Applied Animal Behaviour Science最新文献

{"title":"Keel bone fractures are associated with anxiety like behavioural responses in laying hens housed individually in furnished cages","authors":"Jiaqi Zhang ,&nbsp;Jingyi Zhang ,&nbsp;Haoran Zhang ,&nbsp;Lu Guo ,&nbsp;Hengyi Zhang ,&nbsp;Hongui Liu ,&nbsp;Jianhong Li ,&nbsp;Jun Bao ,&nbsp;Runxiang Zhang","doi":"10.1016/j.applanim.2025.106903","DOIUrl":"10.1016/j.applanim.2025.106903","url":null,"abstract":"<div><div>Keel bone fracture (KBF) is a common injury that affects the behaviour and welfare of laying hens. This study aimed to investigate the impact of KBF on anxiety-like behaviour in laying hens and explore the underlying regulatory mechanisms. Ninety 46-week-old Hy-Line Brown hens were individually housed in furnished cages and divided into two groups based on keel bone diagnosis: the non-fracture group (NF) and the KBF group. Keel bone status was evaluated at 46, 49, and 52 weeks of age using a combination of palpation and X-ray examination. Behavioural data were collected two days after each diagnosis. The open field test (OFT) was conducted at 49 weeks (n = 25 per group), and the attention bias test (ABT) was performed at 52 weeks (NF = 39, KBF=51). Behavioural observations revealed that hens with KBF displayed significantly increased preening and toe pecking behaviours. OFT showed a significant prolongation of the first movement latency and a reduction in exploration time in KBF hens. KBF hens exhibited significantly longer latencies for the first vocalization, first step, and first eating in ABT. Collectively, the OFT and ABT results indicated that hens with keel bone fractures exhibited the anxiety-like behaviour. Physiological indicators (n = 6) revealed elevated serum corticosterone levels and decreased serum serotonin levels in hens with KBF, suggesting that KBF induces a stress response, which contributes to the manifestation of anxiety-like behaviours. Golgi staining (n = 3) revealed a significant reduction in hippocampal neuronal complexity in KBF hens, indicating neuronal damage. qRT-PCR (n = 6) and western blot (n = 3) analysis demonstrated altered expression of genes related to anxiety in the hippocampus, characterized by a notable decrease in the protein levels of DCX, BDNF, and EGR1, suggesting impaired neurogenesis (DCX), reduced stress resilience (DCX and BDNF), and decrease neuroplasticity (BDNF and EGR1), all of which are closely linked to anxiety-like behaviour. Therefore, we suggest that keel bone fracture is associated with anxiety-like behaviour in laying hens, which may result from stress signals generated by the fracture that trigger the expression of anxiety-related genes in the brain.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"296 ","pages":"Article 106903"},"PeriodicalIF":2.0,"publicationDate":"2025-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Excessive energy in dogs: The impact of caregiver-related variables and fear/anxiety","authors":"Maria Toscano Batista ,&nbsp;Catarina Lavrador ,&nbsp;Gonçalo da Graça-Pereira","doi":"10.1016/j.applanim.2025.106900","DOIUrl":"10.1016/j.applanim.2025.106900","url":null,"abstract":"<div><div>Excessive energy (EE) and hyperactivity are frequent behavioral problems in dogs, with significant implications for canine welfare and caregiver well-being, yet they remain less studied than reactivity/aggression or separation-related problems. This study examined the influence of caregiver-related variables on EE in dogs, as well as the role of fear/anxiety. Data were collected through an online questionnaire from 730 caregivers, and multiple linear regression analyses were conducted. Younger dogs showed higher levels of EE. Among caregiver-related variables, stress, higher perceived caregiving costs, and more positive attitudes towards aversive training emerged as risk factors. In contrast, the ability to recognize chronic pain, knowledge of basic behaviors, and medication use were protective. The final regression model explained 21.3 % of the variance, with dog age, caregiver stress, and attitudes towards aversive training being the strongest predictors. When fear/anxiety was added, the explained variance rose to 29.5 %, underscoring its relevance as an emotional comorbidity. These findings highlight the central role of caregiver mental health, perceptions, and training practices in shaping EE, while also emphasizing the protective value of pain recognition, education, and medical support. Interventions should therefore integrate caregiver well-being, training strategies, pain assessment, and dogs’ emotional states, in line with a One Welfare perspective.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"296 ","pages":"Article 106900"},"PeriodicalIF":2.0,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effects of cannabidiol (CBD) in transport water on the behaviour of ornamental fishes","authors":"M. Jones ,&nbsp;M.E. Alexander ,&nbsp;M. Vanderzwalmen ,&nbsp;D. Snellgrove ,&nbsp;M. Talbot ,&nbsp;P. Smith ,&nbsp;F.L. Henriquez ,&nbsp;I. McLellan ,&nbsp;K.A. Sloman","doi":"10.1016/j.applanim.2025.106897","DOIUrl":"10.1016/j.applanim.2025.106897","url":null,"abstract":"<div><div>The ornamental fish supply chain has multiple transportation phases which can induce stress in fishes. Previous studies have considered methods of improving welfare during transport, by adding water conditioners based on natural compounds known to have anxiolytic effects. Cannabidiol (CBD) has recently emerged as a compound of interest with beneficial immunomodulatory, anti-inflammatory, and anxiolytic effects in mammals. In the first part of the present study, we identified whether addition of CBD to the transport water of ornamental fishes (at nominal concentrations of 3.9, 7.8 or 15.6 mg/l) had an effect on group behaviour post-transport. Variatus platys were transported for 30 min in bags containing one of five treatment groups (control, solvent control, 3.9, 7.8 or 15.6 mg/l CBD). They were then videoed as a group (15 min) immediately after introduction post-transport into an empty tank, with further videoing performed 30 min and 2 h after release. Behaviours analysed included biting, chasing, erratic movements and time spent immobile. The lowest concentration found to affect a range of behaviours was 7.8 mg/l (the middle concentration). Based on these findings, this concentration was used in a follow-on study to identify whether the use of CBD during transport affected individual behaviour and physiology post-transport. Fish were transported in the same way, and then fish were placed individually into open field arenas immediately after transport and videoed for 15 min. Behaviours analysed in the open field arenas included distance travelled, mean speed, time spent immobile, and time spent in the central zone. Water cortisol and skin mucus quantity were also analysed. CBD significantly affected behaviour post-transport, with those fish exposed to CBD exhibiting significantly reduced stress-related behaviours than those in the control and solvent control groups at both the group and individual level. No effects on mucus or water cortisol were seen. These findings highlight the potential for using CBD within commercial water conditioners to reduce the effects of transport stress for ornamental fishes.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"296 ","pages":"Article 106897"},"PeriodicalIF":2.0,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145838164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Behavior-related changes in canine heart rate and heart rate variability during short-term measurement","authors":"Päivi Berg ,&nbsp;Aija Koskela ,&nbsp;Jukka A. Lipponen ,&nbsp;Heini Törnqvist ,&nbsp;Jan Kujala ,&nbsp;Mika P. Tarvainen ,&nbsp;Miiamaaria V. Kujala","doi":"10.1016/j.applanim.2025.106899","DOIUrl":"10.1016/j.applanim.2025.106899","url":null,"abstract":"<div><div>Cardiac function is influenced by both physiological and emotional factors, with heart rate variability (HRV) serving as a key indicator of health and wellbeing. Despite the growing interest in utilizing short-term cardiac measures in canine science, the behavioral aspects of HRV in domestic dogs (<em>Canis familiaris</em>) remain largely unexplored. This study aimed to provide reference values for short-term HRV to aid future research on canine behavior in medium-sized, mesocephalic and dolichocephalic dogs, to examine differences in HRV during typical canine behaviors, and to develop practical tools for the analysis of canine cardiac function. We assessed heart rate, HRV, and physical activity of 29 dogs across five behavioral states (Resting, Playing, Panting, Spontaneous sniffing and Food searching) and investigated how these behaviors influenced time-domain and frequency-domain HRV parameters. The impact of physical activity, sex, neutered status, age, height, and weight on these parameters within the specific sample was also assessed. Both time-domain and frequency-domain parameters were affected by the behaviors. Precisely, HRV generally decreased with behavior-related physical activity (root mean square of successive differences, RMSSD; Resting vs. Playing, p &lt; 0.001; and Resting vs. Searching for food, p &lt; 0.001). However, RMSSD was significantly lower during Searching for food compared to Spontaneous sniffing (p = 0.012), despite similar activity levels, indicating higher emotional arousal when searching for food. Overall, the high-frequency component (HF power) and RMSSD differentiated well between the distinct canine behaviors. Also, physical activity (measured as 3D acceleration) was the most influential background variable in this highly specific sample, correlating with HRV parameters and depending on the behavior.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"296 ","pages":"Article 106899"},"PeriodicalIF":2.0,"publicationDate":"2025-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145749958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Use of artificial shelter structures by individual Penaeus monodon broodstock prawns","authors":"Dana L.M. Campbell ,&nbsp;Greg J. Coman ,&nbsp;Chris Stratford ,&nbsp;Pushkar Mandrekar ,&nbsp;Troy Kalinowski ,&nbsp;Caroline Lee ,&nbsp;Artur N. Rombenso","doi":"10.1016/j.applanim.2025.106896","DOIUrl":"10.1016/j.applanim.2025.106896","url":null,"abstract":"<div><div>Optimising the housing and management conditions for aquaculture species is becoming increasingly important. Determining what resources animals prefer can inform captive management decisions for positive impacts on the individual’s welfare. The black tiger prawn (<em>Penaeus monodon</em>) aquaculture industry is prominent in Australia and is the second most farmed prawn species internationally, however there is little information on resource preferences of broodstock animals in hatcheries. The objective of this study was to assess the use of three different shelter structures, including a rectangular box, a cylindrical tube, and a vertical piece of plastic mesh, by 21 individual male <em>P. monodon</em>. It was hypothesised that prawns would utilise these objects and would prefer to be hidden rather than out on the open sandy substrate. Observations were made from video-recordings of individual prawns in separate tanks across 72 h. Scan sampling every 15 min included the location of the prawn in the tank (visually divided into thirds), proximity to a shelter structure, and contact with a shelter structure. Analyses across 12-h time blocks showed no effect of time on location (both <em>P</em> ≥ 0.13). There were only 6 instances where the prawns were observed in contact with the shelter structures and no significant difference in proximity preferences for the three shelter structures (<em>P</em> = 0.26). Large standard deviations across measures of location preference and proximity indicated high variability between the tested individuals. Contrary to predictions, this study indicates these selected objects did not provide sufficient value to individual prawns to facilitate engagement with them in the experimental context tested. It is possible the sandy substrate in the tanks was sufficient enrichment for the prawns or that use may have increased if there was conspecific competition for access. Further work should continue to explore what resources may be utilised by this species that may inform on practical additions in the commercial hatchery environment. Future identification of resources that are of value to <em>P. monodon</em> prawns should continue to assess preferences while taking into account communal settings, individual variation and the potential benefits of providing a choice of resources that can cater to differing preferences.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"296 ","pages":"Article 106896"},"PeriodicalIF":2.0,"publicationDate":"2025-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Is social buffering consistent across the time of the day? A study in a diurnal (Nile tilapia Oreochromis niloticus) and a nocturnal fish species (tench Tinca tinca)","authors":"Santiago Pintos ,&nbsp;Tyrone Lucon-Xiccato ,&nbsp;Gonzalo De Alba ,&nbsp;Francelly Geralda Campos ,&nbsp;Francisco Javier Sánchez-Vázquez ,&nbsp;Cristiano Bertolucci ,&nbsp;Luisa María Vera","doi":"10.1016/j.applanim.2025.106895","DOIUrl":"10.1016/j.applanim.2025.106895","url":null,"abstract":"<div><div>In social animals, interactions with conspecifics provide several benefits that ultimately enhance welfare and fitness, including the attenuation of individuals’ responses to environmental challenges often referred to as social buffering. This effect has been reported across several fish taxa. However, evidence suggests that social buffering is remarkably variable in fish, depending on intrinsic factors of species and environmental conditions. In this research, we explored whether social buffering effects vary throughout the time of the day in two fish species with opposed activity patterns: the diurnal Nile tilapia (<em>Oreochromis niloticus)</em> and the nocturnal tench (<em>Tinca tinca)</em>. To this goal, we collected behavioural indicators of welfare in fish subjected to two different social conditions (i.e., isolation or groups of five fish) for 24 h every 4 h. These indicators were based on well-reported behaviours associated with stress in fish, such as bottom-dwelling, freezing, activity and erratic movements. Our results revealed social buffering effects in both species in almost all analysed indicators, showing increased welfare in the presence of conspecifics. Critically, while the social buffering effect was constant throughout the day in tench, it varied between day and night in tilapia. Furthermore, the isolation condition disrupted daily rhythmicity only in tilapia, but not in tench, highlighting interspecific variation in the influence of conspecifics on behavioural rhythms. Overall, our findings supported the presence of social companions to improve fish welfare in captivity and underscored the importance of considering species-specific behavioural rhythms for tailoring management practices that minimise stress.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"296 ","pages":"Article 106895"},"PeriodicalIF":2.0,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145749957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Born to lunge? A survey on lunging behaviour and associated factors in Border Collies","authors":"Britt Marin ,&nbsp;Sofie Salden ,&nbsp;Bart Broeckx ,&nbsp;Wim Van Den Broeck ,&nbsp;Anouck Haverbeke","doi":"10.1016/j.applanim.2025.106894","DOIUrl":"10.1016/j.applanim.2025.106894","url":null,"abstract":"<div><div>Lunging behaviour (LB) is defined as a sudden, forceful forward movement towards moving stimuli (e.g., cars, cyclists), typically resulting in abrupt leash tension and reflecting heightened arousal or motion sensitivity. It is commonly reported by Border Collie (BC) caregivers and is often perceived as problematic or dangerous, yet its underlying drivers remain poorly understood. Given that Border Collies (BCs) have been bred for herding—an activity involving high responsiveness to motion—this breed may be particularly predisposed. However, empirical data on the prevalence and influencing factors of lunging behaviour in BCs remains limited. This study therefore aimed to (1) determine the prevalence of LB in BCs, and (2) identify potential associations between LB and various demographic (i.e. age, lineage, source and age of adoption), behavioural (i.e. presence of abnormal repetitive behaviours (ARBs), and environmental (i.e. living environment, walking context, type of training) factors. An online survey was performed and resulted in 952 valid responses among BC caregivers. LB was reported in 25.3 % (n = 241) of BCs. Age, training type, and co-occurrence of abnormal repetitive behaviours (ARBs) were significantly associated with LB, while lineage and environment were not. These findings suggest that LB is multifactorial, possibly reflecting broader patterns of behavioural dysregulation with welfare implications. Greater awareness of the factors associated with LB may support more informed behavioural management strategies, ultimately improving the welfare of both these dogs and their caregivers.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"296 ","pages":"Article 106894"},"PeriodicalIF":2.0,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789199","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"King penguins and gentoo penguins prefer to behaviourally interact with white plastics over other colours","authors":"A.M. Heswall ,&nbsp;K.E. Cain ,&nbsp;M.R. Friesen ,&nbsp;A.C. Gaskett","doi":"10.1016/j.applanim.2025.106891","DOIUrl":"10.1016/j.applanim.2025.106891","url":null,"abstract":"<div><div>Seabirds are a speciose and at-risk animal group with specialized sensory ecologies that are highly responsive and vulnerable to sensory-related threats such as plastic ingestion. Seabirds may consume plastic because it functions as a sensory trap, exploiting the birds’ pre-existing biases for specific odours or colours. Previous studies have documented white plastics as a common colour in the ocean and is frequently ingested by seabirds, potentially due to its abundance. Alternatively, seabirds may selectively choose to ingest the colour white regardless of abundance in the marine environment. To evaluate whether seabirds prefer to behaviourally interact with white plastics over others, we used standardized behavioural assays to test seabird attraction to various colours of a commonly ingested plastic item: bottlecaps. Assays were performed with captive gentoo and king penguins (<em>Pygoscelis papua</em>; and <em>Aptenodytes patagonicus</em>; Tokoraki) by simultaneously presenting white, black, red and blue bottlecaps and quantifying responses. Overall, penguins were looking and showing interest with all four colours but strongly preferred the white bottlecap, showing a colour preference behaviour towards white plastics among seabirds. Seabirds may be actively attracted to white as it could be strongly associated with cues from eggs and prey associated with a potential foraging or reproductive benefit. It would be beneficial for future research to explore how plastic maybe exploiting the seabird’s sensory biases, using a sensory ecology approach to mitigate plastic ingestion for a threatened avian group.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"295 ","pages":"Article 106891"},"PeriodicalIF":2.0,"publicationDate":"2025-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145749009","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Behaviour and spatial distribution of rainbow trout (Oncorhynchus mykiss) in experimental tanks: Influence of water flow and husbandry procedures","authors":"Martina Bortoletti ,&nbsp;Miina Lõoke ,&nbsp;Francesco Pascoli ,&nbsp;Gaia Bacchin ,&nbsp;Andrea Meloni ,&nbsp;Anna Broseghini ,&nbsp;Daniela Bertotto ,&nbsp;Paolo Mongillo ,&nbsp;Lieta Marinelli","doi":"10.1016/j.applanim.2025.106892","DOIUrl":"10.1016/j.applanim.2025.106892","url":null,"abstract":"<div><div>Rainbow trout (<em>Oncorhynchus mykiss</em>) is increasingly employed as an experimental model, yet knowledge of its behaviour in laboratory settings is limited. This study examined short-term (24 h), mid-term (1 week), and circadian changes in behavioural and spatial patterns of individually-housed trout under two experimental conditions differing in the extent of underwater current: control <em>vs</em> water flow. Moreover, the effects of feeding and tank cleaning were evaluated to identify potential behavioural indicators of welfare of the species in this context. Short-term effects revealed that flow rate impacted on swimming behaviour, with trout showing less species-specific behavior in the absence of water flow. Over time, trout exhibited a marked reduction of behavioral expression, interpretable as behavioural stabilization but likely reflecting suboptimal welfare. Trout behaviors were affected by the day time being less active in mornings. Event-related findings indicated an increased in active behaviors and a decrease in maintenance behaviours after both husbandry procedures, with that of cleaning being more pronounced and lasting remarkably longer, suggesting that the two reactions were fundamentally different. This study highlights the need to critically evaluate behavioural expression, and the need to provide species-specific husbandry conditions. We also provide foundational insights into rainbow trout behaviour valuable for refining research practices and improving welfare assessment of this experimental fish.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"295 ","pages":"Article 106892"},"PeriodicalIF":2.0,"publicationDate":"2025-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145748959","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Behavioural and performance responses of primiparous buffalo cows to milking types with full or partial cow-calf contact during milking","authors":"İbrahim Cihangir Okuyucu","doi":"10.1016/j.applanim.2025.106893","DOIUrl":"10.1016/j.applanim.2025.106893","url":null,"abstract":"<div><div>The shift to mechanized milking warrants further investigation into its effects on performance and behavioral outcomes in buffalo (Bubalus bubalis), a species characterized by a unique mother-calf bond and udder structure. Therefore, this study aimed to evaluate the effects of milking type (MT) — bucket milking machine (BM) versus hand milking (HM) — and of cow–calf contact (CC) during milking (full (FC) or partial (PC)) on milking performance and behavioral traits in primiparous Anatolian buffalo cows. Hence, for an 8-week experiment, 80 healthy cows weighing 500 ± 40 kg and with 35 ± 5 days in milk were selected and assigned to four treatments (BM-FC, HM-FC, BM-PC, and HM-PC), with 20 cows per treatment. Data on behavioral traits (milking temperament, stepping, kicking, urinating, defecating, vocalization, turning the head back, and tail wagging) and milking performance (milk flow rate, milking duration, and milk yield per milking) were collected four times at 14-day intervals: behavioral traits by scoring and milking performance by measuring. The milking performance data were evaluated using a 2 × 2 factorial design: MT (BM and HM) × CC (PC and FC). A non-parametric permutation test was applied to assess the effects of these factors on the temperament score. Depending on the data type, Spearman or Pearson correlation analyses were used to assess pairwise correlations among behavioral and milking-performance traits. Binary logistic regression analysis was performed to examine the relationship between behavioral responses and the MT and CC factors. The BM resulted in a longer milking duration and a lower milk flow rate than the HM, whereas the FC resulted in a shorter milking duration and a higher milk flow rate than the PC. BM-PC showed reduced milk yield per milking and greater nervousness during milking compared with BM-FC, HM-PC, and HM-FC. Positive correlations between milking temperament score and behavioral traits ranged from weak (defecation and urination) to moderate (vocalizing, turning the head back, and tail wagging). Milking performance parameters and temperament scores were moderately negatively correlated. The BM and FC cows exhibited higher behavioral response levels (primarily kicking, vocalization, and stepping) than their reference counterparts (HM and PC). This study indicates that primiparous Anatolian buffalo cows subjected to BM are more susceptible to CC-associated milking stress, which can adversely affect their welfare and, subsequently, their milking performance.</div></div>","PeriodicalId":8222,"journal":{"name":"Applied Animal Behaviour Science","volume":"295 ","pages":"Article 106893"},"PeriodicalIF":2.0,"publicationDate":"2025-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145748961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}