is the biggest barrier to implementing alternatives at their schools. Additional perspectives on dissection and alternatives were shared by survey participants. Since alternatives can be used to meet learning objectives associated with dissection, we recommend their use as replacements for traditional animal specimens, in line with the , and of animal use in education, according to the Three Rs principles.
In the context of veterinary education, simulators are devices or sets of conditions aiming to imitate real patients and enable students to practice skills without the need for live animal use. Simulator use in veterinar...In the context of veterinary education, simulators are devices or sets of conditions aiming to imitate real patients and enable students to practice skills without the need for live animal use. Simulator use in veterinary education has increased significantly in recent years, allowing consistent practical teaching without reliance on clinical cases. This review examines the available literature regarding the use of simulation and simulators for teaching practical day one competences to veterinary students. Scientific databases were searched and 73 relevant articles were reviewed. The reviewed articles revealed that there are a number of simulators currently available to veterinary educators, that simulators can enhance student skills and provide an alternative learning environment without the need for live animal and/or cadaver use, and that they usually receive positive feedback from the students who use them. There appears to be a bias towards small animal simulators - however, some skills that are developed through the use of small animal or table-top models will be transferrable to other species. The majority of large animal simulators focus on bovine rectal palpation and/or pregnancy diagnosis. Further research is required to increase the repertoire of available simulators for use in veterinary education, in order to improve the practical skills of veterinary students and reduce the use of live animals and cadaver material for teaching purposes.
The acquisition of ultrasound diagnostic skills via training is important for undergraduate veterinary medical students. Unfortunately, commercial simulators are costly, which limits their use and makes it necessary to d...The acquisition of ultrasound diagnostic skills via training is important for undergraduate veterinary medical students. Unfortunately, commercial simulators are costly, which limits their use and makes it necessary to develop low-cost simulators for training purposes. The aim of this study was to evaluate the efficacy of an easily constructed, low-cost, high-fidelity renal simulator for use in diagnostic ultrasound training of veterinary medical students. To construct the simulators, donated cat kidneys were embedded in a prepared agar matrix. The echogenicity of ultrasound scans obtained on the kidney simulator was assessed by the subject lecturers and compared with images acquired during clinical routine diagnostic procedures. Five students with no prior experience of the technique, under the direct supervision of a lecturer, performed five B-mode ultrasound examinations of the renal structure of the simulator. The structure of the kidney was assessed, and its length, width and thickness were measured. Three lecturers performed the same procedure as the students, and their results were used for comparison. Appropriate anatomical and ultrasonographic realism was achieved for each of the three layers of the kidney (cortex, medulla and pelvis), and similar pixel values were obtained with the simulator model and actual clinical diagnostic ultrasounds. In addition, the kidney dimensions acquired by the students were consistent with those acquired by the lecturers. Thus, the proposed kidney simulator can be used for the training of veterinary medicine students in ultrasonographic diagnostic techniques.
Training simulators can facilitate the acquisition and development of basic surgical skills, and they constitute a safe and humane method that does not harm animals in the process. The objective of this work was to creat...Training simulators can facilitate the acquisition and development of basic surgical skills, and they constitute a safe and humane method that does not harm animals in the process. The objective of this work was to create and evaluate a low-cost simulator to help undergraduate students of veterinary medicine acquire and practise vascular ligation skills. A training model was made by using easily accessible and inexpensive materials. Fifteen students, without prior surgical experience, each performed the orchiectomy technique on the simulator seven times. The emphasis was on performing vascular ligatures in both testicles of the simulator ( = 14), in two different scenarios: firstly, with the contents of the blood vessels not under pressure; and secondly, with the syringe plunger depressed by 1 ml to pressurise the blood vessels. The outcomes of the procedure in the simulator were based on three qualitative criteria: (no 'bleeding'), (light 'bleeding') and (heavy 'bleeding'). After the seventh attempt, all participants were able to perform vascular ligatures with at least a score of in both scenarios. By the 10th ligature attempt, they were all able to perform the procedure with a score of ( < 0.05). There was a trend toward a decrease in the time taken to carry out the procedure as learning progress was made during training, with this being significant from the 12th attempt ( < 0.05). The use of this low-cost simulator represents a useful didactic tool, which supports the acquisition and practise of manual skills by using methods that do not involve the use of animals. In addition, this training simulator could be useful in distance learning, in view of the ready accessibility of the materials required for its construction.
Echocardiography is an invaluable technique for the diagnosis of heart disease. The aim of this study was to develop 3-D models of healthy and diseased hearts of dogs and cats, and to evaluate their effectiveness in assi...Echocardiography is an invaluable technique for the diagnosis of heart disease. The aim of this study was to develop 3-D models of healthy and diseased hearts of dogs and cats, and to evaluate their effectiveness in assisting veterinary undergraduates to understand echocardiographic imaging planes. Resin models depicting the main echocardiographic imaging planes of normal hearts were created, as well as example hearts with features of mitral degeneration in dogs and hypertrophic cardiomyopathy in cats. After a theoretical class, fourth-year students were randomly assigned to one of two groups (model group or control group). The model group had access to the 3-D models, along with self-explanatory text about echocardiographic imaging planes; the control group only had access to the self-explanatory text. Both groups were allowed 2 weeks to study their allocated resources, after which the students undertook an assessment to evaluate their learning and completed a questionnaire about their experiences and satisfaction with the respective teaching method. A total of 39 students participated in the study, 19 in the model group and 20 in the control group. Students assigned to the model group spent more time studying ( = 0.0027). The proportion of students who achieved a satisfactory grade in the assessment was 89.5% in the model group and 60% in the control group ( = 0.0449). The 3-D models facilitated, and significantly improved, the identification of cardiac structures and disease-associated abnormalities, and the learning process in general. Additionally, the models seemed to provide greater student motivation for studying echocardiography.
Neuhaus W, Reininger-Gutmann B, Rinner B
… +62 more, Plasenzotti R, Wilflingseder D, De Kock J, Vanhaecke T, Rogiers V, Jírová D, Kejlová K, Knudsen LE, Nielsen RN, Kleuser B, Kral V, Thöne-Reineke C, Hartung T, Pallocca G, Leist M, Hippenstiel S, Lang A, Retter I, Krämer S, Jedlicka P, Ameli K, Fritsche E, Tigges J, Buettner M, Bleich A, Baumgart N, Baumgart J, Meinhardt MW, Spanagel R, Chourbaji S, Kränzlin B, Seeger B, von Köckritz-Blickwede M, Sánchez-Morgado JM, Galligioni V, Ruiz-Pérez D, Movia D, Prina-Mello A, Ahluwalia A, Chiono V, Gutleb AC, Schmit M, van Golen B, van Weereld L, Kienhuis A, van Oort E, van der Valk J, Smith A, Roszak J, Stępnik M, Sobańska Z, Olsson IAS, Franco NH, Sevastre B, Kandarova H, Capdevila S, Johansson J, Cederroth CR, Sandström J, Ragan I, Bubalo N, Spielmann H
Public awareness and discussion about animal experiments and replacement methods has greatly increased in recent years. The term 'the Three Rs', which stands for the Replacement, Reduction and Refinement of animal experi...Public awareness and discussion about animal experiments and replacement methods has greatly increased in recent years. The term 'the Three Rs', which stands for the Replacement, Reduction and Refinement of animal experiments, is inseparably linked in this context. A common goal within the Three Rs scientific community is to develop predictive non-animal models and to better integrate all available data from , and omics technologies into regulatory decision-making processes regarding, for example, the toxicity of chemicals, drugs or food ingredients. In addition, it is a general concern to implement (human) non-animal methods in basic research. Toward these efforts, there has been an ever-increasing number of Three Rs centres and platforms established over recent years - not only to develop novel methods, but also to disseminate knowledge and help to implement the Three Rs principles in policies and education. The adoption of on the protection of animals used for scientific purposes gave a strong impetus to the creation of Three Rs initiatives, in the form of centres and platforms. As the first of a series of papers, this article gives an overview of the European Three Rs centres and platforms, and their historical development. The subsequent articles, to be published over the course of 's 50th Anniversary year, will summarise the current focus and tasks as well as the future and the plans of the Three Rs centres and platforms. The Three Rs centres and platforms are very important points of contact and play an immense role in their respective countries as 'on the ground' facilitators of . They are also invaluable for the widespread dissemination of information and for promoting implementation of the Three Rs in general.