Analyze Diet

Scientific reports.

Periodical
Science
Natural Science Disciplines
Publisher:
Nature Publishing Group,
Frequency: Continuously updated
Country: England
Language: English
Start Year:2011 -
ISSN:
2045-2322 (Electronic)
2045-2322 (Linking)
Impact Factor
4.6
2022
NLM ID:101563288
(OCoLC):732869387
LCCN:2011250880
Identification of the novel polymorphisms and potential genetic features of the prion protein gene (PRNP) in horses, a prion disease-resistant animal.
Scientific reports    June 2, 2020   Volume 10, Issue 1 8926 doi: 10.1038/s41598-020-65731-5
Kim YC, Won SY, Do K, Jeong BH.Prion diseases, a protein misfolded disorder (PMD) caused by misfolded prion protein (PrP), present in a wide variety of hosts, ranging from ungulates to humans. To date, prion infections have not been reported in horses, which are well-known as prion disease-resistant animals. Several studies have attempted to identify distinctive features in the prion protein of horses compared to prion disease-susceptible animals, without the study on polymorphisms of the horse prion protein gene (PRNP). Since single nucleotide polymorphisms (SNPs) of PRNP in prion disease-susceptible animals are major susc...
Priming for welfare: gut microbiota is associated with equitation conditions and behavior in horse athletes.
Scientific reports    May 20, 2020   Volume 10, Issue 1 8311 doi: 10.1038/s41598-020-65444-9
Mach N, Ruet A, Clark A, Bars-Cortina D, Ramayo-Caldas Y, Crisci E, Pennarun S, Dhorne-Pollet S, Foury A, Moisan MP, Lansade L.We simultaneously measured the fecal microbiota and multiple environmental and host-related variables in a cohort of 185 healthy horses reared in similar conditions during a period of eight months. The pattern of rare bacteria varied from host to host and was largely different between two time points. Among a suite of variables examined, equitation factors were highly associated with the gut microbiota variability, evoking a relationship between gut microbiota and high levels of physical and mental stressors. Behavioral indicators that pointed toward a compromised welfare state (e.g. stereotyp...
Equine penile squamous cell carcinoma: expression of biomarker proteins and EcPV2.
Scientific reports    May 12, 2020   Volume 10, Issue 1 7863 doi: 10.1038/s41598-020-64014-3
Arthurs C, Suarez-Bonnet A, Willis C, Xie B, Machulla N, Mair TS, Cao K, Millar M, Thrasivoulou C, Priestnall SL, Ahmed A.Equine penile squamous cell carcinoma (EpSCC) is a relatively common cutaneous neoplasm with a poor prognosis. In this study, we aimed to determine the protein expression and colocalisation of FRA1, c-Myc, Cyclin D1, and MMP7 in normal (NT), tumour (T), hyperplastic epidermis and/or squamous papilloma (Hyp/Pap), poorly-differentiated (PDSCC), or well-differentiated (WDSCC) EpSCC using a tissue array approach. Further objectives were to correlate protein expression to (i) levels of inflammation, using a convolutional neural network (ii) equine papillomavirus 2 (EcPV2) infection, detected using ...
Efficient correction of a deleterious point mutation in primary horse fibroblasts with CRISPR-Cas9.
Scientific reports    May 4, 2020   Volume 10, Issue 1 7411 doi: 10.1038/s41598-020-62723-3
Pinzon-Arteaga C, Snyder MD, Lazzarotto CR, Moreno NF, Juras R, Raudsepp T, Golding MC, Varner DD, Long CR.Phenotypic selection during animal domestication has resulted in unwanted incorporation of deleterious mutations. In horses, the autosomal recessive condition known as Glycogen Branching Enzyme Deficiency (GBED) is the result of one of these deleterious mutations (102C > A), in the first exon of the GBE1 gene (GBE1). With recent advances in genome editing, this type of genetic mutation can be precisely repaired. In this study, we used the RNA-guided nuclease CRISPR-Cas9 (clustered regularly-interspaced short palindromic repeats/CRISPR-associated protein 9) to correct the GBE1 mutation i...
Detection of Neorickettsia risticii, the agent of Potomac horse fever, in horses from Rio de Janeiro, Brazil.
Scientific reports    April 29, 2020   Volume 10, Issue 1 7208 doi: 10.1038/s41598-020-64328-2
Paulino PG, Almosny N, Oliveira R, Viscardi V, Müller A, Guimarães A, Baldani C, da Silva C, Peckle M, Massard C, Santos H.This study aims to report the presence of Neorickettsia risticii DNA in blood samples from naturally infected horses in Rio de Janeiro, provide clinicopathological findings related to the infection, and report the phylogenetic diversity of the 16S rDNA of N. risticii in order to evaluate its heterogeneity. Real-time quantitative polymerase chain reaction (qPCR) was performed to investigate the presence of N. risticii in samples collected from horses (n = 187). Five positive samples were found in the molecular screening. Hypoalbuminemia and high levels of creatine kinase and lactate dehydro...
The complexity of clinically-normal sinus-rhythm ECGs is decreased in equine athletes with a diagnosis of paroxysmal atrial fibrillation.
Scientific reports    April 22, 2020   Volume 10, Issue 1 6822 doi: 10.1038/s41598-020-63343-7
Alexeenko V, Fraser JA, Bowen M, Huang CL, Marr CM, Jeevaratnam K.Equine athletes have a pattern of exercise which is analogous to human athletes and the cardiovascular risks in both species are similar. Both species have a propensity for atrial fibrillation (AF), which is challenging to detect by ECG analysis when in paroxysmal form. We hypothesised that the proarrhythmic background present between fibrillation episodes in paroxysmal AF (PAF) might be detectable by complexity analysis of apparently normal sinus-rhythm ECGs. In this retrospective study ECG recordings were obtained during routine clinical work from 82 healthy horses and from 10 horses with a ...
Modulation of stress and immune response by Amblyomin-X results in tumor cell death in a horse melanoma model.
Scientific reports    April 14, 2020   Volume 10, Issue 1 6388 doi: 10.1038/s41598-020-63275-2
Lichtenstein F, Iqbal A, de Lima Will SEA, Bosch RV, DeOcesano-Pereira C, Goldfeder MB, Chammas R, Trufen CEM, Morais KLP, de Souza JG, Natalino RJM....We have investigated Amblyomin-X-treated horse melanomas to better understand its mode of action through transcriptome analysis and the in vivo model. Amblyomin-X is a Kunitz-type homologous protein that selectively leads to the death of tumor cells via ER stress and apoptosis, currently under investigation as a new drug candidate for cancer treatment. Melanomas are immunogenic tumors, and a better understanding of the immune responses is warranted. Equine melanomas are spontaneous and not so aggressive as human melanomas are, as this study shows that the in vivo treatment of encapsulated hors...
Female horses spontaneously identify a photograph of their keeper, last seen six months previously.
Scientific reports    April 14, 2020   Volume 10, Issue 1 6302 doi: 10.1038/s41598-020-62940-w
Lansade L, Colson V, Parias C, Trösch M, Reigner F, Calandreau L.Horses are capable of identifying individual conspecifics based on olfactory, auditory or visual cues. However, this raises the questions of their ability to recognize human beings and on the basis of what cues. This study investigated whether horses could differentiate between a familiar and unfamiliar human from photographs of faces. Eleven horses were trained on a discrimination task using a computer-controlled screen, on which two photographs were presented simultaneously (32 trials/session): touching one was rewarded (S+) and the other not (S-). In the training phase, the S+ faces were of...
Author Correction: Spotted phenotypes in horses lost attractiveness in the Middle Ages.
Scientific reports    April 10, 2020   Volume 10, Issue 1 6469 doi: 10.1038/s41598-020-62905-z
Wutke S, Benecke N, Sandoval-Castellanos E, Döhle HJ, Friederich S, Gonzalez J, Hallsson JH, Hofreiter M, Lõugas L, Magnell O, Morales-Muniz A....An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Rapid isothermal duplex real-time recombinase polymerase amplification (RPA) assay for the diagnosis of equine piroplasmosis.
Scientific reports    March 5, 2020   Volume 10, Issue 1 4096 doi: 10.1038/s41598-020-60997-1
Lei R, Wang X, Zhang D, Liu Y, Chen Q, Jiang N.Equine piroplasmosis (EP) is a severe disease of horses caused by the tick-borne protozoa Theileria equi (T. equi) and Babesia caballi (B. caballi). Infectious carriers are not always symptomatic, meaning there is a risk to non-enzootic areas. Regulatory tests for EP include sero-epidemiological methods for equine babesiosis, but these lack specificity due to cross-reactivity with other Babesia species. In this study, we present a real-time quantitative recombinase polymerase amplification (qRPA) method for fast simultaneous detection of both T. equi and B. caballi. In this method, primers and...
Molecular sexing and preliminary assessment of population sex ratio of the endangered Malayan tapir (Tapirus indicus) in Peninsular Malaysia.
Scientific reports    March 4, 2020   Volume 10, Issue 1 3973 doi: 10.1038/s41598-020-60552-y
Lim QL, Tan YL, Ng WL, Yong CSY, Ismail A, Rovie-Ryan JJ, Rosli N, Annavi G.A molecular sexing method by polymerase chain reaction (PCR) amplification of a portion of the sex-determining region Y (SRY) and the zinc finger (ZF) gene, as well as six equine Y-chromosome-specific microsatellite markers, were tested in the Malayan tapir (Tapirus indicus). While the microsatellite markers did not yield any male-specific amplicons for sex-typing, the SRY/ZF marker system produced reliable molecular sexing results by accurately sex-typing 31 reference Malayan tapirs, using whole blood, dried blood spot (DBS), or tissue samples as materials for DNA extraction. The marker syste...
Energy metabolism of the equine cumulus oocyte complex during in vitro maturation.
Scientific reports    February 26, 2020   Volume 10, Issue 1 3493 doi: 10.1038/s41598-020-60624-z
Lewis N, Hinrichs K, Leese HJ, McG Argo C, Brison DR, Sturmey R.Horses are one of the few species, beside humans, in which assisted reproductive technology has important clinical applications. Furthermore, the horse can serve as a valuable model for the study of comparative reproductive biology. Here we present the first comprehensive characterisation of energy metabolism and mitochondrial efficiency in equine cumulus-oocyte complexes (COCs) during in vitro maturation (IVM), as determined using a combination of non-invasive consumption and release assays and mitochondrial function analysis. These data reveal notable species-specific differences in the rate...
Spread of equine arteritis virus among Hucul horses with different EqCXCL16 genotypes and analysis of viral quasispecies from semen of selected stallions.
Scientific reports    February 19, 2020   Volume 10, Issue 1 2909 doi: 10.1038/s41598-020-59870-y
Socha W, Sztromwasser P, Dunowska M, Jaklinska B, Rola J.Equine arteritis virus (EAV) is maintained in the horse populations through persistently infected stallions. The aims of the study were to monitor the spread of EAV among Polish Hucul horses, to analyse the variability of circulating EAVs both between- and within-horses, and to identify allelic variants of the serving stallions EqCXCL16 gene that had been previously shown to strongly correlate with long-term EAV persistence in stallions. Serum samples (n = 221) from 62 horses including 46 mares and 16 stallions were collected on routine basis between December 2010 and May 2013 and tested f...
Bit type exerts an influence on self-controlled rein tension in unridden horses.
Scientific reports    February 12, 2020   Volume 10, Issue 1 2420 doi: 10.1038/s41598-020-59400-w
Kau S, Potz IK, Pospisil K, Sellke L, Schramel JP, Peham C.Bit configuration and acting rein forces play a crucial role in oral health and comfort of ridden horses. Although it is a big animal welfare issue, dynamic response of horses to different bits has yet not been thoroughly investigated. This convenience sample experimental study describes a model to overcome the almost uncontrollable influence of riders on rein tension and evaluates self-controlled maximum side rein tension of ten sound horses randomly bitted with a double-jointed (DJS) and a version of a Mullen mouth snaffle-bit under unridden conditions. Horses were exercised at walk and trot...
The oxytocin-prostaglandins pathways in the horse (Equus caballus) placenta during pregnancy, physiological parturition, and parturition with fetal membrane retention.
Scientific reports    February 7, 2020   Volume 10, Issue 1 2089 doi: 10.1038/s41598-020-59085-1
Rapacz-Leonard A, Leonard M, Chmielewska-Krzesińska M, Siemieniuch M, Janowski TE.Despite their importance in mammalian reproduction, substances in the oxytocin-prostaglandins pathways have not been investigated in the horse placenta during most of pregnancy and parturition. Therefore, we quantified placental content of oxytocin (OXT), oxytocin receptor (OXTR), and prostaglandin E2 and F2 alpha during days 90-240 of pregnancy (PREG), physiological parturition (PHYS), and parturition with fetal membrane retention (FMR) in heavy draft horses (PREG = 13, PHYS = 11, FMR = 10). We also quantified OXTR and prostaglandin endoperoxide synthase-2 (PTGS2) mRNA expression ...
Matrix metallopeptidase expression and modulation by transforming growth factor-β1 in equine endometrosis.
Scientific reports    January 24, 2020   Volume 10, Issue 1 1119 doi: 10.1038/s41598-020-58109-0
Szóstek-Mioduchowska A, Słowińska M, Pacewicz J, Skarzynski DJ, Okuda K.Equine endometrial fibrosis (endometrosis) is described as a degenerative chronic condition in the uterus. Its characteristic feature is excessive deposition of extracellular matrix (ECM) components around the endometrial glands and stroma. Although matrix metallopeptidases (MMPs) that mediate ECM turnover are important factors in the process of fibrosis, knowledge of their expression and regulation in endometrosis is limited. In other species, one of the important regulators of MMPs and tissue inhibitors of MMPs (TIMPs) is transforming growth factor (TGF)-β1. The goal of this study was to de...
Embryo survival in the oviduct not significantly influenced by major histocompatibility complex social signaling in the horse.
Scientific reports    January 23, 2020   Volume 10, Issue 1 1056 doi: 10.1038/s41598-020-58056-w
Jeannerat E, Marti E, Thomas S, Herrera C, Sieme H, Wedekind C, Burger D.The major histocompatibility complex (MHC) influences sexual selection in various vertebrates. Recently, MHC-linked social signaling was also shown to influence female fertility in horses (Equus caballus) diagnosed 17 days after fertilization. However, it remained unclear at which stage the pregnancy was terminated. Here we test if MHC-linked cryptic female choice in horses happens during the first days of pregnancy, i.e., until shortly after embryonic entrance into the uterus and before fixation in the endometrium. We exposed estrous mares to one of several unrelated stallions, instrumentally...
A Common Practice of Widespread Antimicrobial Use in Horse Production Promotes Multi-Drug Resistance.
Scientific reports    January 22, 2020   Volume 10, Issue 1 911 doi: 10.1038/s41598-020-57479-9
Álvarez-Narváez S, Berghaus LJ, Morris ERA, Willingham-Lane JM, Slovis NM, Giguere S, Cohen ND.The practice of prophylactic administration of a macrolide antimicrobial with rifampin (MaR) to apparently healthy foals with pulmonary lesions identified by thoracic ultrasonography (i.e., subclinically pneumonic foals) is common in the United States. The practice has been associated epidemiologically with emergence of R. equi resistant to MaR. Here, we report direct evidence of multi-drug resistance among foals treated with MaR. In silico and in vitro analysis of the fecal microbiome and resistome of 38 subclinically pneumonic foals treated with either MaR (n = 19) or gallium maltolate (...
Genomic inbreeding trends, influential sire lines and selection in the global Thoroughbred horse population.
Scientific reports    January 16, 2020   Volume 10, Issue 1 466 doi: 10.1038/s41598-019-57389-5
McGivney BA, Han H, Corduff LR, Katz LM, Tozaki T, MacHugh DE, Hill EW.The Thoroughbred horse is a highly valued domestic animal population under strong selection for athletic phenotypes. Here we present a high resolution genomics-based analysis of inbreeding in the population that may form the basis for evidence-based discussion amid concerns in the breeding industry over the increasing use of small numbers of popular sire lines, which may accelerate a loss of genetic diversity. In the most comprehensive globally representative sample of Thoroughbreds to-date (n = 10,118), including prominent stallions (n = 305) from the major bloodstock regions of the w...
The composition of the perinatal intestinal microbiota in horse.
Scientific reports    January 16, 2020   Volume 10, Issue 1 441 doi: 10.1038/s41598-019-57003-8
Husso A, Jalanka J, Alipour MJ, Huhti P, Kareskoski M, Pessa-Morikawa T, Iivanainen A, Niku M.The establishment of the intestinal microbiota is critical for the digestive and immune systems. We studied the early development of the rectal microbiota in horse, a hindgut fermenter, from birth until 7 days of age, by qPCR and 16S rRNA gene amplicon sequencing. To evaluate initial sources of the foal microbiota, we characterised dam fecal, vaginal and oral microbiotas. We utilised an amplicon sequence variant (ASV) pipeline to maximise resolution and reproducibility. Stringent ASV filtering based on prevalence and abundance in samples and controls purged contaminants while preserving intest...
The neurocognitive gains of diagnostic reasoning training using simulated interactive veterinary cases.
Scientific reports    December 27, 2019   Volume 9, Issue 1 19878 doi: 10.1038/s41598-019-56404-z
Nassar M.The present longitudinal study ascertained training-associated transformations in the neural underpinnings of diagnostic reasoning, using a simulation game named "Equine Virtual Farm" (EVF). Twenty participants underwent structural, EVF/task-based and resting-state MRI and diffusion tensor imaging (DTI) before and after completing their training on diagnosing simulated veterinary cases. Comparing playing veterinarian versus seeing a colorful image across training sessions revealed the transition of brain activity from scientific creativity regions pre-training (left middle frontal and temporal...
High-starch diets alter equine faecal microbiota and increase behavioural reactivity.
Scientific reports    December 9, 2019   Volume 9, Issue 1 18621 doi: 10.1038/s41598-019-54039-8
Bulmer LS, Murray JA, Burns NM, Garber A, Wemelsfelder F, McEwan NR, Hastie PM.Gut microbiota have been associated with health, disease and behaviour in several species and are an important link in gut-brain axis communication. Diet plays a key role in affecting the composition of gut microbiota. In horses, high-starch diets alter the hindgut microbiota. High-starch diets are also associated with increased behavioural reactivity in horses. These changes in microbiota and behaviour may be associated. This study compares the faecal microbiota and behaviour of 10 naïve ponies. A cross-over design was used with experimental groups fed high-starch (HS) or high-fibre (HF) die...
Application of extracellular flux analysis for determining mitochondrial function in mammalian oocytes and early embryos.
Scientific reports    November 14, 2019   Volume 9, Issue 1 16778 doi: 10.1038/s41598-019-53066-9
Muller B, Lewis N, Adeniyi T, Leese HJ, Brison DR, Sturmey RG.Mitochondria provide the major source of ATP for mammalian oocyte maturation and early embryo development. Oxygen Consumption Rate (OCR) is an established measure of mitochondrial function. OCR by mammalian oocytes and embryos has generally been restricted to overall uptake and detailed understanding of the components of OCR dedicated to specific molecular events remains lacking. Here, extracellular flux analysis (EFA) was applied to small groups of bovine, equine, mouse and human oocytes and bovine early embryos to measure OCR and its components. Using EFA, we report the changes in mitochondr...
Comprehensive genome and transcriptome analyses reveal genetic relationship, selection signature, and transcriptome landscape of small-sized Korean native Jeju horse.
Scientific reports    November 13, 2019   Volume 9, Issue 1 16672 doi: 10.1038/s41598-019-53102-8
Srikanth K, Kim NY, Park W, Kim JM, Kim KD, Lee KT, Son JH, Chai HH, Choi JW, Jang GW, Kim H, Ryu YC, Nam JW, Park JE, Kim JM, Lim D.The Jeju horse, indigenous to the Jeju Island in Korea may have originated from Mongolian horses. Adaptations to the local harsh environment have conferred Jeju horse with unique traits such as small-sized body, stocky head, and shorter limbs. These characteristics have not been studied previously at the genomic level. Therefore, we sequenced and compared the genome of 41 horses belonging to 6 breeds. We identified numerous breed-specific non-synonymous SNPs and loss-of-function mutants. Demographic and admixture analyses showed that, though Jeju horse is genetically the closest to the Mongol...
CENP-A binding domains and recombination patterns in horse spermatocytes.
Scientific reports    November 1, 2019   Volume 9, Issue 1 15800 doi: 10.1038/s41598-019-52153-1
Centromeres exert an inhibitory effect on meiotic recombination, but the possible contribution of satellite DNA to this "centromere effect" is under debate. In the horse, satellite DNA is present at all centromeres with the exception of the one from chromosome 11. This organization of centromeres allowed us to investigate the role of satellite DNA on recombination suppression in horse spermatocytes at the stage of pachytene. To this aim we analysed the distribution of the MLH1 protein, marker of recombination foci, relative to CENP-A, marker of centromeric function. We demonstrated that the sa...
Antigen array for serological diagnosis and novel allergen identification in severe equine asthma.
Scientific reports    October 23, 2019   Volume 9, Issue 1 15170 doi: 10.1038/s41598-019-51820-7
White SJ, Moore-Colyer M, Marti E, Hannant D, Gerber V, Coüetil L, Richard EA, Alcocer M.Severe equine asthma (sEA), which closely resembles human asthma, is a debilitating and performance-limiting allergic respiratory disorder which affects 14% of horses in the Northern Hemisphere and is associated with increased allergen-specific immunoglobulin E (IgE) against a range of environmental proteins. A comprehensive microarray platform was developed to enable the simultaneous detection of allergen-specific equine IgE in serum against a wide range of putative allergenic proteins. The microarray revealed a plethora of novel pollen, bacteria, mould and arthropod proteins significant in t...
Species-specific and collection method-dependent differences in endometrial susceptibility to seminal plasma-induced RNA degradation.
Scientific reports    October 21, 2019   Volume 9, Issue 1 15072 doi: 10.1038/s41598-019-51413-4
Fernandez-Fuertes B, Sánchez JM, Bagés-Arnal S, McDonald M, Yeste M, Lonergan P.This study aimed to determine the effect of bull seminal plasma (SP) and sperm on endometrial function. Bovine endometrial explants were incubated with: ejaculated sperm with or without SP, epididymal sperm, or SP alone. Neither ejaculated nor epididymal sperm induced differential expression of IL1A, IL1B, IL6, IL8, PTGES2, TNFA, and LIF. Interestingly, SP had a detrimental effect on endometrial RNA integrity. Addition of an RNase inactivation reagent to SP blocked this effect, evidencing a role for a SP-RNase. Because bulls deposit the ejaculate in the vagina, we hypothesized that the bovine ...
Development of the equine gut microbiota.
Scientific reports    October 8, 2019   Volume 9, Issue 1 14427 doi: 10.1038/s41598-019-50563-9
Lindenberg F, Krych L, Kot W, Fielden J, Frøkiær H, van Galen G, Nielsen DS, Hansen AK.Shortly after birth the mammalian gut is colonized, by a transient microbiota, highly susceptible to environment and diet, that eventually stabilizes and becomes the resident gut microbiota. In a window of opportunity during the colonization, oral tolerance is established towards resident bacteria. In this study, the development of the equine gut microbiota was investigated in ten foals from parturition until post weaning. We found great differences in the core species of the gut microbiota composition between time-matched samples on Day 7 and 20 post-partum. Between day 20 and Day 50 post-par...
Expression of immune regulatory genes correlate with the abundance of specific Clostridiales and Verrucomicrobia species in the equine ileum and cecum.
Scientific reports    September 3, 2019   Volume 9, Issue 1 12674 doi: 10.1038/s41598-019-49081-5
Lindenberg F, Krych L, Fielden J, Kot W, Frøkiær H, van Galen G, Nielsen DS, Hansen AK.Billions of bacteria inhabit the gastrointestinal tract. Immune-microbial cross talk is responsible for immunological homeostasis, and symbiotic microbial species induce regulatory immunity, which helps to control the inflammation levels. In this study we aimed to identify species within the equine intestinal microbiota with the potential to induce regulatory immunity. These could be future targets for preventing or treating low-grade chronic inflammation occurring as a result of intestinal microbial changes and disruption of the homeostasis. 16S rRNA gene amplicon sequencing was performed on ...
Horses associate individual human voices with the valence of past interactions: a behavioural and electrophysiological study.
Scientific reports    August 9, 2019   Volume 9, Issue 1 11568 doi: 10.1038/s41598-019-47960-5
Brain lateralization is a phenomenon widely reported in the animal kingdom and sensory laterality has been shown to be an indicator of the appraisal of the stimulus valence by an individual. This can prove a useful tool to investigate how animals perceive intra- or hetero-specific signals. The human-animal relationship provides an interesting framework for testing the impact of the valence of interactions on emotional memories. In the present study, we tested whether horses could associate individual human voices with past positive or negative experiences. Both behavioural and electroencephalo...