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The use of head fixation in mice is increasingly common in research, its use having initially been restricted to the field of sensory neuroscience. Head restraint has often been combined with fluid control, rather than food restriction, to motivate behaviour, but this too is now in use for both restrained and non-restrained animals. Despite this, there is little guidance on how best to employ these techniques to optimise both scientific outcomes and animal welfare. This article summarises current practices and provides recommendations to improve animal wellbeing and data quality, based on a survey of the community, literature reviews, and the expert opinion and practical experience of an international working group convened by the UK's National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs). Topics covered include head fixation surgery and post-operative care, habituation to restraint, and the use of fluid/food control to motivate performance. We also discuss some recent developments that may offer alternative ways to collect data from large numbers of behavioural trials without the need for restraint. The aim is to provide support for researchers at all levels, animal care staff, and ethics committees to refine procedures and practices in line with the refinement principle of the 3Rs.

Trunk shakers are widely used for olive harvesting, being the main detachment system for fruit harvesting. In recent decades, the components of trunk shakers have evolved at mechanical, hydraulic and control levels. However, machine accuracy depends on the operator, whose expertise is a key factor for issues such as trunk debarking caused by grabbing systems, shaking parameters, or on-foot operator safety. The objective of this work was to develop an automatic trunk-detection system to reduce operator influence on the process. Thus, the automatic system via infrared sensor was implemented on a trunk shaker head hitched to a tractor. A control algorithm, control logic and display for trunk grabbing automation were developed. The automatic system was tested under laboratory and field conditions to assess the influence of some variables on trunk detection. The evaluated variables were colour, material, diameter, and target location within the sensor field of vision. The success rate of the automatic system was 91% for trunk grabbing. In the field phase, the efficacy of the automatic system was compared to an operator performing the tasks manually, obtaining times of 16.05 ± 2.8 s tree−1 and 21.54 ± 5.29 s tree−1 respectively, and a percentage of success in trunk grabbing of 92.9%. Automatic mode improved manual mode by saving 27.3% of time, improving effective field capacity. The automatic mode developed here provided a high ratio of success and it showed highly reliable and efficient performance compared with manual mode.

Summary Optogenetic tools and imaging methods for recording and manipulating brain activity have boosted the field of neuroscience in unprecedented ways. However, behavioral paradigms for mice lag behind those of primates, limiting the full potential of such tools. Here, we present an innovative behavioral framework in which head-fixed mice directionally reach for water droplets, similar to the primate “center-out” reaching task. Mice rapidly engaged in the task, performed hundreds of trials, and reached in multiple directions when droplets were presented at different locations. Surprisingly, mice used chemosensation to determine the presence of water droplets. Optogenetic inactivation of the motor cortex halted the initiation and rapidly diverted the trajectory of ongoing movements. Layer 2/3 two-photon imaging revealed robust direction selectivity in most reach-related neurons. Finally, mice performed directional reaching instructed by vibratotactile stimuli, demonstrating the potential of this framework for studying, in addition to motor control, sensory processing, and decision making., Graphical Abstract, Highlights • Mice rapidly learn to reach for water droplets in multiple directions using chemosensation • Directional reaching can be instructed by arbitrary sensorimotor associations • Optogenetic motor cortex silencing halts movement initiation and ongoing reaching • Motor cortex layer 2/3 neurons display a high degree of directional selectivity, Galiñanes et al. present a behavioral framework for systems neuroscience in which mice learn to reach for water droplets presented at multiple locations, similar to the primate center-out task. Reaching is guided by chemosensory cues and depends on motor cortex. Layer 2/3 neurons display a high degree of directional selectivity.

Olive harvesting and pruning are two operations that highly influenced olive growing costs and competitiveness. Both operations should be related to reach an efficient orchard management. Tree structure should be adapted to selected harvesting method, but it also has to be in accordance with tree and orchard features. This research attempts to assess how traditional olives could be adapted to harvesting with canopy shakers, measuring different parameters for pruning and harvester performance. For this purpose, two pruning tests were conducted in 'Picual' and 'Hojiblanca' traditional olive orchards during four years. Three pruning treatments were pruned to adapt tree canopy to different harvesting systems. Harvesting was performed using two different canopy shakers. Harvesting efficiency and debris production were measured as descriptive variables to analyze harvester performance. On the one hand, pruning fresh weight was highly variable between years, however trunk shaker pruning provided higher amount of yearly pruning dry weight while canopy shaker pruning generated higher accumulated pruning dry weight. On the other hand, canopy shaker pruning provided higher harvesting efficiency than other pruning systems for pre-commercial harvester. With regard to debris production, it was highly variable, although it provided significant differences between both cultivars. Finally, pruning and harvesting operations should be related to choose an efficient harvesting system and to adapt the trees to the harvester and vice versa. This rule should be taken into account to carry out an adequate mechanization of an olive orchard.

Because of the pollution caused by inappropriate pesticide applications to olive canopies, the Mecaolivar pre-commercial procurement project was undertaken to develop new airblast sprayers to optimise application efficiency and overcome the limitations of conventional sprayers used in traditional and intensive orchards. Three prototype sprayers were developed, evaluated, and calibrated under laboratory conditions and were tested in the field by spraying trees in traditional and intensive cultivation systems. Water-sensitive paper was used to assess the spray coverage achieved. The prototype sprayers were designed to adapt the deposition nozzle positions to the canopy shape to reduce spray drift and off-target application. The first prototype (P1) consisted of a sprayer with a centrifugal fan and adaptable individual spouts, the second (P2) consisted of a sprayer with six small hydraulically-driven axial fans mounted on two mobile structures, and the third (P3) consisted of two axial fans mounted on a tower-like structure with mobile air outlets. The results of the field test showed that the prototypes could be more efficient than conventional equipment. In applying the same liquid volume, the P2 and P3 prototypes increased the coverage by 61% and 46% on average in intensive and traditional systems, respectively, compared to a commercial airblast sprayer, without a significant decrease in the deposit homogeneity throughout the crown.

Citrus mechanical harvesting has been investigated since the 1960's. Even though mechanical harvesting could significantly lower production costs, the implementation by the private sector has been slow. The current harvesting technologies detach the fruits with trunk, canopy or branch vibration. For late-season sweet orange varieties which simultaneously bear mature fruit, immature fruitlets and flowers, shaker harvesting decreases the subsequent year's yield. This study, investigated the frequency response of mature fruits and immature fruitlets to determine the optimum frequency range for an efficient and selective harvest. Laboratory vibration transmission tests were conducted with 14 branches bearing 76 mature fruits and 151 immature ‘Valencia’ fruitlets. The fruit and branch response to the forced vibration was measured by several sets of five triaxial accelerometers with a dynamic signal analyser. Three frequency ranges with the highest vibration transmission values were identified for mechanical harvesting lower than 10 Hz. The first frequency range (1.5–2.5 Hz) corresponded best with the most efficient vibration transmission, involving more than 90% of fruit. The second frequency range (4.5–5 Hz) successfully discriminated between mature fruit and immature fruitlets. In this frequency range, 53.4% of mature fruit amplified the acceleration a mean value of 2.2 times, while only 7.3% of immature fruitlets amplified the acceleration with a mean value of 4.4 times. The third frequency range (7–8 Hz) had the lowest vibration transmission value. The frequency response of mature citrus fruits, and their markedly higher fruit mass, were significant factors in efficient selective mechanical harvesting.

The interest in conservation agriculture in Spain is evidenced by practical and institutional aspects. The practical aspect is reflected by the area cultivated under this farming system, 1.28 Mha in perennial crops and 0.57 Mha in arable crops, both for 2013. The period under review was 2009–2013 for arable crops and 2006–2013 for perennial crops. In that period, figures increased 208% for no tillage in arable crops, and 54% for groundcovers in perennial crops. The institutional support is reflected by the financial funding given to conservation agriculture farming practices by some Spanish Regional Governments, primarily through Rural Development Programs, that reached over € 200 million in the 2000–2006 period. The origins of soil conservation practices date back to the 1930s and have evolved in parallel in America and Europe. This parallelism has led to the use of different terminology for similar practices that do not always fall within the scope of conservationist practices. Consistent with the literature, and based on the results of 6 meetings with 144 Spanish experts, this paper aimed at clarifying terms and practices applied under the conditions of Spain, but could be useful for other geographies. This article also proposes definitions to clearly describe the different concepts for experts, advisers, and also for policy makers to accurately allocate funds in the European financial framework 2014–2020.

Low harvesting efficiency and fruit damage are the main limitations for trunk shakers in the harvesting of table olives. Different cultivations of olive orchards, variations in the fruit properties over the harvesting season, and a wide range of machines both characterise and hamper the harvesting process in several traditional olive producing countries, such as Spain. The aim of the work reported was to develop the mechanical harvesting of existing orchards using commercial trunk shakers maximising the quantity of harvested fruit while maintaining its quality. Results showed that the mean value of harvesting efficiency with trunk shakers was 74%, when additional rod beating or abscission agents were not applied. In orchards where the trees were trained into shapes that facilitated the transmission of vibration, fruit properties had a limited effect on harvesting efficiency. In order to achieve harvesting efficiency >85%, tree trunk vibration parameters were set above an acceleration value of 183.4 m s−2, and at a frequency of 28.1 Hz. Although increasing the trunk acceleration improved harvesting efficiency, it led to an increase in damage to the harvested fruit. Damage caused to the fruit through mechanical harvesting was 3.5 times greater than the damage caused by manual harvesting. Adjusting the vibration parameters, limiting the power of the machine, training the canopy to reduce volume and facilitating the vibration transmission to the fruit, together with effective post-harvest field treatments that maintain the fruit quality, are all required to ensure the success of vibration harvesting systems for green olives.

Most of the advances made in olive traceability are focusing on the arrival of the fruit to industry but it is necessary to manage all the operations involved in the production chain. This work introduces a new machine-to-machine system that allows integrating all the information generated from the field to the market through a methodology for the real-time management of all operations carried out. The consumer can check the product history, farmers can consult and optimise their resources, and industry can control the operations performed. The system developed is composed of an electronic device, iOlivetrack-D, mounted on agricultural machinery which identify plots and sends the information generated to a web application, iOlivetrack-W. The system was tested in real working conditions obtaining good results in the identification of plots using RFID and GPS although the limitation about this technology must be considered for a commercial purpose. The registration of information associated in the web application has been carried successfully by an adaptable system that allow the access to the data in configurable ways. To complete the product chain information, industrial processing operations were simulated by entering the data manually through the application. Finally, a QR code was generated to provide consumer access to product traceability information that may be consulted in this paper. The work shows the pros and cons of this system which allows assured traceability of the entire production chain and the proper manage of the production.

Developing mechanical harvesting for table olives will require decreasing fruit damage during harvest and postharvest handling, transport and storage. The susceptibility to bruising and its development over time were studied in three table olive varieties, cv. ‘Manzanilla’, ‘Gordal Sevillana’ and ‘Hojiblanca’. Bruising was produced with controlled energy impacts of 56, 26, 13 mJ. A strong correlation ( r 2 = 0.77–0.90) between bruise volume and impact energy was demonstrated. Bruise susceptibility was higher in the Manzanilla variety, followed by Hojiblanca and Gordal Sevillana cultivars. Bruise time evolution was evaluated using a spectrophotometer for visible and near infrared regions. A bruise index was developed using different wavelengths, 545, 670 and 800 nm. Most darkening due to the browning process happened within 1 h, was exponential and dependent on impact energy level. The discoloration was greatest in the Manzanilla, followed by Hojiblanca and Gordal Sevillana olives.

Bruise damage on table olives is the main factor that reduces fruit quality and leads to a severe loss of product during harvesting and postharvest handling operations. The Manzanilla cultivar is the most important table olive variety and it is also very susceptible to the formation of bruises. In this study, visible and near-infrared spectroscopy techniques were used to detect bruise damage on fruit with different absorbed impact energy levels (low = 13 ± 2 mJ, medium = 27 ± 2 mJ and high = 58 ± 7 mJ). The visible spectral region of 535–680 nm was used to distinguish between undamaged and damaged fruit for qualitative analysis. The greater differences in reflectance in the near-infrared region of 700–950 nm enabled a good quantitative analysis by distinguishing between the different impact energy levels. Modified partial least square models were developed to determine the bruise volume in damaged fruits and the absorbed energy during impact. Good fits (r2 = 0.87–0.90) were obtained between the values predicted by visible and near infrared spectroscopy and the values measured in laboratory by the reference methods for bruise volume and absorbed impact energy.

Mechanical harvesting of the stone pine ( Pinus pinea L.) with trunk shakers is replacing manual harvesting. The machines used are generally involved in the mechanical harvesting of agricultural species; their adjustments and use are not adapted to the needs of forest species. The objective of this study was to analyse the vibration process and maximise the number of mature cones harvested whilst limiting any potential damage caused to the tree. Tests were performed in two stone pine plantations located in southern Spain on two harvest dates. The vibration parameters, the harvest efficiency of mature cones and the damage caused to the tree by the loss of shoots, immature cones and damage to the bark were analysed. Adjusting the trunk shaker in a frequency range of 16.1–18.9 Hz and the effective acceleration values at the trunk in the range 51.2–78.4 m s −2 removed an average value of 85.7% of the mature cones from the tree. With these parameters, the average damage caused to the tree was less than two non-harvestable organs per tree. The harvest date was a fundamental parameter in limiting the damage caused to the tree and bark, avoiding vibration after the start of development of new shoots. Vibration periods of more than 4 s were not effective in increasing the number of mature cones removed. Tree vibrations with two short-duration vibrations were effective in increasing the amount of mature cones removed without increasing the damage caused to the tree.

Up states are a hallmark of striatal physiology. Spontaneous activity in the thalamo-cortical network drives robust plateau depolarizations in the medium spiny projection neurons of the striatum. Medium spiny neuron firing is only possible during up states and is very tightly regulated by dopamine and NMDA receptors. In a rat model of Parkinson's disease the medium spiny neurons projecting to the globus pallidus (indirect pathway) show more depolarized up states and increased firing. This is translated into abnormal patterns of synchronization between the globus pallidus and frontal cortex, which are believed to underlie the symptoms of Parkinson's disease. Here we review our work in the field and propose a mechanism through which the lack of D2 receptor stimulation in the striatum allows the establishment of fixed routes of information flow in the cortico-striato-pallidal network.

The objectives of this study were: (1) to investigate, in a clinical practice setting, the effectiveness of olanzapine in the treatment of schizophrenia among partially-responding, symptomatic Asian patients who switch from conventional antipsychotic treatment, (2) to assess the safety of olanzapine and (3) to assess the change in quality of life in Asian patients with schizophrenia who switch to olanzapine.Effectiveness, safety and quality of life were assessed in outpatients with schizophrenia (n=1267) who lacked symptomatic control with conventional antipsychotics and were switched to olanzapine therapy. Data for this prospective, observational study were collected for 12 months from Asian patients in China, Hong Kong, the Philippines, South Korea and Taiwan.Significant clinical improvements (P0.05) were observed following 12 months of olanzapine treatment and 87.3% of the subjects responded to treatment at endpoint (i.e. Brief Psychiatric Rating Scale Total score reduced byor =30% relative to baseline; last observation carried forward). Abnormal involuntary movements (mean change in Abnormal Involuntary Movement Scale: -3.20, P0.001) and quality of life were significantly improved in patients treated with olanzapine. However, some patients experienced significant weight gain (3.60+/-4.50 kg, P0.001) with olanzapine treatment, relative to baseline.This study shows that switching to olanzapine may be effective in improving symptoms, may be well-tolerated and may improve the quality of life in Asian patients who are only partially responsive to treatment with conventional antipsychotics. The pragmatic design and naturalistic setting of this large study make the findings relevant for treating patients from some Asian countries in routine clinical practice.

Summary Neuronal motor commands, whether generating real or neuroprosthetic movements, are shaped by ongoing sensory feedback from the displacement being produced. Here we asked if cortical stimulation could provide artificial feedback during operant conditioning of cortical neurons. Simultaneous two-photon imaging and real-time optogenetic stimulation were used to train mice to activate a single neuron in motor cortex (M1), while continuous feedback of its activity level was provided by proportionally stimulating somatosensory cortex. This artificial signal was necessary to rapidly learn to increase the conditioned activity, detect correct performance, and maintain the learned behavior. Population imaging in M1 revealed that learning-related activity changes are observed in the conditioned cell only, which highlights the functional potential of individual neurons in the neocortex. Our findings demonstrate the capacity of animals to use an artificially induced cortical channel in a behaviorally relevant way and reveal the remarkable speed and specificity at which this can occur., Highlights • All-optical brain-machine-brain interface for neuroprosthetic control • Mice rapidly learn to activate single neurons under optogenetically evoked feedback • Population imaging reveals that learning is restricted to the conditioned neuron • Novel “in cerebro” learning paradigm for neural circuit dissection, Prsa et al. demonstrate that cortical stimulation can provide informative feedback about the current state of a voluntarily modulated neuron. Rapid learning of this artificial paradigm holds promise for its use as a research tool and in neuroprosthetic applications.

PRA1 (prenylated Rab acceptor) is a general regulator of Rab proteins, while RILP (Rab interacting lysosomal protein) is a specific effector for Rab7. It has been shown that PRA1 interacts with Rab proteins and with VAMP2. Therefore PRA1 is probably an important factor for membrane traffic, linking together the function of Rab proteins and SNAREs. RILP has a key role in the control of transport to degradative compartments together with Rab7 and probably links Rab7 function to the cytoskeleton. Here we have studied by Northern blot the expression of the two genes in several different human tissues. The 0.8-kb mRNA for human PRA1 is ubiquitously expressed, while the two mRNAs for RILP are differentially expressed. In addition, we have assigned the human PRA1 gene to chromosome 19q13.13-q13.2 and the human RILP gene to chromosome 17p13.3.