Journal articles on the topic 'Home-cage testing'

AbstractSingle housing is believed to be chronically stressful and to have a negative impact on welfare and cognition in rats (Rattus norvegicus). However, single housing does not consistently evoke stress-like responses nor does it consistently impair cognitive performance. In an experiment in which all cages were separated by an opaque barrier, single- and pair-housed pigmented (dark-eyed) rats performed equally in a cognitive test and displayed similar levels of anxiety during testing. Additionally, bar biting in the home cage did not differ between the two groups. Stress levels both during cognitive testing and in the home cage were higher than those we have previously reported when rats were housed without opaque barriers between the cages. We conclude that visual interactions between rats in different cages may be of sufficient significance that single housing in a cage with a view to neighbouring rats and to the rest of the laboratory holding room may be preferable to pair housing in a cage without this view.

The cognitive bias test is used to measure the emotional state of animals with regard to future expectations. Thus, the test offers a unique possibility to assess animal welfare with regard to housing and testing conditions of laboratory animals. So far, however, performing such a test is time-consuming and requires the presence of an experimenter. Therefore, we developed an automated and home-cage based cognitive bias test based on the IntelliCage system. We present several developmental steps to improve the experimental design leading to a successful measurement of cognitive bias in group-housed female C57BL/6J mice. The automated and home-cage based test design allows to obtain individual data from group-housed mice, to test the mice in their familiar environment, and during their active phase. By connecting the test-cage to the home-cage via a gating system, the mice participated in the test on a self-chosen schedule, indicating high motivation to actively participate in the experiment. We propose that this should have a positive effect on the animals themselves as well as on the data. Unexpectedly, the mice showed an optimistic cognitive bias after enrichment was removed and additional restraining. An optimistic expectation of the future as a consequence of worsening environmental conditions, however, can also be interpreted as an active coping strategy in which a potential profit is sought to be maximized through a higher willingness to take risks.

Nervous system impairment is prominent among signs of chemical toxicity in humans and animals, yet evaluation of behavioral and neurologic responses is seldom included in premarket screening. The sensitivity and validity of automatically recorded rodent locomotor activity, whether inside or outside of the home cage, justifies its inclusion in first-tier testing. Home cage behaviors are studied in the toxicologic laboratory using quantitative techniques from behavioral neuroscience. A practical, noninvasive, automated system was developed and validated at New York University, in accord with Federal guidelines for testing neurotoxicity. Effects of neurotoxicants on motor activity, eating, drinking, and the daily cycle of rest-activity indicate sensitivity to a variety of chemicals as well as new avenues to the understanding of mechanisms of toxicity. The rat's pattern of nocturnal activity is particularly sensitive to neurotoxicants and thus deserves additional attention. The coefficient of variability of various end points did not correlate with sensitivity to toxicants. This underscores the need for behavioral data to supplement theoretical considerations in test selection. The system's advantages are economy, high data capacity, humaneness, accessible and well-known end points, widely available equipment, automation, and the potential for direct comparisons of several different animal species.

In response to present and anticipated regulatory needs for routine screening of pharmaceuticals for assessment of their neurotoxic potential, a primary tier screen for rodents, consisting of a functional observational battery (FOB) and an automated test of motor activity, has been developed at Searle. Additionally, an FOB for dogs currently is being developed. The rodent FOB assess such functions as home cage and open field activity, stimulus reactivity, and neuromuscular function. The dog FOB emphasizes evaluation of gait, postural reactions, and reflex function. The strategy taken has been to incorporate the primary rodent tier screen into repeated dose preclinical studies. Positive findings would trigger a secondary tier of testing, which would involve the use of more complex and integrated tests of neurobehavioral function. The methodologies for both rodent and nonrodent primary tier screens, validation study results, and the scientific criteria that would trigger secondary tier testing are discussed.

The aim of this study was to develop and refine a heterologous mouse model of endometriosis-associated pain in which non-evoked responses, more relevant to the patient experience, were evaluated. Immunodeficient female mice (N = 24) were each implanted with four endometriotic human lesions (N = 12) or control tissue fat (N = 12) on the abdominal wall using tissue glue. Evoked pain responses were measured biweekly using von Frey filaments. Non-evoked responses were recorded weekly for 8 weeks using a home cage analysis (HCA). Endpoints were distance traveled, social proximity, time spent in the center vs. outer areas of the cage, drinking, and climbing. Significant differences between groups for von Frey response, climbing, and drinking were detected on days 14, 21, and 35 post implanting surgery, respectively, and sustained for the duration of the experiment. In conclusion, a heterologous mouse model of endometriosis-associated evoked a non-evoked pain was developed to improve the relevance of preclinical models to patient experience as a platform for drug testing.

Abstract Aims This study aimed to gain methodological knowledge about quantification of mechanical nociceptive thresholds in pigs stimulated on the hind legs. Methods Thirty-two castrated male pigs were used, weighing 50–60kg, 16 weeks of age and housed in standard pens (4.40m×4.40m) with ad libitum access to water and feed. Mechanical nociceptive testing (4 stimulations/pig) was done by an electronic von Frey anesthesiometer (IITC Life Science Inc., CA) with a rigid, hollow plastic tip (cut-off: 1000 gf) and an area of 0.3 mm2. Stimulations targeted caudal aspects of the metatarsus while the pigs were restrained in a cage measuring 35cm×40cm (length×width). During the last 3d before nociceptive testing, half of the pigs were habituated to the experimental set-up twice daily. Results Across the three days, the habituation procedure led to a reduction in time required to approach and release the pigs from the home pen (P<0.05) and to increased acceptance of the confinement (P<0.01). The habituated animals had a lower mechanical nociceptive threshold (median: 495gf (302–675) vs 745gf (479–1000); P<0.05), and a decreased occurrence of censored observations (0 vs 25%; P<0.05) compared to animals tested without habituation. Conclusions The present experiment focussed on methodological aspects of hand-held tests of mechanical nociceptive thresholds in pigs. The results show that the pigs responded to a 3-day habituation regime by increased ease of handling and tolerance of being confined in a test cage. In addition, habituation to the test environment and handling led to reduced mechanical nociceptive thresholds as well as a lowered proportion of censored observations. The current results reiterate the value of habituation in research involving animal behaviour. Further characterization of the methodology is needed to allow its application in the evaluation of clinical conditions in pigs.

Rodent tests of function have advanced our understanding of movement, largely through the human training and testing and manual assessment. Tools such as reaching and grasping of a food pellet have been widely adopted because they are effective and simple to use. However, these tools are time-consuming, subjective, and often qualitative. Automation of training, testing, and assessment has the potential to increase efficiency while ensuring tasks are objective and quantitative. We detail new methods for automating rodent forelimb tests, including the use of pellet dispensers, sensors, computer vision, and home cage systems. We argue that limitations in existing forelimb tasks are driving the innovations in automated systems. We further argue that automated tasks partially address these limitations, and we outline necessary precautions and remaining challenges when adopting these types of tasks. Finally, we suggest attributes of future automated rodent assessment tools that can enable widespread adoption and help us better understand forelimb function in health and disease.

With the growing awareness of the neurological effects of many environmental chemicals, there is considerable emphasis being placed on the detection of neurotoxic potential at the screening, or first-tier, level of testing. We have developed a neurobehavioral screening battery consisting of a functional observational battery (FOB) and an automated measure of motor activity which can be incorporated into neuropathological studies as well as general toxicity assessments of new and existing chemicals in rats. This approach has been recommended by several expert national and international panels, and recently has been recommended by the U.S. Environmental Protection Agency for testing pesticides and industrial chemicals. The FOB consists of a series of home-cage, open-field, and interactive assessments of the rat's behavior and neurological functions. Motor activity provides an objective apical measure of the rat's neurobehavioral function. In order to assess validity, sensitivity, and specificity, we have utilized these test methods to determine dose-response and time-course characteristics of a wide variety of chemicals including pesticides, metals, solvents, and industrial compounds. We currently are developing approaches to provide profiles of effect for different types of neurotoxicants, which can then be used to make decisions concerning neurotoxic potential and provide direction for more in-depth, second-tier testing.

Behavioral evaluations are emerging as a key component in neurotoxicity testing. A neurobehavioral screening battery that is most often used for hazard identification consists of a functional observational battery (FOB) and motor activity. The FOB was developed as an improvement over routine cageside observations: major differences include the specification of observations, operational definitions of the behaviors to be observed, grading the severity of effect, and location of observations (open arena vs. home cage). A typical FOB protocol consists of approximately 20 to 30 end points, allowing for the detection and description of a range of neurobehavioral changes. We have focused on assessing critical characteristics (i.e., sensitivity, specificity, validity, and reproducibility) of these tests. Concerns have been raised as to the validity and reliability of observational methods for neurotoxicity screening. These tests were therefore the focus of an international collaborative study, sponsored by the International Programme on Chemical Safety (IPCS). The purpose of the IPCS study was to assess the general utility and reliability of neurobehavioral screening procedures in a diversity of testing situations. The test protocol was essentially that described in current United States Environmental Protection Agency (US EPA) guidelines. The resultant data indicated that all participants could detect and characterize the effects of known neurotoxicants, and the divergent profiles of effect obtained for different chemicals demonstrated the specificity of the methods. One conclusion, therefore, was that behavior can be a useful tool for assessing neurotoxicity. In addition, the study also provided a wealth of control data, examination of which provides evidence for the consistency and baseline values of behavioral measures. Reliability estimates were statistically established, providing a measure of the usefulness of individual end points for detecting chemical-induced toxicity. Another conclusion of the IPCS collaborative study was that the application of these methods requires careful attention to details of experimental design, observer training, and experience; these capabilities should be adequately documented by the use of proficiency studies. Recently, modifications of the standard FOB have been introduced, including expanded clinical observations (ECO) for standard toxicity studies. Although there are end points common between the FOB and ECO, it is not known if the latter is sufficient for neurotoxicity screening. Appropriate validation studies have not been conducted on these expanded observations. With the increasing dependency on these methods for neurotoxicity testing, and the number of studies that will be incorporating these methods, issues of test validity as well as data interpretation are becoming more critical.

A functional observational battery (FOB) is recommended as the first-tier neurotoxicity screening in the preclinical safety pharmacology testing guidelines. Minipigs have increasingly been used in regulatory toxicology studies; however, no current FOB protocol is available for neurotoxicity testing in these species. Hence, a minipig FOB instrument was developed. A complete crossover study with Sinclair minipigs was performed to evaluate physiologic, neurologic, and behavioral effects of amphetamine, ketamine, and diazepam. The treated minipigs were first observed in their home cage, were video-recorded for 10 minutes in an open field, and then went through a complete neurologic examination. Both ketamine and diazepam were shown to reduce the freezing and behavior shifts of treated minipigs, while increasing their exploratory behaviors. Both drugs also caused muscular and gait impairment. The effects of ketamine and diazepam were consistent with their roles as central nervous system (CNS) suppressants. Unique effects were also observed with ketamine and diazepam treatments, which may reflect their unique mechanisms of action. Consistent with its role as a CNS stimulant, amphetamine caused the treated minipigs to be hyperactive and to display increased freezing and behavior shifts and reduced exploring activities. These effects of amphetamine were opposite to those observed with ketamine and diazepam. Amphetamine also increased locomotion in the treated minipigs. The present effects of amphetamine, ketamine, and diazepam are in agreement with observations by others. In conclusion, the minipig is a suitable species for FOB evaluation of pharmaceuticals in preclinical safety pharmacology testing.

Abstract Background Pavlovian stimuli predictive of appetitive outcomes can exert a powerful influence on the selection and initiation of action, a phenomenon termed outcome-selective Pavlovian-instrumental transfer (sPIT). Rodent studies suggest that sPIT is insensitive to motivational downshift induced by outcome devaluation, an effect that is, however, relatively underexplored. Methods Here we examined in detail the effects of distinct shifts in motivation from hunger to a state of relative satiety on sPIT in rats. Results A motivational downshift by outcome-specific devaluation immediately prior to testing markedly reduced overall lever responding and magazine entries but left intact the sPIT effect. A motivational downshift prior testing by (1) giving ad libitum rather than restricted access to maintenance diet in the home cage for 24 hours or by (2) a systemic blockade of hormone secretagogue receptor subtype 1A receptors to inhibit orexigenic actions of ghrelin both reduced overall lever responding and magazine entries. Moreover, these latter motivational downshifts reduced the sPIT effect; however, the sizes of the sPIT effects were still large. Conclusions Collectively, our rodent findings indicate that major effects of various motivational downshifts are overall inhibition of lever pressing and magazine approach, possibly reflecting reduced general motivation. The observed effects of motivational downshifts on sPIT have implications with regard to the role of general motivating effects in sPIT and to the contribution of Pavlovian-instrumental interactions to excessive food seeking as well as obesity in humans.

In the adoption of behavior as a critical end point in safety pharmacology and neurotoxicity screening, federal regulatory agencies have shifted the predominating scientific perspective from pharmacology back to the experimental analysis of behavior (psychology). Nowhere is this more evident than in tier I safety assessment of the central nervous system (CNS). The CNS and peripheral nervous system have multiple behavioral units of general activity. A complete picture of the motor control neural pathways cannot be measured by any one single approach. The CNS safety protocols under International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use S7A are required to be conducted in accordance with Good Laboratory Practices by trained technical staff. The CNS safety assessments necessitate the inclusion of a thorough and detailed behavioral analysis of home cage activity, the response to handling, and transportation to and observations within an open-field apparatus with ancillary measures of basal muscle tone, muscle strength, and tremor in a functional observation battery, as well as quantitative measurements of 3-dimensional activity in an automated photobeam arena. Cost-cutting initiatives or a radical application of the “reduce use” principle of the 3 Rs only jeopardize the spirit, intent, and predictive validity of tier I safety testing assays dictated by current drug safety guidelines.

Abstract Increasing evidence suggests that stimulus temperature modifies taste signaling. However, understanding how temperature modifies taste-driven behavior is difficult to separate as we must first understand how temperature alone modifies behavior. Previous work has suggested that cold water is more rewarding and “satiating” than warm water, and water above orolingual temperature is avoided in brief-access testing. We explored the strength of cold water preference and warm water avoidance by asking: (1) if cold temperature alone was sufficient to condition a flavor preference and (2) if avoidance of warm stimuli is driven by novelty. We addressed these questions using custom-designed equipment that allows us to monitor and maintain solution temperatures. We conducted two-bottle preference tests, after pairing Kool-Aid flavors with 10 or 40 °C. Rats preferred the flavor paired with cold temperature, both while it was cold and for 1 day while solutions were presented at 22 °C. We then examined the role of novelty in avoidance of 40 °C. Rats were maintained on 10, 22, or 40 °C water in their home cage to increase familiarity with the temperatures. Rats were then subject to a series of brief-access taste tests to water or sucrose at 10 to 40 °C. Rats that had 40 °C experience licked more to 40 °C water, but not sucrose, during brief-access testing. In a series of two-bottle preference tests, rats maintained on 40 °C water had a decreased preference for 10 °C water when paired opposite 40 °C water. Together, these data contribute to our understanding of orosensory-driven behavior with water at different temperatures.

C57BL/6NTac (Bl6) and 129S6/SvEvTac (129Sv) mice display different coping strategies in stressful conditions. Our aim was to evaluate biomarkers related to different adaptation strategies in the brain of male 129Sv and Bl6 mice. We focused on signaling pathways related to the dopamine (DA) system, N-methyl-D-aspartate (NMDA) receptor and epidermal growth factor (EGF) family, shown as the key players in behavioral adaptation. Mice from Bl6 and 129Sv lines were divided into either home cage controls (HCC group) or exposed to repeated motility testing and treated with saline for 11 days (RMT group). Distinct stress responses were reflected in severe body weight loss in 129Sv and the increased exploratory behavior in Bl6 mice. Besides that, amphetamine caused significantly stronger motor stimulation in Bl6. Together with the results from gene expression (particularly Maob), this study supports higher baseline activity of DA system in Bl6. Interestingly, the adaptation is reflected with opposite changes of DA markers in dorsal and ventral striatum. In forebrain, stress increased the gene expressions of Egf-Erbb1 and Nrg1/Nrg2-Erbb4 pathways more clearly in 129Sv, whereas the corresponding proteins were significantly elevated in Bl6. We suggest that not only inhibited activity of the DA system, but also reduced activity of EGF family and NMDA receptor signaling underlies higher susceptibility to stress in 129Sv. Altogether, this study underlines the better suitability of 129Sv for modelling neuropsychiatric disorders than Bl6.

Object Heme toxicity may contribute to the pathogenesis of intracerebral hemorrhage (ICH). The primary defense against extracellular heme is provided by hemopexin, a serum and neuronal glycoprotein that binds it with very high affinity and mitigates its prooxidant effect. In the present study, the authors tested the hypothesis that hemopexin knockout mice would sustain more injury after experimental ICH than their wild-type counterparts. Methods: Striatal ICH was induced by the stereotactic injection of bacterial collagenase or autologous blood. Three days later, striatal protein oxidation was assessed via carbonyl assay. Cell viability was quantified at 8–9 days by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Behavioral deficits were detected with high-resolution digital analysis of 6-hour home cage video recordings and standard testing. Results: Perihematomal protein oxidation was increased in wild-type collagenase-injected striata by approximately 2.1-fold, as compared with contralateral striata; protein carbonyls were increased 3-fold in knockout mice. Striatal cell viability was reduced by collagenase injection in wild-type mice to 52.9 ± 6.5% of that in the contralateral striata, and to 31.1 ± 3.7% of that in the contralateral striata in knockout mice; similar results were obtained after blood injection. Digital analysis of 6-hour video recordings demonstrated an activity deficit in both models that was significantly exacerbated at 8 days in knockout mice. Striatal heme content 9 days after blood injection was increased approximately 2.7-fold in knockouts as compared with wild-type mice. Conclusions: These results suggest that hemopexin has a protective effect against hemorrhagic CNS injuries. Hemopexin deficiency, which is often associated with sickle cell disease, may worsen outcome after ICH.

Model organisms mimicking the pathogenesis of human diseases are useful for identifying pathogenic mechanisms and testing therapeutic efficacy of compounds targeting them. Models of Alzheimer’s disease (AD) and related dementias (ADRD) aim to reproduce the brain pathology associated with these neurodegenerative disorders. Transgenic models, which involve random insertion of disease-causing genes under the control of artificial promoters, are efficient means of doing so. There are confounding factors associated with transgenic approaches, however, including target gene overexpression, dysregulation of endogenous gene expression at transgenes’ integration sites, and limitations in mimicking loss-of-function mechanisms. Furthermore, the choice of species is important, and there are anatomical, physiological, and cognitive reasons for favoring the rat over the mouse, which has been the standard for models of neurodegeneration and dementia. We report an initial assessment of the spatial learning, reversal, and sequencing task capabilities of knock-in (KI) Long-Evans rats with humanizing mutations in the Aβ-coding region of App, which encodes amyloid precursor protein (Apph/h rats), using the IntelliCage, an automated operant social home cage system, at 6–8 weeks of age, then again at 4–5 months of age. These rats were previously generated as control organisms for studies on neurodegeneration involving other knock-in rat models from our lab. Apph/h rats of either sex can acquire place learning and reversal tasks. They can also acquire a diagonal sequencing task by 6–8 weeks of age, but not a more advanced serial reversal task involving alternating diagonals, even by 4–5 months of age. Thus, longitudinal behavioral analysis with the IntelliCage system can be useful to determine, in follow-up studies, whether KI rat models of Familial AD (FAD), sporadic late onset AD (LOAD), and of ADRD develop aging-dependent learning and memory deficits.

Abstract Survivors of critical illness often report traumatic memories of their illness period, and these memories are thought to contribute to development of neuropsychiatric disorders, such as PTSD. Many patients are treated with high doses of glucocorticoids for their vasoactive and anti-inflammatory properties, and glucocorticoids have also been shown to prevent the development of PTSD after trauma. Due to their activity in the hippocampus and amygdala, the putative protective effect of glucocorticoids may occur via memory formation during illness. To examine the effect of glucocorticoids on memory formation during acute infectious illness, male and female C57BL/6 mice (N=80, 40 male/40 female) underwent cecal ligation and puncture and were treated with either corticosterone (16 mg/kg) or vehicle in the early afternoon daily for five days beginning on the day of surgery. All mice were habituated to a neutral object in their home cage for five days and underwent one 30-minute footshock/no shock training session during the illness period. After physiologic recovery (2 weeks), the mice underwent behavioral testing including open field exploration, object recognition testing in which they were presented with both the familiar (habituated) object and a novel object, and testing in the shock context. The results showed that drug treatment had no effect on behavior in the open field, including time spent in the center (VEH: 20.19±10.81 vs CORT: 22.32±12.87 sec; P=0.476). Drug treatment increased overall object exploration (12.28±10.79 vs 19.17±15.88 sec; P=0.049). Corticosterone-treated mice showed a preference for the familiar object (60.9±23.0% of total exploration time with familiar object; P=0.015), while vehicle-treated mice did not (54.1±23.3%; P=0.378). The increase in overall object exploration seen in corticosterone-treated mice could be accounted for by an increase in exploration of the familiar object. History of footshock increased freezing in the training context (3.96±2.54% vs 36.08±15.42%; P&lt;0.0001) and corticosterone treatment had no effect (18.06±17.65% vs 22.16±21.19%; P=0.557). In conclusion, administration of corticosterone during infectious illness facilitated memory of a neutral object from the illness period, and recovered mice exhibited a preference for this object over a novel one. Corticosterone treatment had no impact on fear memory formed during illness. This is consistent with human literature suggesting that hydrocortisone decreases PTSD symptoms without impacting traumatic memories. These findings suggest that glucocorticoids selectively enhance the formation, consolidation, and/or recall of neutral but not fear memories during illness, which may rely on hippocampal circuitry. We further suggest that accurate memories of the illness period may influence patients’ perception of this experience and alter their risk for psychiatric sequelae.

Background Animal models used to study pathologies requiring rehabilitation therapy, such as cardiovascular and neurologic disorders or oncologic disease, must be as refined and translationally relevant as possible. Sometimes, however, experimental procedures such as those involving restraint may generate undesired effects which may act as a source of bias. However, the extent to which potentially confounding effects derive from such routine procedures is currently unknown. Our study was therefore aimed at exploring possible undesirable effects of acute restraint stress, whereby animals were exposed to a brightly lit enclosed chamber (R&L) similar to those that are commonly used for substance injection. We hypothesised that this would induce a range of unwanted physiological alterations [such as neuroinflammatory response and changes in body weight and in brown adipose tissue (BAT)] and behavioural modification, and that these might be mitigated via the use of non-aversive handling methods: Tunnel Handling (NAH-T) and Mechanoceptive Handling (NAH-M)) as compared to standard Tail Handling (TH). Methods Two indicators of physiological alterations and three potentially stress sensitive behavioural parameters were assessed. Physiological alterations were recorded via body weight changes and assessing the temperature of Brown Adipose Tissue (BAT) using infra-red thermography (IRT), and at the end of the experiment we determined the concentration of cytokines CXCL12 and CCL2 in bone marrow (BM) and activated microglia in the brain. Nest complexity scoring, automated home-cage behaviour analysis (HCS) and Elevated Plus Maze testing (EPM) were used to detect any behavioural alterations. Recordings were made before and after a 15-minute period of R&L in groups of mice handled via TH, NAH-T or NAH-M. Results BAT temperature significantly decreased in all handling groups following R&L regardless of handling method. There was a difference, at the limit of significance (p = 0.06), in CXCL12 BM content among groups. CXCL12 content in BM of NAH-T animals was similar to that found in Sentinels, the less stressed group of animals. After R&L, mice undergoing NAH-T and NAH-M showed improved body-weight maintenance compared to those exposed to TH. Mice handled via NAH-M spent a significantly longer time on the open arms of the EPM. The HCS results showed that in all mice, regardless of handling method, R&L resulted in a significant reduction in walking and rearing, but not in total distance travelled. All mice also groomed more. No difference among the groups was found in Nest Score, in CCL2 BM content or in brain activated microglia. Conclusions Stress induced by a common restraint procedure caused metabolic and behavioural changes that might increase the risk of unexpected bias. In particular, the significant decrease in BAT temperature could affect the important metabolic pathways controlled by this tissue. R&L lowered the normal frequency of walking and rearing, increased grooming and probably carried a risk of low-grade neuro-inflammation. Some of the observed alterations can be mitigated by Non-aversive handlings.

Ever-increasing demands for energy, particularly being environmentally friendly have promoted the transition from fossil fuels to renewable energy.1 Lithium-ion batteries (LIBs), arguably the most well-studied energy storage system, have dominated the energy market since their advent in the 1990s.2 However, challenging issues regarding safety, supply of lithium, and high price of lithium resources limit the further advancement of LIBs for large-scale energy storage applications.3 Therefore, attention is being concentrated on an alternative electrochemical energy storage device that features high safety, low cost, and long cycle life. Rechargeable aqueous zinc-ion batteries (ZIBs) is considered one of the most promising alternative energy storage systems due to the high theoretical energy and power densities where the multiple electrons (Zn2+) . In addition, aqueous ZIBs are safer due to non-flammable electrolyte (e.g., typically aqueous solution) and can be manufactured since they can be assembled in ambient air conditions. 4 As an essential component in aqueous Zn-based batteries, the Zn metal anode generally suffers from the growth of dendrites, which would affect battery performance in several ways. Second, the led by the loose structure of Zn dendrite may reduce the coulombic efficiency and shorten the battery lifespan.5 Several approaches were suggested to improve the electrochemical stability of ZIBs, such as implementing an interfacial buffer layer that separates the active Zn from the bulk electrolyte.6 However, the and thick thickness of the conventional Zn metal foils remain a critical challenge in this field, which may diminish the energy density of the battery drastically. According to a theretical calculation, the thickness of a Zn metal anode with an areal capacity of 1 mAh cm-2 is about 1.7 μm. However, existing extrusion-based fabrication technologies are not capable of downscaling the thickness Zn metal foils below 20 μm. Herein, we demonstrate a thickness controllable coating approach to fabricate an ultrathin Zn metal anode as well as a thin dielectric oxide separator. First, a 1.7 μm Zn layer was uniformly thermally evaporated onto a Cu foil. Then, Al2O3 , the separator was deposited through sputtering on the Zn layer to a thickness of 10 nm. The inert and high hardness Al2O3 layer is expected to lower the polarization and restrain the growth of Zn dendrites. Atomic force microscopy was employed to evaluate the roughness of the surface of the deposited Zn and Al2O3/Zn anode structures. Long-term cycling stability was gauged under the symmetrical cells at 0.5 mA cm-2 for 1 mAh cm-2. Then the fabricated Zn anode was paired with MnO2 as a full cell for further electrochemical performance testing. To investigate the evolution of the interface between the Zn anode and the electrolyte, a home-developed in-situ optical observation battery cage was employed to record and compare the process of Zn deposition on the anodes of the Al2O3/Zn (demonstrated in this study) and the procured thick Zn anode. The surface morphology of the two Zn anodes after circulation was characterized and compared through scanning electron microscopy. The tunable ultrathin Zn metal anode with enhanced anode stability provides a pathway for future high-energy-density Zn-ion batteries. Obama, B., The irreversible momentum of clean energy. Science 2017, 355 (6321), 126-129. Goodenough, J. B.; Park, K. S., The Li-ion rechargeable battery: a perspective. J Am Chem Soc 2013, 135 (4), 1167-76. Li, C.; Xie, X.; Liang, S.; Zhou, J., Issues and Future Perspective on Zinc Metal Anode for Rechargeable Aqueous Zinc‐ion Batteries. Energy & Environmental Materials 2020, 3 (2), 146-159. Jia, H.; Wang, Z.; Tawiah, B.; Wang, Y.; Chan, C.-Y.; Fei, B.; Pan, F., Recent advances in zinc anodes for high-performance aqueous Zn-ion batteries. Nano Energy 2020, 70. Yang, J.; Yin, B.; Sun, Y.; Pan, H.; Sun, W.; Jia, B.; Zhang, S.; Ma, T., Zinc Anode for Mild Aqueous Zinc-Ion Batteries: Challenges, Strategies, and Perspectives. Nanomicro Lett 2022, 14 (1), 42. Yang, Q.; Li, Q.; Liu, Z.; Wang, D.; Guo, Y.; Li, X.; Tang, Y.; Li, H.; Dong, B.; Zhi, C., Dendrites in Zn-Based Batteries. Adv Mater 2020, 32 (48), e2001854. Acknowledgment This work was partially supported by the U.S. National Science Foundation (NSF) Award No. ECCS-1931088. S.L. and H.W.S. acknowledge the support from the Improvement of Measurement Standards and Technology for Mechanical Metrology (Grant No. 22011044) by KRISS. Figure 1

Abstract While metabolic disorders such as obesity and diabetes are costly and deadly to the current population, they are also extremely detrimental to the next generation. Much of the current literature focuses on the negative impact of poor maternal choices on offspring disease, while there is little work examining maternal behaviors that may improve offspring health. Research has shown that voluntary maternal exercise in mouse models improves metabolic function in offspring. In this study, we hypothesized that controlled maternal exercise in a mouse model will effect positive change on offspring obesity and glucose homeostasis. Female mice were separated into three groups: home cage, sedentary, and exercise. The sedentary home cage group was not removed from the home cage, while the sedentary wheel group was removed from the cage and placed in an immobile wheel apparatus. The exercise group was removed from the home cage and run on the same wheel apparatus but with the motor activated at 5–10 m/min for 1 h/d prior to and during pregnancy. Offspring were subjected to oral glucose tolerance testing and body composition analysis. There was no significant difference in offspring glucose tolerance or body composition as a consequence of the maternal exercise intervention compared to the sedentary wheel group. There were no marked negative consequences of the maternal controlled exercise intervention. Further research should clarify the potential advantages of the controlled exercise model and improve experimental techniques to facilitate translation of this research to human applications.

The reproducibility crisis (or replication crisis) in biomedical research is a particularly existential and under-addressed issue in the field of behavioral neuroscience, where, in spite of efforts to standardize testing and assay protocols, several known and unknown sources of confounding environmental factors add to variance. Human interference is a major contributor to variability both within and across laboratories, as well as novelty-induced anxiety. Attempts to reduce human interference and to measure more "natural" behaviors in subjects has led to the development of automated home-cage monitoring systems. These systems enable prolonged and longitudinal recordings, and provide large continuous measures of spontaneous behavior that can be analyzed across multiple time scales. In this review, a diverse team of neuroscientists and product developers share their experiences using such an automated monitoring system that combines Noldus PhenoTyper® home-cages and the video-based tracking software, EthoVision® XT, to extract digital biomarkers of motor, emotional, social and cognitive behavior. After presenting our working definition of a “home-cage”, we compare home-cage testing with more conventional out-of-cage tests (e.g., the open field) and outline the various advantages of the former, including opportunities for within-subject analyses and assessments of circadian and ultradian activity. Next, we address technical issues pertaining to the acquisition of behavioral data, such as the fine-tuning of the tracking software and the potential for integration with biotelemetry and optogenetics. Finally, we provide guidance on which behavioral measures to emphasize, how to filter, segment, and analyze behavior, and how to use analysis scripts. We summarize how the PhenoTyper has applications to study neuropharmacology as well as animal models of neurodegenerative and neuropsychiatric illness. Looking forward, we examine current challenges and the impact of new developments. Examples include the automated recognition of specific behaviors, unambiguous tracking of individuals in a social context, the development of more animal-centered measures of behavior and ways of dealing with large datasets. Together, we advocate that by embracing standardized home-cage monitoring platforms like the PhenoTyper, we are poised to directly assess issues pertaining to reproducibility, and more importantly, measure features of rodent behavior under more ethologically relevant scenarios.

In animal experimentation, welfare and severity assessments of all procedures applied to animals are necessary to meet legal and ethical requirements, as well as public interests. So far, the methods suggested for this purpose are time consuming and personnel intensive. Also, evidence-based biostatistical methods for this purpose are still rare. We here tested whether the classification of heart rate (HR) and activity (Act) data monitored by telemetry in the home cage by unsupervised k-means-based class-labeling and subsequent Support Vector Machine (SVM) analysis allows severity assessment and grading of experimental procedures of different domains, including surgery, injection, behavioral testing, and routine handling for maintenance. Telemetric devices were subcutaneously implanted in young adult male Crl:CD(SD) and BDIX/UImHanZtm rats. After recovery, rats were randomly subjected to different experimental procedures, i.e., handling and cage change as routine maintenance, Rat Grimace Scale, burrowing, and social interaction for welfare assessment, as well as repeated subcutaneous injections. Thereafter, rats were either intracranially implanted with electrodes or injected with tumor cells. Directly after each procedure, HR and Act were monitored by telemetry in the home cage for 4 h. Application of k-means and SVM algorithms on the obtained data sets from baseline (as no stress), cage change (exploratory Act), and intracranial surgery (as burden) measurements computed three classes described as low HR/low Act, high HR/high Act, and high HR/low Act, respectively. Validation of the SVM model by entering data from all procedures confirmed the allocation to the high HR/low Act class (burden) after surgery, which lasted longer after subcutaneous transmitter implantation than after intracranial surgery. The majority of data points from repeated injections, behavioral testing, and maintenance handling were allocated to the low HR/low Act and high HR/high Act classes. Overall, the SVM model based on HR and Act data monitored in home cage after procedures may be useful for the classification and grading of experimental procedures of different domains.

Laboratory mice in standard laboratory cages, besides horizontal and vertical locomotor activity, spontaneously display cage-bar related activities such as cage-grid climbing. Although, grid-climbing activity is one of the major components of spontaneous home-cage behavior of mice, its exact role is not fully understood. This study aimed to describe the sex-differences in coping with novelty and in spontaneous behavior of laboratory mice concerning the cage-climbing activity in an observer-independent open field test. Adult mice of both sexes (C57Bl/6NTac) underwent behavioral testing in LABORAS system. Female mice travelled significantly longer distance (by 30 %, p<0.05) and showed higher grid-climbing activity (by 50 %, p<0.05) than males. Based on our results, the grid-climbing is a sex-dependent activity of mice, however, its exact role remains to be elucidated.

Electrophysiological studies with behaving non-human primates (NHP) often require the separation of animals from their social group as well as partial movement restraint to perform well controlled experiments. When the research goal per se does not mandate constraining the animals’ movements there are often still experimental needs imposed by tethered data acquisition. Recent technological advances meanwhile allow wireless neurophysiological recordings at high band-width in limited-size enclosures. Here, we demonstrate wireless neural recordings at single unit resolution from unrestrained Rhesus macaques while they performed self-paced, structured visuomotor tasks on our custom-built, stand-alone touchscreen system (XBI) in their home environment. We were able to successfully characterize neural tuning to task parameters, such as visuo-spatial selectivity during movement planning and execution, as expected from existing findings obtained via setup-based neurophysiology recordings. We conclude that when movement restraint and/or a highly controlled, insulated environment are not necessary for scientific reasons, cage-based wireless neural recordings are a viable option. We propose an approach that allows the animals to engage in a self-paced manner with our XBI device, both for fully automatized training and cognitive testing, as well as neural data acquisition in their familiar environment, maintaining auditory and sometimes visual contact with their conspecifics.Significance statementCage-based cognitive systems have previously been shown to be highly useful in cognitive assessment of non-human primates. These systems allow animals to engage with the task/system in an unrestrained and self-paced manner. We expanded the capabilities of our own cage-based testing device by combining cognitive testing with wireless neural recordings in the animals’ home environment, in an upscalable approach. When neither movement constraints nor specialized equipment are scientifically necessary, our approach allows for the combination of cognitive testing with intracranial electrophysiology without removing the animal from its home environment, potentially improving animal well-being.

Automated mouse phenotyping through the high-throughput analysis of home cage behavior has brought hope of a more effective and efficient method for testing rodent models of diseases. Advanced video analysis software is able to derive behavioral sequence data sets from multiple-day recordings. However, no dedicated mechanisms exist for sharing or analyzing these types of data. In this article, we present a free, open-source software actionable through a web browser (an R Shiny application), which performs an analysis of home cage behavioral sequence data, which is designed to spot differences in circadian activity while preventing p-hacking. The software aligns time-series data to the light/dark cycle, and then uses different time windows to produce up to 162 behavior variables per animal. A principal component analysis strategy detected differences between groups. The behavior activity is represented graphically for further explorative analysis. A machine-learning approach was implemented, but it proved ineffective at separating the experimental groups. The software requires spreadsheets that provide information about the experiment (i.e., metadata), thus promoting a data management strategy that leads to FAIR data production. This encourages the publication of some metadata even when the data are kept private. We tested our software by comparing the behavior of female mice in videos recorded twice at 3 and 7 months in a home cage monitoring system. This study demonstrated that combining data management with data analysis leads to a more efficient and effective research process.

A major hurdle preventing effective interventions for patients with mild traumatic brain injury (mTBI) is the lack of known mechanisms for the long-term cognitive impairment that follows mTBI. The closed head impact model of repeated engineered rotational acceleration (rCHIMERA), a non-surgical animal model of repeated mTBI (rmTBI), mimics key features of rmTBI in humans. Using the rCHIMERA in rats, this study was designed to characterize rmTBI-induced behavioral disruption, underlying electrophysiological changes in the medial prefrontal cortex (mPFC), and associated mitochondrial dysfunction. Rats received 6 closed-head impacts over 2 days at 2 Joules of energy. Behavioral testing included automated analysis of behavior in open field and home-cage environments, rotarod test for motor skills, novel object recognition, and fear conditioning. Following rmTBI, rats spent less time grooming and less time in the center of the open field arena. Rats in their home cage had reduced inactivity time 1 week after mTBI and increased exploration time 1 month after injury. Impaired associative fear learning and memory in fear conditioning test, and reduced short-term memory in novel object recognition test were found 4 weeks after rmTBI. Single-unit in vivo recordings showed increased neuronal activity in the mPFC after rmTBI, partially attributable to neuronal disinhibition from reduced inhibitory synaptic transmission, possibly secondary to impaired mitochondrial function. These findings help validate this rat rmTBI model as replicating clinical features, and point to impaired mitochondrial functions after injury as causing imbalanced synaptic transmission and consequent impaired long-term cognitive dysfunction.

Alterations of glutamatergic neurotransmission have been implicated in neurodevelopmental and neuropsychiatric disorders. Mice lacking the GluA1 AMPA receptor subunit, encoded by the Gria1 gene, display multiple phenotypical features associated with glutamatergic dysfunction. While the phenotype of adult GluA1 deficient (Gria1–/–) mice has been studied comprehensively, there are relevant gaps in knowledge about the course and the onset of behavioral alterations in the Gria1 knockout mouse model during post-weaning development. Based on former investigations in young wild-type mice, we exposed female and male adolescent Gria1–/– mice to a behavioral home-cage based testing battery designed for the purpose of severity assessment. Data obtained from mice with a constitutive loss of GluA1 were compared with those from wild-type littermates. We identified several genotype-dependent behavioral alterations in young Gria1–/– mice. While the preference for sweetness was not affected by genotype during adolescence, Gria1–/– mice displayed limited burrowing performance, and reached lower nest complexity scores. Analysis of home-cage based voluntary wheel running performance failed to confirm genotype-dependent differences. In contrast, when exposed to the open field test, Gria1–/– mice showed pronounced hyperlocomotion in early and late adolescence, and female Gria1–/– mice exhibited thigmotaxis when prepubescent. We found increased corticosterone metabolite levels in fecal samples of adolescent Gria1–/– mice with females exhibiting increased adrenocortical activity already in prepubescence. Considering the course of behavioral modifications in early and late adolescence, the results do not support a persistent level of distress associated with GluA1 deficiency in the line. In contrast, the laboratory-specific readouts indicate transient, mild impairments of behavioral patterns relevant to animal welfare, and suggest a mild overall burden of the line.

Water regulation is a procedure that allows animals to consume water volumes equivalent to ad libitum access, but accessis limited to specific time intervals (that is, water is not available outside of the designated access periods). Despite the relatively common use of water regulation in research, the implementation method is rarely detailed, stating only that water was available in the animal’s home cage at specific times. For planned toxicologic assessments, we placed rats (n = 510) and ferrets (n = 16) on water regulation using both automated and manual methods. In testing our systems, we defined “successful implementation” as maintenance of appropriate weight gain and health status. An automated system that controlled water access to an entire rat rack was successful for most rats, but several rats failed to consume enough water even after 2 wk of experience. Manual methods of water regulation were successful in rats by either moving the cage to prevent access to the drinking valve or by placing/removing water bottles. An automated system that controlled water access from water bottles was implemented for ferrets and was maintained for up to 30 wk. Retrospective comparison of body weights to standard growth curves for both species showed that all animals grew normally despite water regulation. Differences in the systems and some species considerations provide insights into the key elements necessary for successful water regulation in rats and ferrets.

A primary goal in pain treatment is restoration of behaviors that are disrupted by pain. Measures of pain interference indicate the degree to which pain interferes with activities in pain patients, and these measures are used to evaluate the effects of analgesic drugs. As a result of the emphasis on the expression and treatment of functional impairment in clinical settings, preclinical pain researchers have attempted to develop procedures for evaluation of pain-related functional impairment in laboratory animals. The goal of the present study was to develop and validate a low cost procedure for the objective evaluation of pain-related depression of home cage behavior in mice. On test days, a 5 × 5 cm Nestlet was weighed prior to being suspended from the wire lid of the home cage of individually housed male and female ICR mice. Over the course of experimental sessions, mice removed pieces of the suspended Nestlet, and began to build a nest with the material they removed. Thus, the weight of the pieces of Nestlet that remained suspended at various time points in the session provided an indicator of the rate of this behavior. The results indicate that Nestlet shredding was stable with repeated testing, and shredding was depressed by intra-peritoneal injection of 0.32% lactic acid. The non-steroidal anti-inflammatory drug ketoprofen blocked 0.32% lactic acid-induced depression of shredding, but did not block depression of shredding by a pharmacological stimulus, the kappa opioid receptor agonist U69,593. The μ-opioid receptor agonist morphine did not block 0.32% lactic acid-induced depression of shredding when tested up to doses that depressed shredding in the absence of lactic acid. When noxious stimulus intensity was reduced by decreasing the lactic acid concentration to 0.18%, morphine was effective at blocking pain-related depression of behavior. In summary, the data from the present study support consideration of the Nestlet shredding procedure for use in studies examining mechanisms, expression, and treatment of pain-related functional impairment.

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder, with a long preclinical and prodromal phase. To enable the study of disease mechanisms, AD has been modeled in many transgenic animal lines and cognitive functioning has been tested using several widely used behavioral tasks. These tasks, however, are not always suited for repeated longitudinal testing and are often associated with acute stress such as animal transfer, handling, novelty, or stress related to the task itself. This makes it challenging to relate cognitive dysfunction in animal models to cognitive decline observed in AD patients. Here, we designed an automated figure-8-maze (F8M) to test mice in a delayed alternation task (DAT) in a longitudinal manner. Mice were rewarded when they entered alternate sides of the maze on subsequent trials. Automation as well as connection of the F8M set-up with a home cage reduces experimenter interference and minimizes acute stress, thus making it suitable for longitudinal testing and facilitating clinical translation. In the present study, we monitored cognitive functioning of 2-month-old APPswe/PSEN1dE9 (APP/PS1) mice over a period of 4 months. The percentage of correct responses in the DAT did not differ between wild-type and transgenic mice from 2 to 6 months of age. However, 6-month-old mice displayed an increase in the number of consecutive incorrect responses. These results demonstrate the feasibility of longitudinal testing using an automated F8M and suggest that APP/PS1 mice are not impaired at delayed spatial alternation until 6 months of age under the current experimental conditions.

AbstractExisting methods for analysis of home cage-based preference tests are either time-consuming, not suitable for group management, expensive, and/or based on proprietary equipment that is not freely available. To correct this, we developed an automated system for group-housed mice based on radio frequency identification: the Mouse Position Surveillance System (MoPSS). The system uses an Arduino microcontroller with compatible components; it is affordable and easy to rebuild for every laboratory because it uses free and open-source software and open-source hardware with the RFID readers as the only proprietary component. The MoPSS was validated using female C57BL/6J mice and manual video comparison. It proved to be accurate even for fast-moving mice (up to 100% accuracy after logical reconstruction), and is already implemented in several studies in our laboratory. Here, we provide the complete construction description as well as the validation data and the results of an example experiment. This tracking system will allow group-based preference testing with individually identified mice to be carried out in a convenient manner. This facilitation of preference tests creates the foundation for better housing conditions from the animals’ perspective.

Electrophysiological and behavioral alterations, including sleep and cognitive impairments, are critical components of age-related decline and neurodegenerative diseases. In preclinical investigation, many refined techniques are employed to probe these phenotypes, but they are often conducted separately. Herein, we provide a protocol for one-time surgical implantation of EMG wires in the nuchal muscle and a skull-surface EEG headcap in mice, capable of 9-to-12-month recording longevity. All data acquisitions are wireless, making them compatible with simultaneous EEG recording coupled to multiple behavioral tasks, as we demonstrate with locomotion/sleep staging during home-cage video assessments, cognitive testing in the Barnes maze, and sleep disruption. Time-course EEG and EMG data can be accurately mapped to the behavioral phenotype and synchronized with neuronal frequencies for movement and the location to target in the Barnes maze. We discuss critical steps for optimizing headcap surgery and alternative approaches, including increasing the number of EEG channels or utilizing depth electrodes with the system. Combining electrophysiological and behavioral measurements in preclinical models of aging and neurodegeneration has great potential for improving mechanistic and therapeutic assessments and determining early markers of brain disorders.

Abstract Background An active lifestyle is associated with improved cognitive functions in aged people and may prevent or slow down the progression of various neurodegenerative diseases including Alzheimer’s disease (AD). To investigate these protective effects, male APPNL-G-F mice were exposed to long-term voluntary exercise. Methods Three-month-old AD mice were housed in a cage supplemented with a running wheel for 9 months for long-term exercise. At the age of 12 months, behavioral tests were completed for all groups. After completing behavioral testing, their brains were assessed for amyloid pathology, microgliosis, and cholinergic cells. Results The results showed that APPNL-G-F mice allowed to voluntarily exercise showed an improvement in cognitive functions. Furthermore, long-term exercise also improved anxiety in APPNL-G-F mice as assessed by measuring thigmotaxis in the Morris water task. We also found reductions in amyloid load and microgliosis, and a preservation of cholinergic cells in the brain of APPNL-G-F mice allowed to exercise in their home cages. These profound reductions in brain pathology associated with AD are likely responsible for the observed improvement of learning and memory functions following extensive and regular exercise. Conclusion These findings suggest the potential of physical exercise to mitigate the cognitive deficits in AD.

Abstract The synaptic homeostasis theory of sleep proposes that low neurotransmitter activity in sleep optimizes memory consolidation. We tested this theory by asking whether increasing acetylcholine levels during early sleep would weaken motor memory consolidation. We trained separate groups of adult mice on the rotarod walking task and the single pellet reaching task, and after training, administered physostigmine, an acetylcholinesterase inhibitor, to increase cholinergic tone in subsequent sleep. Post-sleep testing showed that physostigmine impaired motor skill acquisition of both tasks. Home-cage video monitoring and electrophysiology revealed that physostigmine disrupted sleep structure, delayed non-rapid-eye-movement sleep onset, and reduced slow-wave power in the hippocampus and cortex. Additional experiments showed that: (1) the impaired performance associated with physostigmine was not due to its effects on sleep structure, as 1 h of sleep deprivation after training did not impair rotarod performance, (2) a reduction in cholinergic tone by inactivation of cholinergic neurons during early sleep did not affect rotarod performance, and (3) stimulating or blocking muscarinic and nicotinic acetylcholine receptors did not impair rotarod performance. Taken together, the experiments suggest that the increased slow wave activity and inactivation of both muscarinic and nicotinic receptors during early sleep due to reduced acetylcholine contribute to motor memory consolidation.

Prior studies link anxiety to alterations in serotonergic neurotransmission. Tao and Menard (unpublished data) found that female rats exposed to intermittent physical stress during early adolescence display greater levels of anxiety-like behaviours when tested as adults in social interaction tests. Interestingly, these behavioural changes were associated with changes in serotonin (5-HT) fiber density in the medial prefrontal cortex. Whether these changes in 5-HT fibre density are accompanied by changes in the number of 5-HT-producing cells in the dorsal raphe is not known. To examine this research question, tryptophan-hydroxylase (TPH) immunoreactivity (-IR) was used to label 5-HT cells in the dorsal raphe. Half the animals from the original adolescent stress study were exposed to IPS, and the remaining animals were handled only, serving as the control (CON) group. In adulthood, half of each of those groups were tested in the SI test. The remaining animals were not behaviourally tested, serving as home-cage controls (HCC). A two-factor analysis of variance will be performed with Treatment (IPS vs CON) and Testing (SI vs HCC) as the two factors. It is expected that, regardless of Testing, IPS rats will display higher levels of TPH-IR in the dorsal raphe than no-stress controls. The number of TPH-IR-positive cells will also be correlated with archival behavioural data. It is expected that that higher levels of TPH-IR will be associated with higher levels of anxiety-like behaviour. This study provides insights into the role that early adversity plays in shaping the structure and function of the brain’s serotonergenic system.

In recent years, the utility and efficiency of automated procedures for cognitive assessment in psychology and neuroscience have been demonstrated in non-human primates (NHP). This approach mimics conventional shaping principles of breaking down a final desired behavior into smaller components that can be trained in a staircase manner. When combined with home-cage-based approaches, this could lead to a reduction in human workload, enhancement in data quality, and improvement in animal welfare. However, to our knowledge, there are no reported attempts to develop automated training and testing protocols for long-tailed macaques (Macaca fascicularis), a ubiquitous NHP model in neuroscience and pharmaceutical research. In the current work, we present the results from 6 long-tailed macaques that were trained using an automated unsupervised training (AUT) protocol for introducing the animals to the basics of a two-alternative choice (2 AC) task where they had to discriminate a conspecific vocalization from a pure tone relying on images presented on a touchscreen to report their response. We found that animals (1) consistently engaged with the device across several months; (2) interacted in bouts of high engagement; (3) alternated peacefully to interact with the device; and (4) smoothly ascended from step to step in the visually guided section of the procedure, in line with previous results from other NHPs. However, we also found (5) that animals’ performance remained at chance level as soon as the acoustically guided steps were reached; and (6) that the engagement level decreased significantly with decreasing performance during the transition from visual to acoustic-guided sections. We conclude that with an autonomous approach, it is possible to train long-tailed macaques in their social group using computer vision techniques and without dietary restriction to solve a visually guided discrimination task but not an acoustically guided task. We provide suggestions on what future attempts could take into consideration to instruct acoustically guided discrimination tasks successfully.

Intracerebral hemorrhage (ICH) is a type of hemorrhagic stroke associated with high mortality and morbidity rates. Prolonged and unbalanced inflammatory responses after hemorrhagic stroke may negatively contribute to secondary brain injury. Prostaglandin E2 (PGE 2 ) is an important mediator of inflammation and plays a role in several neurologic disorders. PGE 2 has selective high affinity toward its membrane-bound G-protein-coupled receptors EP1, EP2, EP3, and EP4, which together arbitrate its neuromodulatory effects. We have previously shown in a middle cerebral artery occlusion model of ischemia/reperfusion-induced brain injury and a striatal NMDA-induced excitotoxicity model that mice pretreated intracerebroventricularly with a selective EP3 receptor agonist have larger brain lesions when compared to control mice. In the present study, we investigated whether EP3 receptor deletion (EP3 -/- ) would lead to improved anatomical and functional outcomes in young (2-4mo) and aged (22-24mo) C57BL/6 mice after inducing a hemorrhagic stroke using the collagenase model of ICH (n=8-10 for all groups). Neurobehavioral testing, including video-recording of home cage activity and neurological deficit scoring was performed at 24h, 48h, and 72h post-ICH. At 72h, mice were transcardially perfused and brains isolated for Cresyl Violet staining and lesion volume quantification. In both young and aged mice, smaller brain lesion volumes were seen for the EP3 -/- when compared to age-matched wildtype controls (p<0.05). Further, the aged EP3 -/- displayed significantly better neurologic recovery at 72h, as identified by both behavioral tests (p<0.01). Interestingly, the aged EP3 -/- showed significantly reduced ambulatory and stereotypic activity at 24h post-ictus (p<0.005). No significant behavioral differences were seen in the young cohort. Collectively, these results indicate a dynamic role of the EP3 receptor in mediating anatomical and functional outcomes following ICH. Additional studies are needed in order to clarify the mechanisms involved and identify potential therapeutic paradigms for alleviating the initial PGE 2 -EP3 receptor associated toxicity and promoting its later beneficial effects.

Abstract Background Incentives to promote drinking (“happy hour”) can encourage faster rates of alcohol consumption, especially in women. Sex differences in drinking dynamics may underlie differential health vulnerabilities relating to alcohol in women versus men. Herein, we used operant procedures to model the happy hour effect and gain insight into the alcohol drinking dynamics of male and female rats. Methods Adult male and female Wistar rats underwent operant training to promote voluntary drinking of 10% (w/v) alcohol (8 rats/sex). We tested how drinking patterns changed after manipulating the effort required for alcohol (fixed ratio, FR), as well as the length of time in which rats had access to alcohol (self-administration session length). Rats were tested twice within the 12 h of the dark cycle, first at 2 h (early phase of the dark cycle, “early sessions”) and then again at 10 h into the dark cycle (late phase of the dark cycle, “late sessions”) with an 8-h break between the two sessions in the home cage. Results Adult females consumed significantly more alcohol (g/kg) than males in the 30-min sessions with the FR1 schedule of reinforcement when tested late in the dark cycle. Front-loading of alcohol was the primary factor driving higher consumption in females. Changing the schedule of reinforcement from FR1 to FR3 reduced total consumption. Notably, this manipulation had minimal effect on front-loading behavior in females, whereas front-loading behavior was significantly reduced in males when more effort was required to access alcohol. Compressing drinking access to 15 min to model a happy hour drove up front-loading behavior, generating alcohol drinking patterns in males that were similar to patterns in females (faster drinking and higher intake). Conclusions This strategy could be useful for exploring sex differences in the neural mechanisms underlying alcohol drinking and related health vulnerabilities. Our findings also highlight the importance of the time of testing for detecting sex differences in drinking behavior.

Previous studies demonstrate that Mycobacterium vaccae NCTC 11659 (M. vaccae), a soil-derived bacterium with anti-inflammatory and immunoregulatory properties, is a potentially useful countermeasure against negative outcomes to stressors. Here we used male C57BL/6NCrl mice to determine if repeated immunization with M. vaccae is an effective countermeasure in a “two hit” stress exposure model of chronic disruption of rhythms (CDR) followed by acute social defeat (SD). On day –28, mice received implants of biotelemetric recording devices to monitor 24-h rhythms of locomotor activity. Mice were subsequently treated with a heat-killed preparation of M. vaccae (0.1 mg, administered subcutaneously on days –21, –14, –7, and 27) or borate-buffered saline vehicle. Mice were then exposed to 8 consecutive weeks of either stable normal 12:12 h light:dark (LD) conditions or CDR, consisting of 12-h reversals of the LD cycle every 7 days (days 0–56). Finally, mice were exposed to either a 10-min SD or a home cage control condition on day 54. All mice were exposed to object location memory testing 24 h following SD. The gut microbiome and metabolome were assessed in fecal samples collected on days –1, 48, and 62 using 16S rRNA gene sequence and LC-MS/MS spectral data, respectively; the plasma metabolome was additionally measured on day 64. Among mice exposed to normal LD conditions, immunization with M. vaccae induced a shift toward a more proactive behavioral coping response to SD as measured by increases in scouting and avoiding an approaching male CD-1 aggressor, and decreases in submissive upright defensive postures. In the object location memory test, exposure to SD increased cognitive function in CDR mice previously immunized with M. vaccae. Immunization with M. vaccae stabilized the gut microbiome, attenuating CDR-induced reductions in alpha diversity and decreasing within-group measures of beta diversity. Immunization with M. vaccae also increased the relative abundance of 1-heptadecanoyl-sn-glycero-3-phosphocholine, a lysophospholipid, in plasma. Together, these data support the hypothesis that immunization with M. vaccae stabilizes the gut microbiome, induces a shift toward a more proactive response to stress exposure, and promotes stress resilience.

Abstract Background Patients with autism spectrum disorder (ASD) experience high rates of sleep disruption beginning early in life; however, the developmental consequences of this disruption are not understood. We examined sleep behavior and the consequences of sleep disruption in developing mice bearing C-terminal truncation mutation in the high-confidence ASD risk gene SHANK3 (Shank3ΔC). We hypothesized that sleep disruption may be an early sign of developmental divergence, and that clinically relevant Shank3WT/ΔC mice may be at increased risk of lasting deleterious outcomes following early life sleep disruption. Methods We recorded sleep behavior in developing Shank3ΔC/ΔC, Shank3WT/ΔC, and wild-type siblings of both sexes using a noninvasive home-cage monitoring system. Separately, litters of Shank3WT/ΔC and wild-type littermates were exposed to automated mechanical sleep disruption for 7 days prior to weaning (early life sleep disruption: ELSD) or post-adolescence (PASD) or undisturbed control (CON) conditions. All groups underwent standard behavioral testing as adults. Results Male and female Shank3ΔC/ΔC mice slept significantly less than wild-type and Shank3WT/ΔC siblings shortly after weaning, with increasing sleep fragmentation in adolescence, indicating that sleep disruption has a developmental onset in this ASD model. ELSD treatment interacted with genetic vulnerability in Shank3WT/ΔC mice, resulting in lasting, sex-specific changes in behavior, whereas wild-type siblings were largely resilient to these effects. Male ELSD Shank3WT/ΔC subjects demonstrated significant changes in sociability, sensory processing, and locomotion, while female ELSD Shank3WT/ΔC subjects had a significant reduction in risk aversion. CON Shank3WT/ΔC mice, PASD mice, and all wild-type mice demonstrated typical behavioral responses in most tests. Limitations This study tested the interaction between developmental sleep disruption and genetic vulnerability using a single ASD mouse model: Shank3ΔC (deletion of exon 21). The broader implications of this work should be supported by additional studies using ASD model mice with distinct genetic vulnerabilities. Conclusion Our study shows that sleep disruption during sensitive periods of early life interacts with underlying genetic vulnerability to drive lasting and sex-specific changes in behavior. As individuals progress through maturation, they gain resilience to the lasting effects of sleep disruption. This work highlights developmental sleep disruption as an important vulnerability in ASD susceptibility.

Independent of their beneficial effects on hypertension and cardiovascular related disease, angiotensin receptor type 1 (AT1R) blockers can improve stress-related symptoms. AT1R receptor-mediated actions can be counteracted directly or indirectly by the angiotensin receptor type 2 receptor (AT2R). Our recent studies in a mouse model of PTSD have shown that AT1R blockade increases the extinction (learned inhibition) of a traumatic fear memory and that AT1R mRNA expression is reduced in fear related brain regions of animals treated with the AT1R antagonist losartan. These data imply that downstream AT1 signaling events maybe important in consolidation of fear memory extinction. Therefore we investigated the acute effects of AT2R inhibition and AT2R stimulation on fear memory and baseline anxiety in mice. We performed classical Pavlovian fear conditioning pairing auditory cues with foot shocks and examined fear extinction (% freezing to conditioned stimulus) in the presence of the AT2R antagonist PD 123319 and agonist Compound 21. Twenty-four hours following fear conditioning, PD 123319 (15 mg/kg IP), Compound 21 (10 mg/kg IP) or vehicle was administered prior to fear memory extinction. The PD treated group exhibited significantly less percent freezing (68%; 68 of 100) compared to vehicle control (47%; 47 of 100) during fear expression (F10, 300 = 1.9; p<0.05, n=15) while no effect was observed during extinction retention, an index of long-term fear memory. On the other hand, Compound 21 had no effect on fear expression, extinction or basal levels of anxiety. Moreover, following fear conditioning, qPCR data revealed that mRNA expression of AT2R and angiotensin converting enzyme 2 (ACE2) in the central amygdala were elevated (5 fold and 3 fold respectively, P<0.05, n=6) compared to home cage control, however the AT1R and angiotensin converting enzyme (ACE) gene expression pathways were unaltered. PD 123319 and Compound 21 had no effect on basal levels of anxiety as determined by open field testing. These data indicate that AT1R and AT2R may have divergent effects on short and long-term fear memory formation. Further studies are required to understand the differential regulation of angiotensin receptor signaling in PTSD.

Abstract The onset of appropriate maternal behaviour is essential for the survival of dependent offspring in mammals. Lactogenic hormones, including prolactin and placental lactogen, play an important role in the regulation of this behaviour, mediated through prolactin receptor expression in the medial preoptic area (MPOA) of the hypothalamus1. However, it is unclear how lactogenic action in this region induces the display of maternal behaviour. It has been shown that activation of neuronal projections from the MPOA to the ventral tegmental area (VTA) is necessary for maternal behaviour2, with reward circuitry activated in order to motivate a mother to invest in time- and resource-costly care for her young. We aimed to investigate whether lactogenic hormones play a role in the activation of reward circuitry for maternal behaviour. First, we characterised specific aspects of motivation and reward behaviour in wildtype female C57BL/6 mice of different reproductive states, using three behavioural testing paradigms. We showed that virgin and pregnant mice develop a preference for contexts associated with the presence of foster pups in a conditioned place preference test (p&lt;0.01, n=9 per group), an aspect of reward behaviour that was not observed in lactating females (p=0.14, n=8). However, in a novel T maze and when a climbable barrier was placed in the home cage, virgin and pregnant mice showed low motivation for pup retrieval compared to lactating mice (p&lt;0.05, n≤6 per group). These data demonstrate that reproductive state differentially affects passive reward learning and active motivation for maternal care in wildtype mice. Moreover, we can differentiate between maternal and non-maternal states of pup-related reward behaviour with the use of the T maze and barrier climbing paradigms. Next, we tested mice with a conditional deletion of prolactin receptors in Gamma-Aminobutyric Acid (GABA) neurons (Prlr flox/VGat Cre) in the T maze during lactation. Knockout mice showed incomplete retrieval behaviour, and latencies for pup retrieval were significantly longer than in controls (p&lt;0.01, n≤6 per group). The behavioural impairments observed in Prlr flox/VGat Cre knockout mice in the T maze imply that the action of lactogenic hormones on GABA neurons is required for full maternal motivation. 1.Brown RSE, et al. Prolactin action in the medial preoptic area is necessary for postpartum maternal nursing behavior. Proc Natl Acad Sci U S A114, 10779-10784 (2017). 2.Fang YY, Yamaguchi T, Song SC, Tritsch NX, Lin D. A Hypothalamic Midbrain Pathway Essential for Driving Maternal Behaviors. Neuron98, 192-207 e110 (2018).

As a visual artist, the kitchen has often been the “centre of my home” in a sense that goes beyond the cliché. Over the years I have lived in more than my fair share of tiny houses or apartments where the kitchen has also become my art studio as limited financial resources as well as the needs of my children prevented me from renting a separate studio space. In these kitchen studios, I frequently drew inspiration from things and life around me, sketching aspects of the room and the objects in it. By training a printmaker, these sketches often evolved into prints, usually either as etchings or linocuts. Linocuts particularly lend themselves to working in a limited space as they require little by way of technical equipment. So, after dinner with the dishes cleared away, my linocutting tools and linoblocks would emerge and my kitchen took on a different purpose. Figure 1. Ulrike Sturm. Coffee Pot. 2012. Linocut. 8cm x 11cm. Brian O’Doherty asserts that a different concept of time exists within artist studios where the space is dedicated solely to that purpose. The artist may leave work and tools as they are to come back to later, and in this sense, time is suspended. No such luxury of suspending time is afforded the artist who has to push aside her or his work to prepare the next meal. While many artists work in studios which occupy a domestic space, whether that be the garage, a converted spare bedroom or a basement, there is something about the kitchen that makes its shared function as a food preparation area and a place to make art more contentious, as the tools and equipment of each activity must be completely removed before the alternate function of the space can be implemented. Not least among these considerations are health and safety, which plays a significant role in the need to delineate the functions of the kitchen clearly (Radaydeh and Otoom, Rossol). Brian Winkenweder suggests of the kitchen that “the hermetic reality of this contested space ossifies gender roles and responsibilities in a patriarchal society unlike no other domestic space” (239). Winkenweder’s article focuses on Joseph Beuys and John Cage, and this ossification is apparent in the absence of acknowledgement that this is an issue. However, he suggests that a different perspective is offered by Martha Rosler and Olga Chernysheva, who “confront the kitchen specifically as a women’s space charged with societal tension, dialectically countering romanticized versions of kitchens” (244). In the six minute black and white film of Rosler’s performance piece, Semiotics of the Kitchen, she acts out the politics of the kitchen with deadpan wit. Straight to camera, she performs an alphabet using kitchen implements to demonstrate the letters, often with tense and violent gestures. Rosler’s kitchen can not be viewed as the warm and happy centre of the home: in naming the kitchen objects, the woman is naming her own oppression. Winkenweder examines theoretical implications of the kitchen praxis by both male and female artists and Rosler comments on the politics of the space itself. While acknowledging these dynamics, my interest centres on the shifting dynamic of cooking and making art in the same space. Issues of claiming a space arise and I think of Virginia Woolf’s A Room of One’s Own. I eventually did get a room of my own, so to speak: a dedicated studio space that I love to work in and where the scale of my work is limited by the environment to a much lesser extent. But there are still many evenings when my preferred place to work on my art is at the dining table, with the remnants of dinner around me, and the aromas from the kitchen present. References Bain, Alison L. “Female Artistic Identity in Place: The Studio.” Social & Cultural Geography 5.2 (2004): 171–93. O’Doherty, Brian. Studio and Cube: On The Relationship Between Where Art is Made and Where Art is Displayed. New York: Princeton Architectural P, 2012. Radaydeh, Bassam Naser and Sameer Abd-Alkareem Otoom. “Testing the Awareness of Hazardous Nature of Printmaking Materials among Printmaking Students in Traditional and Non-Toxic Printmaking Programs.” Journal of Health Science 50.6 (2004): 570–5 Rosler, Martha. Semiotics of the Kitchen. Video Data Bank. 1975. 26 May 2013. ‹http://www.vdb.org/titles/semiotics-kitchen›.Rossol, M. The Artist’s Complete Health and Safety Guide. New York: Allworth P, 2001. Winkenweder, Brian. “The Kitchen as Art Studio: Gender, Performance and Domestic Aesthetics.” The Studio Reader: On the Space of Artists. Eds. Mary Jane Jacob and Michelle Grabner. Chicago: U of Chicago P, 2010. 239–50. Woolf, Virginia. A Room of One’s Own. London: L. & V. Woolf , 1929.

Introduction Almost all industries in Australia today have adopted digital media in some way. However, uses in large scale activities such as mining may seem to be different from others. This article looks at mining practices with a media studies approach, and concludes that, just as many other industries, mining and media have converged. Many Australian mine sites are adopting new media for communication and control to manage communication, explore for ore bodies, simulate forces, automate drilling, keep records, and make transport and command robotic. Beyond sharing similar digital devices for communication and computation, new media in mining employ characteristic digital media operations, such as numerical operation, automation and managed variability. This article examines the implications of finding that some of the most material practices have become mediated by new media. Mining has become increasingly mediated through new media technologies similar to GPS, visualisation, game remote operation, similar to those adopted in consumer home and mobile digital media. The growing and diversified adoption of digital media championed by companies like Rio Tinto aims not only ‘improve’ mining, but to change it. Through remediating practices of digital mining, new media have become integral powerful tools in prospective, real time and analytical environments. This paper draws on two well-known case studies of mines in the Pilbara and Western NSW. These have been documented in press releases and media reports as representing changes in media and mining. First, the West Angelas mines in the Pilbara is an open cut iron ore mine introducing automation and remote operation. This mine is located in the remote Pilbara, and is notable for being operated remotely from a control centre 2000km away, near Perth Airport, WA. A growing fleet of Komatsu 930E haul trucks, which can drive autonomously, traverses the site. Fitted with radars, lasers and GPS, these enormous vehicles navigate through the open pit mine with no direct human control. Introducing these innovations to mine sites become more viable after iron ore mining became increasingly profitable in the mid-2000s. A boom in steel building in China drove unprecedented demand. This growing income coincided with a change in public rhetoric from companies like Rio Tinto. They pointed towards substantial investments in research, infrastructure, and accelerated introduction of new media technologies into mining practices. Rio Tinto trademarked the term ‘Mine of the future’ (US Federal News Service 1), and publicised their ambitious project for renewal of mining practice, including digital media. More recently, prices have been more volatile. The second case study site is a copper and gold underground mine at Northparkes in Western NSW. Northparkes uses substantial sensing and control, as well as hybrid autonomous and remote operated vehicles. The use of digital media begins with prospecting, and through to logistics of transportation. Engineers place explosives in optimal positions using computer modelling of the underground rock formations. They make heavy use of software to coordinate layer-by-layer use of explosives in this advanced ‘box cut’ mine. After explosives disrupt the rock layer a kilometre underground, another specialised vehicle collects and carries the ore to the surface. The Sandvik loader-hauler-dumper (LHD) can be driven conventionally by a driver, but it can also travel autonomously in and out of the mine without a direct operator. Once it reaches a collection point, where the broken up ore has accumulated, a user of the surface can change the media mode to telepresence. The human operator then takes control using something like a games controller and multiple screens. The remote operator controls the LHD to fill the scoop with ore. The fully-loaded LHD backs up, and returns autonomously using laser senses to follow a trail to the next drop off point. The LHD has become a powerful mediator, reconfiguring technical, material and social practices throughout the mine. The Meanings of Mining and Media Are Converging Until recently, mining and media typically operated ontologically separately. The media, such as newspapers and television, often tell stories about mining, following regular narrative scripts. There are controversies and conflicts, narratives of ecological crises, and the economics of national benefit. There are heroic and tragic stories such as the Beaconsfield mine collapse (Clark). There are new industry policies (Middelbeek), which are politically fraught because of the lobbying power of miners. Almost completely separately, workers in mines were consumers of media, from news to entertainment. These media practices, while important in their own right, tell nothing of the approaching changes in many other sectors of work and everyday life. It is somewhat unusual for a media studies scholar to study mine sites. Mine sites are most commonly studied by Engineering (Bellamy & Pravica), Business and labour and cultural histories (McDonald, Mayes & Pini). Until recently, media scholarship on mining has related to media institutions, such as newspapers, broadcasters and websites, and their audiences. As digital media have proliferated, the phenomena that can be considered as media phenomena has changed. This article, pointing to the growing roles of media technologies, observes the growing importance that media, in these terms, have in the rapidly changing domain of mining. Another meaning for ‘media’ studies, from cybernetics, is that a medium is any technology that translates perception, makes interpretations, and performs expressions. This meaning is more abstract, operating with a broader definition of media — not only those institutionalised as newspapers or radio stations. It is well known that computer-based media have become ubiquitous in culture. This is true in particular within the mining company’s higher ranks. Rio Tinto’s ambitious 2010 ‘Mine of the Future’ (Fisher & Schnittger, 2) program was premised on an awareness that engineers, middle managers and senior staff were already highly computer literate. It is worth remembering that such competency was relatively uncommon until the late 1980s. The meanings of digital media have been shifting for many years, as computers become experienced more as everyday personal artefacts, and less as remote information systems. Their value has always been held with some ambivalence. Zuboff’s (387-414) picture of loss, intimidation and resistance to new information technologies in the 1980s seems to have dissipated by 2011. More than simply being accepted begrudgingly, the PC platform (and variants) has become a ubiquitous platform, a lingua franca for information workers. It became an intimate companion for many professions, and in many homes. It was an inexpensive, versatile and generalised convergent medium for communication and control. And yet, writers such as Gregg observe, the flexibility of networked digital work imposes upon many workers ‘unlimited work’. The office boundaries of the office wall break down, for better or worse. Emails, utility and other work-related behaviours increasingly encroach onto domestic and public space and time. Its very attractiveness to users has tied them to these artefacts. The trail that leads the media studies discipline down the digital mine shaft has been cleared by recent work in media archaeology (Parikka), platform studies (Middelbeek; Montfort & Bogost; Maher) and new media (Manovich). Each of these redefined Media Studies practices addresses the need to diversify the field’s attention and methods. It must look at more specific, less conventional and more complex media formations. Mobile media and games (both computer-based) have turned out to be quite different from traditional media (Hjorth; Goggin). Kirschenbaum’s literary study of hard drives and digital fiction moves from materiality to aesthetics. In my study of digital mining, I present a reconfigured media studies, after the authors, that reveals heterogeneous media configurations, deserving new attention to materiality. This article also draws from the actor network theory approach and terminology (Latour). The uses of media / control / communications in the mining industry are very complex, and remain under constant development. Media such as robotics, computer modelling, remote operation and so on are bound together into complex practices. Each mine site is different — geologically, politically, and economically. Mines are subject to local and remote disasters. Mine tunnels and global prices can collapse, rendering active sites uneconomical overnight. Many technologies are still under development — including Northparkes and West Angelas. Both these sites are notable for their significant use of autonomous vehicles and remote operated vehicles. There is no doubt that the digital technologies modulate all manner of the mining processes: from rocks and mechanical devices to human actors. Each of these actors present different forms of collusion and opposition. Within a mining operation, the budgets for computerised and even robotic systems are relatively modest for their expected return. Deep in a mine, we can still see media convergence at work. Convergence refers to processes whereby previously diverse practices in media have taken on similar devices and techniques. While high-end PCs in mining, running simulators; control data systems; visualisation; telepresence, and so on may be high performance, ruggedised devices, they still share a common platform to the desktop PC. Conceptual resources developed in Media Ecology, New Media Studies, and the Digital Humanities can now inform readings of mining practices, even if their applications differ dramatically in size, reliability and cost. It is not entirely surprising that some observations by new media theorists about entertainment and media applications can also relate to features of mining technologies. Manovich argues that numerical representation is a distinctive feature of new media. Numbers have always already been key to mining engineering. However, computers visualise numerical fields in simulations that extend out of the minds of the calculators, and into visual and even haptic spaces. Specialists in geology, explosives, mechanical apparatuses, and so on, can use plaftorms that are common to everyday media. As the significance of numbers is extended by computers in the field, more and more diverse sources of data provide apparently consistent and seamless images of multiple fields of knowledge. Another feature that Manovich identifies in new media is the capacity for automation of media operations. Automation of many processes in mechanical domains clearly occurred long before industrial technologies were ported into new media. The difference with new media in mine sites is that robotic systems must vary their performance according to feedback from their extra-system environments. For our purposes, the haul trucks in WA are software-controlled devices that already qualify as robots. They sense, interpret and act in the world based on their surroundings. They evaluate multiple factors, including the sensors, GPS signals, operator instructions and so on. They can repeat the path, by sensing the differences, day after day, even if the weather changes, the track wears away or the instructions from base change. Automation compensates for differences within complex and changing environments. Automation of an open-pit mine haulage system… provides more consistent and efficient operation of mining equipment, it removes workers from potential danger, it reduces fuel consumption significantly reducing greenhouse gas (GHG) emissions, and it can help optimize vehicle repairs and equipment replacement because of more-predictable and better-controlled maintenance. (Parreire and Meech 1-13) Material components in physical mines tend to become modular and variable, as their physical shape lines up with the logic of another of Manovich’s new media themes, variability. Automatic systems also make obsolete human drivers, who previously handled those environmental variations, for better or for worse, through the dangerous, dull and dirty spaces of the mine. Drivers’ capacity to control repeat trips is no longer needed. The Komatsu driverless truck, introduced to the WA iron ore mines from 2008, proved itself to be almost as quick as human drivers at many tasks. But the driverless trucks have deeper advantages: they can run 23 hours each day with no shift breaks; they drive more cautiously and wear the equipment less than human drivers. There is no need to put up workers and their families up in town. The benefit most often mentioned is safety: even the worst accident won’t produce injuries to drivers. The other advantage less mentioned is that autonomous trucks don’t strike. Meanwhile, managers of human labour also need to adopt certain strategies of modulation to support the needs and expectations of their workers. Mobile phones, televisions and radio are popular modes of connecting workers to their loved ones, particularly in the remote and harsh West Angelas site. One solution — regular fly-in-fly out shifts — tends also to be alienating for workers and locals (Cheshire; Storey; Tonts). As with any operations, the cost of maintaining a safe and comfortable environment for workers requires trade-offs. Companies face risks from mobile phones, leaking computer networks, and espionage that expose the site to security risks. Because of such risks, miners tend be subject to disciplinary regimes. It is common to test alcohol and drug levels. There was some resistance from workers, who refused to change to saliva testing from urine testing (Latimer). Contesting these machines places the medium, in a different sense, at the centre of regulation of the workers’ bodies. In Northparkes, the solution of hybrid autonomous and remote operation is also a solution for modulating labour. It is safer and more comfortable, while also being more efficient, as one experienced driver can control three trucks at a time. This more complex mode of mediation is necessary because underground mines are more complex in geology, and working environments to suit full autonomy. These variations provide different relationships between operators and machines. The operator uses a games controller, and watches four video views from the cabin to make the vehicle fill the bucket with ore (Northparkes Mines, 9). Again, media have become a pivotal element in the mining assemblage. This combines the safety and comfort of autonomous operation (helping to retain staff) with the required use of human sensorimotor dexterity. Mine systems deserve attention from media studies because sites are combining large scale physical complexity with increasingly sophisticated computing. The conventional pictures of mining and media rarely address the specificity of subjective and artefactual encounters in and around mine sites. Any research on mining communication is typically within the instrumental frames of engineering (Duff et al.). Some of the developments in mechanical systems have contributed to efficiency and safety of many mines: larger trucks, more rock crushers, and so on. However, the single most powerful influence on mining has been adopting digital media to control, integrate and mining systems. Rio Tinto’s transformative agenda document is outlined in its high profile ‘Mine of the Future’ agenda (US Federal News Service). The media to which I refer are not only those in popular culture, but also those with digital control and communications systems used internally within mines and supply chains. The global mining industry began adopting digital communication automation (somewhat) systematically only in the 1980s. Mining companies hesitated to adopt digital media because the fundamentals of mining are so risky and bound to standard procedures. Large scale material operations, extracting and processing minerals from under the ground: hardly to be an appropriate space for delicate digital electronics. Mining is also exposed to volatile economic conditions, so investing in anything major can be unattractive. High technology perhaps contradicts an industry ethos of risk-taking and masculinity. Digital media became domesticated, and familiar to a new generation of formally educated engineers for whom databases and algorithms (Manovich) were second nature. Digital systems become simultaneously controllers of objects, and mediators of meanings and relationships. They control movements, and express communications. Computers slide from using meanings to invoking direct actions over objects in the world. Even on an everyday scale, computer operations often control physical processes. Anti-lock Braking Systems regulate a vehicle’s braking pressure to avoid the danger when wheels lock-up. Or another example, is the ATM, which involves both symbolic interactions, and also exchange of physical objects. These operations are examples of the ‘asignifying semiotic’ (Guattari), in which meanings and non-meanings interact. There is no operation essential distinction between media- and non-media digital operations. Which are symbolic, attached or non-consequential is not clear. This trend towards using computation for both meanings and actions has accelerated since 2000. Mines of the Future Beyond a relatively standard set of office and communications software, many fields, including mining, have adopted specialised packages for their domains. In 3D design, it is AutoCAD. In hard sciences, it is custom modelling. In audiovisual production, it may be Apple and Adobe products. Some platforms define their subjectivity, professional identity and practices around these platforms. This platform orientation is apparent in areas of mining, so that applications such as the Gemcom, Rockware, Geological Database and Resource Estimation Modelling from Micromine; geology/mine design software from Runge, Minemap; and mine production data management software from Corvus. However, software is only a small proportion of overall costs in the industry. Agents in mining demand solutions to peculiar problems and requirements. They are bound by their enormous scale; physical risks of environments, explosive and moving elements; need to negotiate constant change, as mining literally takes the ground from under itself; the need to incorporate geological patterns; and the importance of logistics. When digital media are the solution, there can be what is perceived as rapid gains, including greater capacities for surveillance and control. Digital media do not provide more force. Instead, they modulate the direction, speed and timing of activities. It is not a complete solution, because too many uncontrolled elements are at play. Instead, there are moment and situations when the degree of control refigures the work that can be done. Conclusions In this article I have proposed a new conception of media change, by reading digital innovations in mining practices themselves as media changes. This involved developing an initial reading of the operations of mining as digital media. With this approach, the array of media components extends far beyond the conventional ‘mass media’ of newspapers and television. It offers a more molecular media environment which is increasingly heterogeneous. It sometimes involves materiality on a huge scale, and is sometimes apparently virtual. The mining media event can be a semiotic, a signal, a material entity and so on. It can be a command to a human. It can be a measurement of location, a rock formation, a pressure or an explosion. The mining media event, as discussed above, is subject to Manovich’s principles of media, being numerical, variable and automated. In the mining media event, these principles move from the aesthetic to the instrumental and physical domains of the mine site. The role of new media operates at many levels — from the bottom of the mine site to the cruising altitude of the fly-in-fly out aeroplanes — has motivated significant changes in the Australian industry. When digital media and robotics come into play, they do not so much introduce change, but reintroduce similarity. This inversion of media is less about meaning, and more about local mastery. Media modulation extends the kinds of influence that can be exerted by the actors in control. In these situations, the degrees of control, and of resistance, are yet to be seen. Acknowledgments Thanks to Mining IQ for a researcher's pass at Mining Automation and Communication Conference, Perth in August 2012. References Bellamy, D., and L. 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