Evaluation of a gel-based versus micro-pellet diet for adult zebrafish (Danio rerio).

Ad libitum feeding of laboratory zebrafish has potential benefits for colony management, but would require a new type of diet, such as a gel that remains in the tank. We hypothesized that adult zebrafish fed a gel diet would have similar body size and reproductive success compared with those fed a standard micro-pellet diet. The gel diet's impact on water quality was determined to be safe for zebrafish prior to starting a 12-week feeding study. Two hundred adult AB zebrafish of mixed sex were randomly assigned to be fed exclusively either gel or micro-pellet diet. Fish body length and mass were measured every two weeks, and fish were bred within each feed group to assess fecundity. Zebrafish consumed less gel diet than expected. Body length, mass, and breeding success were lower in the gel diet fish than in the micro-pellet diet fish. Low consumption of the gel diet and/or nutritional differences between the two diets may have contributed to reduced growth and fecundity. Though the gel diet could reduce time personnel spend feeding and be safer for fish in static tanks, the tested formulation was not a satisfactory alternative to the control micro-pellet diet in a research zebrafish colony.

Experiences of early career researchers: Influences on the design and reporting of animal experiments, and the practical and emotional support needed to enhance best practice methods.

While best practice methodology in animal research aims to address reproducibility and translational issues, awareness and implementation remains low. Preclinical systematic reviews have highlighted many flaws, including issues with internal validity and reporting. With early career researchers (ECRs) heavily involved in all aspects of animal experiments, it is crucial we understand what shapes their research practices. Semi-structured interviews were conducted with 13 ECRs, including research masters, PhD and postdoctoral academics. Data were collected and analysed concurrently using constant comparison techniques and an iterative approach. Findings revealed low-level awareness of best practice recommendations but a desire to engage in dedicated workshops on designing and reporting animal experiments. Current laboratory practices and previous literature were main influences on research practice, more than institutional training. An unexpected finding was the discovery of ethical and emotional dilemmas ECRs faced when working with animals. This highlights the need for a multifaceted approach to better support junior researchers, both emotionally and practically, to encourage responsible science.

Guidelines for the establishment and functioning of Animal Ethics Commitees (Institutional Animal Care and Use Committees) in Africa.

Animals are used for scientific purposes across Africa to benefit humans, animals or the environment. Nonetheless, ethical and regulatory oversight remains limited in many parts of the continent. To strengthen this governance framework, the Pan-African Network for Laboratory Animal Science and Ethics brought together experts from 12 African countries to create an Africa-centric practical guide to facilitate the establishment and appropriate functioning of Institutional Animal Ethics Committees across Africa. The Guidelines are based on universal principles for the care and use of sentient animals for scientific purposes, with consideration of the cultural, religious, political and socio-economic diversity in Africa. They focus on 11 key elements, including responsibilities of institutions and of the Institutional Official; composition of the Committee; its responsibilities, functioning and authority; ethical application and review processes; oversight and monitoring of animal care and use and of training and competence; quality assurance; and the roles of other responsible parties. The intent is for African institutions to adopt and adapt the guidelines, aligning with existing national legislation and standards where relevant, thus ensuring incorporation into practice. More broadly, the Guidelines form an essential component of the growing discourse in Africa regarding moral considerations of, and appropriate standards for, the care and use of animals for scientific purposes. The increased establishment of appropriately functioning animal ethics committees and robust ethical review procedures across Africa will enhance research quality and culture, strengthen societal awareness of animals as sentient beings, improve animal well-being, bolster standards of animal care and use, and contribute to sustainable socio-economic development.

Eugenol as anesthetic for Cnesterodon decemmaculatus (Cyprinodontiformes, Poeciliidae).

Anesthetics are commonly used in fish for surgery and to facilitate capture, handling and transport in aquaculture and experimental procedures. In research, the selection of the anesthetic depends on its properties and on the recovery time. Eugenol has been pointed out as an effective anesthetic for fish, alternative to traditional drugs. Although Cnesterodon decemmaculatus is widely used as a model in ecological bioassays, no anesthetic protocol has been reported for this species. The aim of the present study was to evaluate the induction time (i.e. time to reach anesthetic stage VI) and recovery time in individuals of C. decemmaculatus subjected to eugenol at a fixed concentration, according to sex and pregnancy status. Forty-one fish were divided into three groups: males, pregnant females and non-pregnant females. They were measured for total length, standard length and weight, and the condition factor (K) was calculated. No significant differences in induction and recovery times were found for sex, pregnancy status and K between groups. Results are a contribution toward the development protocol of a standard anesthetic protocol for C. decemmaculatus.

Pain management in zebrafish : Report from a FELASA Working Group.

Empirical evidence suggests fishes meet the criteria for experiencing pain beyond a reasonable doubt and zebrafish are being increasingly used in studies of pain and nociception. Zebrafish are adopted across a wide range of experimental fields and their use is growing particularly in biomedical studies. Many laboratory procedures in zebrafish involve tissue damage and this may give rise to pain. Therefore, this FELASA Working Group reviewed the evidence for pain in zebrafish, the indicators used to assess pain and the impact of a range of drugs with pain-relieving properties. We report that there are several behavioural indicators that can be used to determine pain, including reduced activity, space use and distance travelled. Pain-relieving drugs prevent these responses, and we highlight the dose and administration route. To minimise or avoid pain, several refinements are suggested for common laboratory procedures. Finally, practical suggestions are made for the management and alleviation of pain in laboratory zebrafish, including recommendations for analgesia. Pain management is an important refinement in experimental animal use and so our report has the potential to improve zebrafish welfare during and after invasive procedures in laboratories across the globe.

The implementation of tunnel handling in a mouse breeding facility revealed strain-specific behavioural responses.

As a step towards implementing non-aversive handling techniques at a big mouse breeding facility in Germany, tunnel handling was introduced in a breeding unit comprising three inbred mouse strains. To assess whether tunnel handling would be feasible for the animal technicians in their everyday work and beneficial for the mice when being handled during weekly cage change only, the behaviour of tunnel- and tail-handled animals of both sexes was examined before, during and after the handling events over a period of nine weeks. Moreover, the time expenditure was compared between both handling techniques. It was possible to use the tunnel in all three mouse strains. However, the impact of the handling techniques on the behavioural parameters investigated in the present study were strain-specific. All behavioural parameters suggested that NZW mice benefited the most from tunnel handling. The results obtained from Hello Kitty and WNK mice were ambiguous, which may suggest that a brief handling session during the cage clean may have not been sufficient to habituate them to the process of handling. It took the animal technicians approximately 3 seconds longer per mouse when using a tunnel. The strain-specific results should encourage researchers to share their experiences with non-aversive handling techniques in different mouse strains, for example, along with their research articles.

Software for animal randomization: A tool for increasing the reproducibility of science.

Poorly designed preclinical studies may compromise human health due to erroneous conclusions regarding treatment effects in addition to contributing to experimental irreproducibility and wasted resources. Randomization is one of the crucial steps to enhance scientific rigor and is a commonly recognized bias-reducing instrument that increases the reliability and reproduction of studies involving animals (even with syngeneic animals). This procedure should be considered when planning a study and reported during data publication. In this context, this work aimed to highlight the importance of adopting quality measures in preclinical trials, with an emphasis on animal randomization. The 'Mouse Randomization' app was developed to help researchers estimate an adequate sample size to obtain significant statistical power, ensuring the ethical use of animals. This app is freely available on the internet to carry out animal randomization and calculate sample sizes for in vivo experiments. We believe that this brief discussion about animal randomization could raise awareness among researchers on how to improve the quality of preclinical research, increasing reproducibility and avoiding animal misuse.

A minimally invasive bronchoscopic approach for direct delivery to murine airways and application to models of pulmonary infection.

The laboratory mouse is used extensively for human disease modeling and preclinical therapeutic testing for efficacy, biodistribution, and toxicity. The variety of murine models available, and the ability to create new ones, eclipses all other species, but the size of mice and their organs create challenges for many in vivo studies. For pulmonary research, improved methods to access murine airways and lungs, and track substances administered to them, would be desirable. A nonsurgical endoscopic system with a camera, effectively a bronchoscope, coupled with a cryoimaging fluorescence microscopy technique to view the lungs in 3D, is described here that allows visualization of the procedure, including the anatomical location at which substances are instilled and fluorescence detection of those substances. We have applied it to bacterial infection studies to characterize better and optimize a chronic lung infection murine model in which we instill bacteria-laden agarose beads into the airways and lungs to extend the duration of the infection and inflammation. The use of the endoscope as guidance for placing a catheter into the airways is simple and quick, requiring only momentary sedation, and reduces post-procedural mortality compared with our previous instillation method that includes a trans-tracheal surgery. The endoscopic method improves speed and precision of delivery while reducing the stress on animals and the number of animals generated and used for experiments.

Evaluating and developing sealants for the prevention of pulmonary air leakage: A systematic review of animal models.

Sealants may provide a solution for pulmonary air leakage (PAL), but their clinical application is debatable. For sealant comparison, standardized animal models are lacking. This systematic review aims to assess methodology and quality of animal models for PAL and sealant evaluation. All animal models investigating lung sealing devices (e.g., staplers, glues, energy devices) to prevent or treat PAL were retrieved systematically from Embase, Pubmed and Web of science. Methodological study characteristics, risk of bias, reporting quality and publication bias were assessed. A total of 71 studies were included (N = 75 experiments, N = 1659 animals). Six different species and 18 strains were described; 92% of experiments used healthy animals, disease models were used in only six studies. Lesions to produce PAL were heterogenous, and only 11 studies used a previously reported technique, encompassing N = 5 unique lesions. Clinically relevant outcomes were used in the minority of studies (imaging 16%, air leak 10.7%, air leak duration 4%). Reporting quality was poor, but revealed an upward trend per decade. Overall, high risk of bias was present, and only 18.7% used a negative control group. All but one study without control groups claimed positive outcomes (95.8%), in contrast to 84.3% using positive or negative control groups, which also concluded equivocal, adverse or inconclusive outcomes. In conclusion, animal studies evaluating sealants for prevention of PAL are heterogenous and of poor reporting quality. Using negative control groups, disease models and quantifiable outcomes seem important to increase validity and relevance. Further research is needed to reach consensus for model development and standardization.

FELASA recommendations for the rehoming of animals used for scientific and educational purposes.

Directive 2010/63/EU of the European Parliament and the Council of 22 September 2010 states that at the end of a procedure, the most appropriate decision on the future of an animal previously used or intended for use in scientific procedures should be taken on the basis of animal welfare and potential risks to the environment. Member States may allow animals to be rehomed provided the health of the animal allows it, there is no danger to public health, animal health or the environment and if appropriate measures have been taken to safeguard the wellbeing of the animal. In countries where rehoming is permitted, it is the responsibility of the Animal Welfare Body to advise on a rehoming scheme which must include appropriate socialization in order to help facilitate successful rehoming, avoid unnecessary distress to the animals and guarantee public safety. This paper reviews the EU legislation, existing guidance, current literature and best practice to define rehoming, sets out general considerations for rehoming laboratory animals including socialization and provides practical advice on the steps required in a rehoming scheme. For those species most frequently rehomed, more detailed species-specific sections are included.

When less is more: Experimental Bishop-Koop technique for reduction in the use of laboratory animals for intestinal pathophysiological studies.

The use of animals to gain knowledge and understanding of diseases needs to be reduced and refined. In the field of intestinal research, because of the complexity of the gut immune system, living models testing is mandatory. Based on the 3Rs (replacement, reduction and refinement) principles, we aimed to developed and apply the derived-intestinal surgical procedure described by Bishop and Koop (BK) in rats to refine experimental gastrointestinal procedures and reduce the number of animals used for research employing two models of intestinal inflammation: intestinal ischemia-reperfusion injury and chemical-induced colitis. Our results show the feasibility of the application of the BK technique in rodents, with good success after surgical procedure in both small and large intestine (100% survival, clinical recovery and weight regain). A considerable reduction in the use of the number of rats in both intestinal inflammation models (80% in case of intestinal ischemia-reperfusion damage and 66.6% in chemical-induced colitis in our experimental design) was achieved. Compared with conventional experimental models described by various research groups, we report excellent reproducibility of intestinal damage and functionality, survival rate and clinical status of the animals when BK is applied.

Psychological stress and strain in laboratory animal professionals - a systematic review.

Laboratory animal professionals (LAP) are faced with various situations and tasks influencing their mental well-being. A systematic review has been conducted to investigate whether there are specific stressors for LAP and which moderators are relevant for the development of psychological strain. A comprehensive search following PRISMA Guidelines was carried out in June 2021. Results include 12 studies and have been summarized qualitatively in narrative synthesis and tabular presentation. Available literature indicates that LAP are facing stressors but does not allow for conclusions on specific stressful job duties other than euthanasia.Signs of strain are present in LAP. Specifically, participants in qualitative studies reported acute symptoms, while chronic manifestations were in focus in quantitative studies. Although a wide variety of moderating factors have been investigated, only social support has been rated as relevant by multiple qualitative and quantitative studies without contrasting results.According to current data, there is a risk for psychological strain in LAP. However, there is limited understanding of specific stressors and data on moderators is diverse. Further studies that focus on domain-specific knowledge and clearly distinguish stressors from moderators are necessary to set up institutional programmes addressing psychological strain in LAP.

Clean-catching urine from pigs: A method for collecting quality specimens for urinalysis and microbiological culturing in a laboratory environment.

Urine is an important biological specimen for assessing various metabolic functions and drug clearance. In urinary tract infection research, urine is particularly important as urinary bacterial titres constitute the main diagnostic outcome for assessing the course of infection. Collecting uncontaminated urine samples from pigs can be done by bladder catheterization or suprapubic bladder aspiration, which are both laborious and invasive procedures that require the need for anaesthesia. To improve animal welfare and optimize urine sampling protocols, we developed a method of clean-catching midstream urine specimens from pigs during spontaneous micturition. The quality of urine specimens collected by clean-catch, bladder catheter and suprapubic bladder aspiration were compared using microbiological culturing. We show that urine specimens collected by clean-catch are only minimally contaminated by skin- and vaginal-commensals with no influence on urinary bacterial titres during ongoing infection. In conclusion, we describe a method in which spontaneous micturition can be prompted in pigs, facilitating fast and reliable collection of quality specimens suitable for microbiological culturing. The method supersedes the need for anaesthesia, which not only represents a considerable refinement in terms of animal welfare but also facilitates more frequent collection of specimens that can enhance the scientific outcome of experimental animal studies in pigs.

A systematic review on reporting of refinement measures in mouse ECG telemetry implantation surgery.

Telemetric monitoring is used in many scientific fields, such as cardiovascular research, neurology, endocrinology, as well as animal welfare research. Nowadays, implanted electrocardiogram (ECG) radiotelemetry units are the gold standard for monitoring ECG traces, heart rate and heart rate variability in freely moving mice. Telemetry technology can be a valuable tool when studies utilize it adequately, while prioritizing animal welfare. Recently, concerns have been raised in many research fields, including animal research, regarding the reproducibility of research findings, with insufficient reporting being one of the underlying causes.A systematic review was performed by making use of three literature databases, in order to include all publications until 31.12.2019, where the surgical placing of ECG recording telemetry devices in adult mice was involved. Data extracted from the publications included selected items recommended by the ARRIVE guidelines. We focused on aspects related to the refinement of the surgery and experimental conditions that aim to improve animal welfare. In general, the quality of reporting was low in the analyzed 234 publications. Based on our analyses, we assume there has been no improvement in this field's reporting quality since 2010 when the ARRIVE guidelines on reporting were introduced. Additionally, even though expert recommendations on telemetry surgery refinement have been available since many years now, no increase in uptake (or reporting) of these measures prior (e.g., acclimatization), during (e.g., asepsis) or after (e.g., social housing) the surgery could be observed.

Islam and Veterinary Science: Rethinking Animal Suffering Through Islamic Animal Ethics and the Evolving Definition of Halal Slaughter.

In the midst of recent European activism against religious slaughter, the idea that religious slaughter is cruel to animals is often seen as commonsense, and the mandatory pre-slaughter stunning is often portrayed as the moral technology that assures animal welfare. Nevertheless, this portrayal seems to blur the fact that the current notion of animal welfare itself is built upon a changing selection of value assumptions, which are not without problems or academic debates. It also ignores the fact that contemporary veterinary scientists and Muslim scholars have been working together for four decades to learn more about farm animals and their suffering. Despite stereotypes, the idea of animal ethics is not foreign to Islam. In Islam, animals represent God's wisdom and wonder, and humans are obliged to attend to their health and living conditions. When killing animals for food is conducted, the slaughter must be done in the name of God as a sacred ritual in order to assure that the life of the animal is not taken lightly and that the slaughter is not a sign of hostility toward the universe. Before the act of sacrifice, the animal must be healthy, and no harm should be forced upon it. Accordingly, the requirement of pre-slaughter stunning has posed a question to Muslim scholars: Does stunning kill the animal or cause harm? What defines harm, and whose definition counts? This paper reconstructs a socio-technological history of halal slaughter through scientific research on animal suffering since the 1980s. On the basis of archival research of New Zealand veterinary scientists' works and in-depth interviews with Malaysian veterinary scientists, this article outlines three phases of the evolution of halal slaughter that aims to fulfill multiple sets of moral obligations toward farm animals, and demonstrates how veterinary scientists establish common ground between secular and Islamic animal ethics. In this vein, I am envisioning a possibility of veterinary anthropology that recognizes the field's trans-cultural characteristics, and continues to challenge the rigid binaries between the West and the Rest, and between science and culture.

An Ethics Toolkit to Support Animal-Centered Research and Design.

Designers and researchers who work with animals need to employ an array of ethical competencies to guarantee the welfare of animals taking part in animal-centered research. The emerging field of Animal-Computer Interaction (ACI), which deals with the design of animal-centered interactive systems, considers ethics a fundamental concern when working with animals, and ACI researchers have proposed ethics frameworks in response to these concerns. Ethical approaches proposed within the field tend to be normative but, on their own, norms may not be sufficient to support designers who will inevitably face unexpected and ethically charged situations as the research progresses. During a research project, focused on the design of dog-friendly controls for Mobility Assistance Dogs (MADs), these limitations came to the fore. Drawing from situated ethics approaches, developed to support researchers' ethical engagement with vulnerable populations such as children and differently abled adults, this paper presents an ethics toolkit that aims to support animal-centered research and design by enabling researchers to make ethically sound situated decisions as their work progresses. The toolkit comprises three templates, each of which asks a series of questions aiming to articulate the ethical baselines of individual team members and of their research project, and to inform the development of a series of ethical guiding statements to better prepare designers to make ethical situated decisions. The application of the toolkit during the research with MADs helped the field researcher to clearly and systematically articulate the project's ethos and understand the ethical stance that guided the research team's interactions with the dogs, their trainers, and their human partners throughout the project. It also fostered a practice of active reflection within the team, which helped them to maintain their commitment to the project's ethos in the face of unexpected ethical challenges. We propose that, beyond supporting ACI research, the toolkit could support the ethical engagement of researchers and practitioners who work for and with animals in many other settings.

The use of ball pits and playpens in laboratory Lister Hooded male rats induces ultrasonic vocalisations indicating a more positive affective state and can reduce the welfare impacts of aversive procedures.

The advancement and quality of science rely on research that is robust and unbiased in its experimental design, execution, analysis, and reproducibility. In preclinical research, a better understanding of animal emotions and refinement of their husbandry, housing, and handling are important goals in providing good animal welfare in a laboratory setting which underpins rigorous research quality. Induction of positive emotional state in animals is a key component of their well-being, and one approach is to increase their environmental complexity using, for example, ball pits or playpens in rats. In this study, we recorded 50 kHz ultrasonic vocalisations (USVs) during animals' exposure to the ball pit and playpen. We have previously shown that 50 kHz USVs provide a graded and quantifiable measure of an animal's emotional state, and here find that access to the ball pit and playpen increases 50 kHz USVs, indicative of a more positive affective state. Using our affective bias test (ABT) we next quantified the animals' emotional response to an aversive intervention and whether this could be attenuated by access to a playpen. The playpen exposure completely mitigated the negative affective state induced by an anxiogenic drug when compared with animals who experienced the drug in the home cage. Together, these findings suggest ball pits and playpens provide a simple and effective method to improve the welfare of laboratory rats and reduce the cumulative suffering they experience from their housing conditions and minor, aversive procedures.

Personality traits and meat consumption: The mediating role of animal-related ethical concerns.

Prior research suggests that personality traits are associated with meat consumption. However, this association is not uniform across all types of meat. For instance, Big Five personality traits such as openness and agreeableness are negatively associated with red meat consumption but positively associated with fish. Using a large sample of Chilean university students (N = 1,149), we examined whether these differential meat consumption patterns can be explained by an intermediary variable of animal-related ethical values. Structural equation modeling was employed to test the hypothesized associations. The results suggest that animal-related ethical values mediate the effect of certain personality traits on the consumption of beef and poultry.

Ultrasound-guided external jugular and femoral arterial cannulation for juvenile swine.

Central venous and arterial access through minimally invasive techniques has been described in adult pigs. This article demonstrates success in juvenile animals. Using ultrasound guidance and the modified Seldinger technique, 5 Fr/15 cm single-lumen central venous catheters and 20 Ga 4.5 cm femoral arterial catheters were placed in six Yorkshire cross-bred swine. All six cases had no loss of venous catheter patency or infection during the 96-hour follow-up period. Arterial catheters remained patent, and no significant bleeding was noted after removal.

CORE management: The reality principle.

Responsible science is mandatory for every research institution. As economic challenges, fast evolving technologies and competitiveness impact drastically the scientific productivity and quality, institutions must take action. Research core facilities using animal models (CORE) are central in biomedical institutions. Therefore, building a strong CORE represents a priority for research organizations. More precisely, COREs must define their purpose, ensure proper long term resources and promote ethics and transparency. The heads of COREs play, as managers, a key role in the development and in the coordination of all activities. They deal with multiple challenges, such as divergent objectives, heavy workload and limited resources, exposing them to psychosocial risks, and might compromise their ability to react rationally to the pressure. The implementation of a culture of care and of social responsibility is essential for COREs and for their institutions. In this, the collaborative efforts between institutions' officials, administrative staff and scientists allow the support of CORE decisions, the development of innovative tools and the promotion of a responsible science. Overall, the improvement of the CORE strategy and functioning results in increased scientific success rates for the institution, while reducing psychosocial risks for personnel.