Stranger danger? An investigation into the influence of human-horse bond on stress and behaviour

Human-animal bond is receiving increasing attention and is thought to confer benefits on well-being and performance in working animals. One important benefit of bonding is the "safe base" an attachment figure provides, which manifests in better coping and increased exploration during potential threat. However, there is limited research exploring the existence or benefits of human-horse bonds, though bonding is sought after by both pleasure and elite riders. The purpose of the current study was to determine whether the presence of horses' owners confers a safe-base, therefore improving horse behaviour and physiological stress responses during novel handling tests. Horses completed two different handling tests, one with their owner and the other with an unfamiliar experimental handler (n = 46). Test and handler order was randomised and handlers were double blind to the performance of the horse with the alternate handler. Time taken to complete the tests and proactive behaviour were measured as indicators of performance and compliance. Core temperature, discrepancy in eye temperature, heart rate and heart rate variability were recorded to assess stress responses. If horses experience a "safe base" effect in the vicinity of their owner, they would be expected to show lower stress responses and greater behavioural compliance, compared to being handled by a stranger. There was no difference in behaviour or any physiological stress response between the handlers. This indicates that a calm, competent, but unknown handler may be equally effective to an owner during stressful procedures as neither equine performance nor affective state supported a safe-base effect. This supports previous research suggesting that the level of bond between human and horse may not be the most salient factor in coping or compliance during training and handling. These findings have implications for veterinary and clinical behaviour counselling, where novel human handlers must modify behaviour under potentially stressful circumstances

Poker face: discrepancies in behaviour and affective states in horses during stressful handling procedures

Correct assessment of stress in horses is important for both horse welfare and handler safety during necessary aversive procedures. Handlers depend on behaviour when judging how well an individual is tolerating stressful procedures such as loading or veterinary intervention. However, evidence suggests that behaviour may not accurately reflect affective states in horses. This may be explained by individual differences in coping styles, which have tentatively been identified in horses. The current study assessed whether behaviour during two novel handling procedures was associated with physiological indicators of stress. Core temperature, discrepancy in eye temperature and heart rate variability (HRV) were compared with compliance and proactivity shown by horses during two novel handling tests (n = 46). Test A required subjects to cross a large blue tarpaulin on the ground. Test B required subjects to walk through plastic streamers suspended overhead. Physiological indicators of stress did not correlate with time taken to complete the handling tests. This indicates some subjects crossed an object they found aversive. Crossing time may be influenced more by stimulus-control than the level of aversion experienced. The level of proactivity shown was not associated with HRV, HR, core temperature or the discrepancy in temperature between eyes. This suggests that proactive horses, which appear more stressed, show similar stress responses to more reactive individuals. These findings support previous research indicating that behaviour commonly used within the equestrian industry may not provide reliable indicators of a horse’s ability to tolerate a stressful procedure. The influence of training and the extent to which a horse is under stimulus-control may over-shadow inherent emotional responses, with implications for handler safety and horse welfare

Dually noted: the effects of a pressure headcollar on compliance, discomfort and stress in horses during handling

Horse handlers often encounter problem behaviour resulting from a lack of stimulus control. Handlers are often only 15% of the weight of horses, which evolved strong flight responses. Therefore, many riders and handlers resort to the use of "aids" to maintain control of their animals. However, there are increasing concerns about the efficacy and welfare implication of such devices, particularly when applied to sensitive facial structures. One such device is a Dually® headcollar which aims to increase compliance. Despite its popularity, little is known about the effects of this aid on behaviour or stress. The aim of the current study was to determine whether the use of a Dually headcollar improves compliance during handling and, if so, whether this might be achieved with concomitant increases in stress or discomfort. Subjects completed two novel handling tests, one wearing a Dually with a line attached to the pressure mechanism and one attached to the standard ring as a Control. Crossing time and proactive behaviour were recorded as indicators of compliance. Core temperature and the discrepancy between eye temperatures were measured using IRT before and after testing as an indicator of stress. The Horse Grimace Scale (HGS) was used to measure discomfort caused by each configuration of the device. The Dually did not result in more compliant behaviour, compared to the Control (p=0.935; p=0.538). However, the Dually configuration did result in a significantly higher HGS scores (p=0.034). This may indicate that there is an impact on animal welfare by using this device that is not justified by improved behaviour. However, IRT readings of core temperature (p=0.186) and discrepancy between the eyes (p=0.972) did not indicate the Dually increased stress in subjects. Taken together, this suggests the Dually is ineffective in naïve horses but causes increased discomfort

Zylkéne to load? The effects of alpha-casozepine on compliance and coping in horses during loading

Horses are routinely transported for access to safe off-road riding, veterinary care, breeding, sale or moving to a new home environment. However, transport is a known stressor in horses. For this reason, problem behavior when loading is a commonly reported issue which presents risks to handlers and horse welfare. Existing literature and manufacturers recommendations suggests that alpha-casozepine may be effective in improving the behavior and welfare of horses during loading onto a vehicle for transport. The current paper aims to assess the behavioral and physiological effects of a commercially available alpha-casozepine feed supplement (Zylkéne Equine) in horses during loading and confinement on a transport lorry. Subjects (n = 10) were loaded once with the supplement and once without, in a balanced random order with each subject acted as their own control. The handler was blind to treatment. Time to load onto the lorry, and movement of feet, licking and chewing, and vocalising within the lorry, were recorded as behavioral indicators of compliance and coping. Heart rate, heart rate variability, salivary cortisol, and infrared thermography of both core temperature and the discrepancy between eyes, were measured as indicators of arousal. There were no significant differences in physiology between Treatment and Control (P > 0.05). Treatment resulted in a significantly shorter Loading Time than control (P = 0.04), however, the actual difference in median time was only 0.45 seconds. No other behavioral indicator differed between Treatment and Control (P > 0.05). Power analysis revealed the sample was sufficient to detect a significant effect. Where modest effects were observed for a small number of variables, Treatment effect contradicted predictions. Taken together, this indicates that alpha-casozepine does not affect a horse’s ability to cope with loading and confinement in a horse lorry. Further work is required to ascertain whether the maximum dosage – twice that used here – might affect coping and behavior in horses

Wolbachia in the flesh: symbiont intensities in germ-line and somatic tissues challenge the conventional view of Wolbachia transmission routes

Symbionts can substantially affect the evolution and ecology of their hosts. The investigation of the tissue-specific distribution of symbionts (tissue tropism) can provide important insight into host-symbiont interactions. Among other things, it can help to discern the importance of specific transmission routes and potential phenotypic effects. The intracellular bacterial symbiont Wolbachia has been described as the greatest ever panzootic, due to the wide array of arthropods that it infects. Being primarily vertically transmitted, it is expected that the transmission of Wolbachia would be enhanced by focusing infection in the reproductive tissues. In social insect hosts, this tropism would logically extend to reproductive rather than sterile castes, since the latter constitute a dead-end for vertically transmission. Here, we show that Wolbachia are not focused on reproductive tissues of eusocial insects, and that non-reproductive tissues of queens and workers of the ant Acromyrmex echinatior, harbour substantial infections. In particular, the comparatively high intensities of Wolbachia in the haemolymph, fat body, and faeces, suggest potential for horizontal transmission via parasitoids and the faecal-oral route, or a role for Wolbachia modulating the immune response of this host. It may be that somatic tissues and castes are not the evolutionary dead-end for Wolbachia that is commonly thought

Dually investigated: the effect of a pressure headcollar on the behaviour, discomfort and stress of trained horses

The Dually™ is a control headcollar designed to improve equine behaviour during handling challenges by applying greater pressure than a standard headcollar. Previous research indicated it did not improve compliance in naïve horses but did result in higher Horse Grimace Scale scores (HGS) indicative of discomfort. However, subjects had not been trained to step forward to release the pressure applied by the headcollar. The current study aimed to determine the effect of training on behaviour and physiology of horses wearing the Dually™ headcollar during handling challenges. To this end, subjects received three training sessions prior to completing two handling tests in which they crossed distinct novel obstacles, one wearing a Dually™ with a line attached to the pressure mechanism and one attached to the standard ring as a control. Behaviour was coded by hypothesis blind researchers: time to cross the obstacle and proactive refusal (moving away from the obstacle) were recorded as indicators of compliance and the Horse Grimace Scale was used to measure discomfort caused by each configuration of the device. Infrared thermography of ocular temperature, heart rate variability (RMSSD and low/high frequency ratios (LF/HF)) and salivary cortisol were measured as indicators of arousal. Data from the previous study on Naïve horses was also included to compare responses to the Dually in Naïve and Trained horses. Training resulted in a decrease in RMSSD (p = 0.002) and an increase in LF/HF (p=0.012), compared to rest, indicating arousal. As per the original study, horses did not complete the tests more quickly in the Dually, compared to control (p=0.698). Trained horses from this study tended to be more proactive in the Dually compared to Controls (p=0.066) and significantly more so than Naïve horses from the previous study (p=0.002) suggesting that behaviour becomes less desirable during early Dually training. Yet, stress and HGS indicators were not higher in the Dually compared to Control during testing. Results suggest the Dually has a negative effect on behaviour but not on stress or discomfort during short handling challenges. Further research is warranted to determine the long-term effect of Dually experience on behaviour and welfare

Inactivation of TIF1γ Cooperates with KrasG12D to Induce Cystic Tumors of the Pancreas

Inactivation of the Transforming Growth Factor Beta (TGFβ) tumor suppressor pathway contributes to the progression of Pancreatic Ductal AdenoCarcinoma (PDAC) since it is inactivated in virtually all cases of this malignancy. Genetic lesions inactivating this pathway contribute to pancreatic tumor progression in mouse models. Transcriptional Intermediary Factor 1 gamma (TIF1γ) has recently been proposed to be involved in TGFβ signaling, functioning as either a positive or negative regulator of the pathway. Here, we addressed the role of TIF1γ in pancreatic carcinogenesis. Using conditional Tif1γ knockout mice (Tif1γlox/lox), we selectively abrogated Tif1γ expression in the pancreas of Pdx1-Cre;Tif1γlox/lox mice. We also generated Pdx1-Cre;LSL-KrasG12D;Tif1γlox/lox mice to address the effect of Tif1γ loss-of-function in precancerous lesions induced by oncogenic KrasG12D. Finally, we analyzed TIF1γ expression in human pancreatic tumors. In our mouse model, we showed that Tif1γ was dispensable for normal pancreatic development but cooperated with Kras activation to induce pancreatic tumors reminiscent of human Intraductal Papillary Mucinous Neoplasms (IPMNs). Interestingly, these cystic lesions resemble those observed in Pdx1-Cre;LSL-KrasG12D;Smad4lox/lox mice described by others. However, distinctive characteristics, such as the systematic presence of endocrine pseudo-islets within the papillary projections, suggest that SMAD4 and TIF1γ don't have strictly redundant functions. Finally, we report that TIF1γ expression is markedly down-regulated in human pancreatic tumors by quantitative RT–PCR and immunohistochemistry supporting the relevance of these findings to human malignancy. This study suggests that TIF1γ is critical for tumor suppression in the pancreas, brings new insight into the genetics of pancreatic cancer, and constitutes a promising model to decipher the respective roles of SMAD4 and TIF1γ in the multifaceted functions of TGFβ in carcinogenesis and development

The Anti-Tumor Effect of HDAC Inhibition in a Human Pancreas Cancer Model Is Significantly Improved by the Simultaneous Inhibition of Cyclooxygenase 2

Pancreatic ductal adenocarcinoma is the fourth leading cause of cancer death worldwide, with no satisfactory treatment to date. In this study, we tested whether the combined inhibition of cyclooxygenase-2 (COX-2) and class I histone deacetylase (HDAC) may results in a better control of pancreatic ductal adenocarcinoma. The impact of the concomitant HDAC and COX-2 inhibition on cell growth, apoptosis and cell cycle was assessed first in vitro on human pancreas BxPC-3, PANC-1 or CFPAC-1 cells treated with chemical inhibitors (SAHA, MS-275 and celecoxib) or HDAC1/2/3/7 siRNA. To test the potential antitumoral activity of this combination in vivo, we have developed and characterized, a refined chick chorioallantoic membrane tumor model that histologically and proteomically mimics human pancreatic ductal adenocarcinoma. The combination of HDAC1/3 and COX-2 inhibition significantly impaired proliferation of BxPC-3 cells in vitro and stalled entirely the BxPC-3 cells tumor growth onto the chorioallantoic membrane in vivo. The combination was more effective than either drug used alone. Consistently, we showed that both HDAC1 and HDAC3 inhibition induced the expression of COX-2 via the NF-kB pathway. Our data demonstrate, for the first time in a Pancreatic Ductal Adenocarcinoma (PDAC) model, a significant action of HDAC and COX-2 inhibitors on cancer cell growth, which sets the basis for the development of potentially effective new combinatory therapies for pancreatic ductal adenocarcinoma patients.Peer reviewe