The crystal structure of amesite-2H1 from the Postmasburg Mn ores of South Africa has been refined to R = 4.7% using Siemens P4 rotating-anode MoKα X-ray data and SHELXL-93 software. Partial ordering of tetrahedral Si,Al and especially of octahedral Mg,Al has reduced the ideal P63cm symmetry to triclinic P1 (refined as C1). Tetrahedral rotation of 15° moves the basal oxygens toward the octahedral cations in the same layer but away from the OH H-donors of the next layers. The ordering pattern of AlIV and AlVI is unique in that the locus of Al-rich sites consists of 2 intersecting sets of zigzag lines parallel to X2 and X3, plus 2 Al-rich octahedra in layer 2 instead of the usual 1 site.

Nacrite crystals from a vug within a matrix of dickite at Red Mountain near Silverton, Colorado, have a = 8.906(2), b = 5.146(1), c = 15.664(3) Å, β = 113.58(3)°, V = 657.9(3) Å3, and space group Cc. The structure was solved by direct methods to determine phase angles, followed by electron density maps to locate all atoms. Refinement by least-squares ceased at R = 4.5%. Each 7 Å layer has structural detail very similar to those of dickite and kaolinite, although nacrite stacking is based on — a/3 interlayer shifts along the 8.9 Å axis (with octahedral cations alternating between the I and II sites in successive layers), whereas dickite and kaolinite are based on shifts of — a/3 along the 5.1 Å axis (with octahedral cations in the same set of sites in each layer). The angle of tetrahedral rotation is 7.8°, and the octahedral counterrotations are 7.6° and 8.1°. The H+ protons were located on DED maps. The inner 0..H1 vector points exactly toward the vacant octahedron and is depressed — 18.6° away from the level of the octahedral cations. All three surface OH groups have 0...H vectors at 50° to 66° to (001), although OH2 may not participate in interlayer hydrogen bonding. All three interlayer OH-H-O contacts are bent to angles between 132° and 141° and form contacts between 2.94 and 3.12 Å. The interlayer separation of 2.915 Å is slightly larger than in dickite, interpreted as due to a less favorable meshing of the oxygen and hydroxyl surfaces in nacrite—a direct consequence of layer shifts along the 8.9 Å axis.

Primates are important species for biomedical research and ensuring their good welfare is critical for research translatability and ethical responsibility. Systematic animal welfare assessments can support continuous programme improvements and build institutional awareness of areas requiring more attention. A multi-facility, collaborative project aimed to develop and implement a novel primate welfare assessment tool (PWAT) for use with research macaques. PWAT development involved: establishing an internal focus group of primate subject matter experts, identifying animal welfare categories and descriptors based on literature review, developing a preliminary tool, beta-testing the tool to ensure practicality and final consensus on descriptors, finalising the tool in a database with semi-automated data analysis, and delivering the tool to 13 sites across four countries. The tool uses input- and outcome-based measures from six categories: physical, behavioural, training, environmental, procedural, and culture of care. The final tool has 133 descriptors weighted based upon welfare impact, and is split into three forms for ease of use (room level, site level, and personnel interviews). The PWAT was trialled across facilities in March and September 2022 for benchmarking current macaque behavioural management programmes. The tool successfully distinguished strengths and challenges at the facility level and across sites. Following this benchmarking, the tool is being applied semi-annually to assess and monitor progress in behavioural management programmes. The development process of the PWAT demonstrates that evidence-based assessment tools can be developed through collaboration and consensus building, which are important for uptake and applicability, and ultimately for promoting global improvements in research macaque welfare.

Factors controlling the crystal structure of phlogopite have been widely investigated; but the role of electrostatic interactions, for example, has received much less attention than other factors. The purpose of the present study was to peform a single-crystal refinement of an Al-saturated phlogopite and to use that refinement to supplement crystal-chemical analyses. The phlogopite investigated was from the Rumford quadrangle, Maine, and has the following chemistry: ${\left({\text{K}}_{0.81}{\text{Na}}_{0.03}\right)}_{\Sigma =0.84}{\left({\text{Mg}}_{1.84}{\text{Fe}}_{0.52}^{2+}{\text{Al}}_{0.45}{\text{Mn}}_{0.02}{\text{Ti}}_{0.07}^{4+}\right)}_{\Sigma =2.90}{\left({\text{Si}}_{2.78}{\text{Al}}_{1.22}\right)}_{\Sigma =4}{\text{O}}_{10}{\left({\text{OH}}_{1.81},{\text{F}}_{0.05}\right)}_{\Sigma =1.86}$\$\end{document}. The sample is a 1M polytype with C2/m symmetry and cell dimensions of a = 5.3220(4), b = 9.2170(7), c = 10.2511(8) Å, and β = 100.081(1)°. Hydrogen atoms were located and the crystal structure was refined to give parameters R1 = 0.0301 and weighted R2 = 0.0887. The octahedral M1 site was larger than the M2 (average M1—O: 2.079 Å, average M2—O: 2.062 Å) and the electron counts were equal (M1 = M2 = 14.8 e−); based on bond distances, which are more accurate than electron counts in determining occupancy; this result is consistent with a slight preference of Mg for M2 and Fe2+ for M1.

Thirty-five Al-rich, natural phlogopite-1M samples that are of (1) high metamorphic grade, and that have (2) total Al contents ⩾ 1.27 atoms per formula unit (a.p.f.u.), (3) Fe3+ contents ⩽ 0.11 a.p.f.u., and (4) Mn contents ⩽0.10 a.p.f.u. along with the newly described phlogopite, exhibited crystal chemical trends related to increasing Al content. Octahedral substitutions of smaller, high-charge cations (i.e. Al) apparently decrease distortions in the octahedral sites and produce longer M2—O4 distances. In addition, VIFe-F avoidance apparently occurs in high Al-content samples, which are generally high in VIFe. The data set also shows that these samples have limited ordering among M sites (Fe2+ in M1 and Al in M2), an increase in β (99.96° to 100.32°) possibly caused by cation ordering and therefore size differences of M1 and M2, and interlayer (A) sites with A—Oouter distances that increase and A—Oinner distances that decrease with increasing Ti content.

Computer models were used to simulate electrostatic interactions in phlogopite structures with variable Al concentrations utilizing Pauling’s electrostatic valency principle, which considers first-coordination electrostatic interactions. The model results were compared to the maximum Al concentrations in natural and synthetic phlogopite samples. Model results revealed no indications (e.g. a limit reached or a sudden change occurred) that charge saturation/undersaturation of the apical oxygen atoms at Al contents equal to the maximum in natural and/or synthetic samples causes instability that could not be balanced by bond-length variation. However, a cation of higher charge substituting at M1 (or M2) may result in higher electrostatic repulsions between the other octahedral sites. Thus, the Al3+ content in the octahedral sites may reach a maximum, with Fe2+ for Mg substitutions favored.

Experimental procedures involving farm animals are often associated with stress due to restraining. Stress can be reduced through use of positive reinforcement training, which then serves as refinement according to the 3Rs principles. Trainer skills, however, may influence the feasibility and success of animal training. The potential influence of trainer skills as well as the education of animal trainers are rarely described in literature but are necessary information for the implementation of positive reinforcement training as a refinement measure. To investigate the effect of educational programmes on animal trainers, we compared the training success of two groups of participants in training goats to elicit a behaviour that would allow simulated venipuncture. One group was educated in a two-day workshop while the other was provided with specific literature for self-instructed learning. Training success was evaluated using an assessment protocol developed for this study. A greater training success in the WORKSHOP GROUP, reflected by objective and subjective measures, was clearly supported statistically. In addition, 73 versus only 13% of the participants of the WORKSHOP GROUP and the self-instructed BOOK GROUP, respectively, stated that they could completely implement the knowledge gained in the course of this study. Our results indicate that more intensively educated trainers can train animals more successfully. In conclusion, if animal training is implemented as refinement, animal caretakers should receive instruction for positive reinforcement training.

In 2019, Andrews and Newman [‘Partitions and the minimal excludant’, Ann. Comb. 23(2) (2019), 249–254] introduced the arithmetic function $\sigma \textrm {mex}(n)$, which denotes the sum of minimal excludants over all the partitions of n. Baruah et al. [‘A refinement of a result of Andrews and Newman on the sum of minimal excludants’, Ramanujan J., to appear] showed that the sum of minimal excludants over all the partitions of n is the same as the number of partition pairs of n into distinct parts. They proved three congruences modulo $4$ and $8$ for two functions appearing in this refinement and conjectured two further congruences modulo $8$ and $16$. We confirm these two conjectures by using q-series manipulations and modular forms.

Training using positive reinforcement is increasingly recognised as a valuable tool for the humane and effective management and use of laboratory-housed non-human primates. We utilised a mixed-mode questionnaire to survey use of training and other learning processes (socialisation, habituation and desensitisation) in over half of UK establishments using and breeding primates. The survey demonstrated that there is widespread awareness of training as a refinement technique and appreciation of its diverse benefits, but training is not used as widely or as fully as it might be. This is due to real constraints (principally staff and time and a lack of confidence in ability to train), and perceived constraints (such as a supposed lack of published information on how to train and assessment of the benefits, and an overestimation of the time investment needed). There is also considerable variation between establishments in the purposes of training and techniques used, with a reliance on negative reinforcement in some. We conclude that there is opportunity for refinement of common scientific, veterinary and husbandry procedures (such as blood and urine collection, injection, capture from the group and weighing) through use of positive reinforcement training, especially when combined with appropriate socialisation, habituation and desensitisation. We end this paper with recommendations on best practice, training techniques and staff education.

Russell and Burch's Three Rs principle of replacement, reduction and refinement offers a useful concept for the scientific and ethical evaluation of the use of animals in scientific procedures. Replacement, reduction and refinement are often considered separately, but when applied, one of the Three Rs may have a positive or negative effect on one or both of the other Rs. This paper explores the interplay between the Three Rs and provides examples where the Three Rs have a positive interaction and where they are in conflict with each other. For example, all Three Rs positively interact in the use of cell cultures, but validation studies of replacement techniques may initially increase the numbers of animals used; therefore replacement and reduction are in conflict. Several models of cost-benefit analyses, used by animal ethics committees to justify or reject animal experimentation, contain elements such as quality and significance of the research, the credibility of the research group and the discomfort caused to the animals. Although these models consider the Three Rs, each R is considered independently of the others. Consequently, moral dilemmas may arise when reviewing proposals in which the Three Rs conflict. Currently there is no legal guidance relating to the prioritisation of the Three Rs, but guidance is required to facilitate their use. For example, does a significant reduction in animal numbers justify increased individual suffering? Moral justifications deserve more attention when considering the Three Rs in general, and when considering the application of one or more Rs to a procedure, to a protocol, or to the wider research programme.

There is an ethical and scientific need to minimise the harm experienced by animals used in scientific procedures and to maximise their well-being. Welfare can be improved by the refinement of practice, particularly if these refinements are applied to every aspect of the life of an animal used in the laboratory, from birth to death. Primates are considered likely to have a greater capacity for suffering than other sentient species and therefore refinement of their use is particularly important. The refinement of the human impact on laboratory-housed primates and of housing and husbandry practices are dealt with in parts I and II of this three-part review. In part III, methods of refinement that can be applied specifically to the use of primates in procedures, are summarised and discussed, together with a description of some current practices, and the scientific evidence that suggests that they should no longer be used. Methods of refinement of identification, capture and restraint, sampling, administration of substances, humane endpoints, and euthanasia are included. If these methods are used, taking into account species-specific differences and needs, it is concluded that harm can be minimised and primate welfare improved.

In order to safeguard the welfare of laboratory-housed non-human primates, refinement techniques should be applied to every aspect of the life of animals used in the laboratory, from birth to death, with the aim of both minimising harm and maximising well-being. In this second part of a three-part review we summarise published information on housing and husbandry practices, and describe ways to minimise contingent inhumanity associated with the use of primates in laboratories and their breeding and supply (where inhumanity is defined as the infliction of distress). We also discuss methods by which the welfare of these animals can be maximised on a day-to-day basis. The principles of enrichment, aspects of the housing environment, social and physical enrichment and acquisition are discussed. Refinement of the influence of humans and experimental procedures are discussed in Parts I and III of this review, respectively.

The welfare of non-human primates used in scientific research must be safeguarded to promote scientific validity and for ethical reasons. Welfare can be improved by the refinement of practice, particularly if these refinements are applied to every aspect of the life of an animal used in the laboratory, from birth to death with the aim of both minimising harm and maximising well-being. Many refinement methods have been described in nationally and internationally accepted guidelines on laboratory practice, but awareness of these guidelines is not universal. In Part I of this review, we examine the influence of humans on non-human primates and summarise and evaluate methods of refinement that are or could be used to reduce suffering and improve welfare. In particular we focus on staff selection, education and training, human–animal bonds, staff communication, and training primates. In Parts II and III, refinements of housing, husbandry and experimental procedures are reviewed.

Refinement techniques are being increasingly employed in all fields of animal research to try to ensure that the highest standards of welfare are upheld. This review concerns one of the main emerging techniques for the assessment of welfare itself, namely the non-invasive measurement of glucocorticoids (GCs) as indicators of stress. The paper is divided into three sections. The first discusses the relationship between GCs and stress. The second section considers whether factors other than stress are linked to rises in GCs, eg exercise, oestrus cycle and diet. The final part examines the reliability of the non-invasive techniques that measure GCs from samples of saliva and faeces. Although it is important to take into account some caveats associated with the methodologies employed, it is concluded, nevertheless, that these techniques can give accurate and reliable information regarding the welfare status of an individual or group of animals without the procedures themselves causing any kind of distress to the subjects.

Russell and Burch's Three Rs (replacement, reduction and refinement) remain the cornerstone for principles guiding humane experimental research. However, the concept of refinement has evolved considerably since its first inception and there have been numerous interpretations, some of which are regressive from the original definition. In this paper we examine the interpretations of refinement, and propose a harmonised progressive definition that is in line with changes in animal ethics and animal welfare science. Our definition should be applied to all aspects of refinement: those related to housing, husbandry and care, techniques used in scientific procedures, periprocedural care, health and welfare monitoring, and experimental design. We argue not only that the concept should include the avoidance or minimisation of adverse effects experienced at any time during the life of an animal destined for use in a laboratory, but also that it should be applied to the founder animals. Furthermore, we take a proactive stance and argue that refinement should include enhancing well-being through environmental enrichment. The acceptance and application of this new definition by legislative authorities and in guidelines would represent a significant step forward for animal welfare.

Mongolian gerbils (Meriones unguiculatus) are a widely used model species in parasitology and epilepsy research. Under standard laboratory housing conditions gerbils develop stereotypic digging, a behavioural abnormality, which is caused by the lack of a burrow in the laboratory cage. Here we show that gerbils reared with access to an opaque artificial burrow (nest-box with access tube) develop significantly less stereotypic digging than those reared with transparent artificial burrows. Subadult gerbils also preferred opaque artificial burrows to transparent ones. Based on these findings we developed an artificial burrow system that could be integrated into laboratory Makrolon Type IV cages to prevent the development of stereotypic digging in gerbils by addressing their behavioural needs. Faecal cortisol levels were measured as a non-invasive method of comparing stress reactions in gerbils given access to the new integrated artificial burrow system or to an equivalent transparent burrow. Behavioural differences were observed between gerbils in the two housing conditions, but faecal cortisol levels were unaffected. We conclude that simple refinement of housing is effective in improving gerbil welfare in the laboratory.

A survey was undertaken to evaluate how animal discomfort, pain, suffering and distress are recognised and assessed in UK scientific procedure establishments. In total, twenty-eight establishments were visited between 1999 and 2001 and 137 people participated, including animal technicians, veterinarians and scientists. The full results, conclusions and recommendations of the survey have been published elsewhere (Hawkins 2002). The study showed that people are concerned about animal suffering, want to be able to prevent and alleviate it, and are aware that there are a number of practical problems that need to be overcome. These include animals concealing clinical signs, which leads to difficulties in detecting incipient discomfort and distress, and human subjectivity when assessing animals. The clinical signs used as indicators of potential pain, suffering and distress are largely subjective. Participants at all establishments agree that a ‘team’ approach to animal monitoring is the best way to ensure consistency and effectiveness. All twenty-eight establishments use clinical observation sheets to assist with animal assessment and monitoring, nine also use score sheets and seven use computerised data management systems. This paper concludes with recommendations based on the survey findings, with respect to monitoring techniques, assessment protocols and training issues, which aim to facilitate more effective animal assessment and monitoring.

Buprenorphine is a widely used analgesic for laboratory rodents. Administration of the drug in a desirable food item for voluntary ingestion is an attractive way to administer the drug non-invasively. However, it is vital that the animals ingest the buprenorphine-food-item mix as desired. The present study investigated how readily female and male mice (Mus musculus) of two different strains consumed buprenorphine mixed in a commercially available nut paste (Nutella®), and whether variation between genders and strains would affect the subsequent serum concentrations of buprenorphine. Buprenorphine at different concentrations mixed in Nutella® was given to male and female C57BL/6 and BALB/c mice in a complete cross-over study. Pure Nutella® or buprenorphine (1.0-3.0 mg kg−1 bodyweight [bw]) mixed in 10 g kg−1 bw Nutella® were given to the mice at 1500h. The mice were video recorded until the next morning, when blood was collected by submandibular venipuncture. The concentration of buprenorphine in the Nutella® mix did not affect the duration of ingestion in any of the groups. However, female mice consumed the Nutella® significantly faster than males. Repeated exposure significantly reduced the start time of voluntary ingestion, but not the duration of eating the mixture. These differences did not however affect the serum concentration of buprenorphine measured 17 h post administration.

In mouse (Mus musculus) models of diabetic nephropathy (DN), one of the most important read-outs is the 24-h urinary albumin excretion (UAE). The 24-h urine collection is usually performed by single housing mice in metabolic cages on wire mesh without enrichment. This is known to be stressful for the mice. Therefore, it was investigated if shorter urine collections would be sufficient to get reliable assessments of albuminuria. Twenty-one diabetic (C57BLKS-Leprdb/ db) and ten non-diabetic mice (C57BLKS-Leprdb/+) were placed in metabolic cages at 15 and 20 weeks of age (WoA) for 24 h. Urine samples were taken at 4, 6, 18 and 24 h and albumin and creatinine concentration were measured. Four- and 6-h UAE was found to correlate significantly with 24-h UAE. Furthermore, a significant correlation was found between 24-h UAE and albumin:creatinine ratio (ACR) in the 4-h sample. However, the strength of the correlation between ACR and 24-h UAE was weaker than between the 4- and 24-h UAE. This suggests that normalising to creatinine may not provide additional value to the 4-h urine collection. In conclusion, the strong correlation between 4- and 6-h UAE and 24-h UAE indicates that the collection period can be considerably reduced. This refinement could reduce stress and increase welfare of the db/db model and potentially be applied to other DN models.

Laboratory animals need to be monitored to check the status of their health and welfare. Routine checks of laboratory fish are limited to visual observations of physical appearance and behaviour, but for species held in opaque-walled tanks, such checks are compromised by restricted views, poor visibility and provoked behaviour. Here, we report our experience of using in-tank underwater cameras to monitor laboratory populations of salmoniforme, perciforme and cypriniforme fish. A range of cameras and lenses were investigated and trialled. A standard VGA resolution analogue camera with a one-third-inch chip and 3.6-mm lens was selected based on size, picture quality, the proportion of tank in view and cost. A shell for the camera and mounting system were designed to minimise size and cleaning and enable flexible positioning within tanks. Cameras were connected via digital encoders to a server, making video available to the general computer network. Data collected from recordings of rainbow trout (Oncorhynchus mykiss) confirmed provoked behaviour, ie a change in distribution and increase in activity in response to direct viewing and feeding. The networked cameras therefore enable remote viewing of undisturbed behaviour in real time, providing clear, lateral views unaffected by water surface effects, and facilitate increased frequency of checking. Case studies illustrate how camera monitoring can aid detection of abnormalities in behaviour (eg lack of feeding, posture, swimming) and appearance (eg clinical signs, such as lesions), enabling earlier interventions. Furthermore, recordings provide a resource for reference and retrospective analysis, and evidence to support severity classification and identify humane end-points.