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.

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.

Regular health monitoring is crucial in laboratory animal facilities to determine the presence or absence of specific pathogens. One common approach to monitoring involves the use of sentinel animals, which are periodically exposed to biological material from the cages being monitored. At a certain point, some of these sentinel animals are tested for pathogens. This article discusses designing an effective sampling scheme to meet desired quality standards. It addresses questions such as the number of sentinel animals required, the frequency of sampling biological material, the selection of cages based on facility set-up, and the optimal frequency and quantity of sentinel animal tests. While existing design formulas are available for simple random sampling, no quantitative recommendation exists for using sentinel animals to the best of our knowledge. We propose a Monte Carlo simulation-based approach in this article to address this. Our algorithm has been implemented in a publicly accessible web page at http://nolan.cnb.csic.es/sentinelcagesmanager.

Pilots are small-scale initial experiments that are intended to guide the design of future, larger studies, with a view to increasing their effectiveness. In this statistical primer we highlight five common mistakes that limit the utility of pilot studies and provide practical guidance to avoid such errors and increase their effectiveness. The common thread connecting these mistakes is insufficient planning and over-interpretation of the results. This approach compromises the ultimate goals of the research programme and the future experimental cascade. In support of our view that over-interpretation is an error, we present a simple simulation to demonstrate that pilots will generally generate an inaccurate estimate of the variability of the biological endpoint under study and that frequent under-estimation will lead to inconclusive and unethical subsequent experiments. We argue that well planned pilots are an important part of the research cascade and still need to be implemented to a high standard.

Niemann-Pick disease type C (NPC) is a lethal genetic disease with mutations in NPC1 or NPC2 gene. Npc1-deficient (Npc1-/-) mice have been used as a model for NPC pathogenesis to develop novel therapies for NPC. However, Npc1-/- mice are infertile; thus, securing sufficient numbers for translational research is difficult. Hence, we attempted reproductive engineering techniques such as in vitro fertilization (IVF) and sperm cryopreservation. For the first time, we succeeded in producing fertilized oocytes via IVF using male and female Npc1-/- mice. Fertilized oocytes were also obtained via IVF using cryopreserved sperm from Npc1-/- mice. The obtained fertilized oocytes normally developed into live pups via embryo transfer, and they eventually exhibited NPC pathogenesis. These findings are useful for generating an efficient breeding system that overcomes the reproductive challenges of Npc1-/- mice and will contribute to developing novel therapeutic methods using NPC model mice.

Kidney cysts in humans are mainly caused by inheritable polycystic kidney disease. Although they are a regular finding in laboratory mice, their occurrence upon dissection has not been systematically investigated, yet. Therefore, the aim of this report was to investigate on prevalence, phenotype and aetiology of spontaneously occurring kidney cysts in mice by retrospectively analysing the laboratory-receipt tables of the in-house laboratory of a central animal facility in North Rhine-Westphalia, Germany, years 2009-2019. A percentage of 0.4% of dissected mice displayed kidney cysts, with more male than female animals affected and average age equal to that of all dissected animals. Preliminary report in half of the cases was distended abdomen, and a few individuals displayed additional pathologic alterations of kidneys, most commonly dilated renal pelvis, or extrarenal comorbidities. Kidney cysts occurred independently of a renal phenotype of the transgenic strain or presence of infectious agents in health monitoring. To conclude, kidney cysts were characterized as harmless for affected mice but, as inheritability is suggested according with the literature, affected animals should be excluded from breeding.

The aim was to determine changes in clinical parameters, glucose concentration, cortisol and behavior in colony queens in no music conditions compared with exposing to different genres of music. Mixed breed clinically healthy queens (N = 9) were used. Queens were studied under no music conditions (control=CON) and auditory enrichment: Soft Rock (M1), Motown (M2), Pop (M3), Frenchcore (M4) and music that was composed to be species-appropriate for cats (M5). The queens underwent auditory enrichment, including three days of silence (D1-3), five consecutive days of auditory enrichment (D4-8) and three days of silence (D9-11). We recorded clinical parameters, glucose, cortisol and behavior. Data were analyzed with GLIMMIX. Queens showed a significant increase of femoral pulse during exposure to M1, M2 and M4; and respiratory rate during exposure to M1, M2, M3 and M4 compared with CON. There was a significant increase in glucose during exposure to M2 and M5 compared with CON. However, there was a significant decrease in glucose during exposure to M4 compared with CON. There was a significant decrease in cortisol during exposure to M2 and M5 compared with CON. When we evaluated the ethogram, we observed a significant decrease in the percentage of interaction with other cats in M1 and M2 compared with CON. In addition, we found a significant decrease in the purring in M1 and M3 compared with CON. Auditory enrichment can be beneficial in situations that cause discomfort and distress in colony cats, such as in feline hospitalization; however, it should be acknowledged that there are limits to direct extrapolation.

The intranasal route enables direct delivery of multiple substances from the nose to the brain, through olfactory and trigeminal pathways, bypassing the blood-brain barrier and avoiding systemic absorption. Despite the potential of this route, the various administration approaches make data reproducibility and interpretation challenging, emphasizing the necessity to establish a consistent methodology. Considering this, the aim of our study was to assess and compare the distribution of two dye volumes (30 µl and 50 µl) in the nasal cavity of rat cadavers. We employed three distinct methods of intranasal delivery: nose drops, by pipette tip, or cannula inserted into the nasal cavity. The results indicated that for both volumes, using the nose drops and the pipette tip methods, the dye dispersion occurred mainly in the vestibule, respiratory and olfactory regions, without reaching the olfactory bulbs. Using the cannula method, the deposition predominantly occurred in the respiratory and olfactory regions, with the dye reaching 66.7% and 100% of the olfactory bulbs, respectively, to low and high volume. Furthermore, the results demonstrated differences between the two volumes, in the pharynx, larynx, trachea, septal window, and incisive papilla, where an increased dye presence was observed with the 50 µl instillation across all three methods. According to our results, the intranasal delivery with a cannula was the most effective method for dye deposition in the olfactory region. However, further studies in live animals will be necessary to determine and refine the administration method that consistently allows specific deposition in the olfactory system.

This prospective anatomical study aimed to establish an ultrasound-guided technique to the quadratus lumborum (QL) plane in sheep cadavers. Thirteen cadavers, weighing less than 117 kg, were included. In phase 1, one cadaver underwent dissection and two cadavers underwent 3D computed tomographic reconstruction for anatomical evaluation of the thoracolumbar region. In phase 2, two cadavers were used to compare two ultrasound techniques to the QL plane: lateral to the QL muscle with a transversal approach (LQL) and transmuscular between QL and psoas muscles with a longitudinal approach (TQL). For LQL, the reference was the first lumbar transverse process, whereas for TQL, it was the intertransverse region between the first and second lumbar vertebrae. The needle was advanced in-plane towards the specific target for each technique and a total of four injections were performed using 0.4 ml kg-1 of a dye-lidocaine solution. In phase 3, 10 cadavers received bilateral LQL injections (n = 20). All cadavers were then dissected to evaluate spread of dye. In phase 2, following LQL injections, no dye was observed in undesired locations; however, the dye was noted in the retroperitoneal space (1/2) after TQL injections. In phase 3, the 13th thoracic, first, second, third lumbar nerves, and sympathetic trunk segments were stained in 80%, 95%, 100%, 45% and 35% of the injections, respectively. In conclusion, the LQL technique was feasible, allowing staining of the spinal nerves innervating the cranial abdomen in sheep cadavers. Further studies in live animals are warranted.

Male Zucker Diabetic Sprague Dawley rats were housed in standard individually ventilated cages with floor area of 1500 cm2, and were placed in a 'playcage' (a large open cage) for three visits per week from the age of 16-18 weeks. The playcage was introduced in an attempt to reverse the compulsive behaviour that the rats displayed in the individually ventilated cages, with the purpose to increase their well-being and to provide cognitive as well as physical stimulation. After two weeks of periodic stays in the playcage, the rats' repetitive behaviour in their home cage ceased, and the rats displayed signs of happiness and excitement when they were in the playcage. The observations strongly indicate that periodic stays in a larger playcage can be an alternative environmental enrichment for laboratory rats when housing in a larger home cage is not an option.