A successful experimental model for intimal hyperplasia prevention using a resveratrol-delivering balloon.

OBJECTIVE: Restenosis due to intimal hyperplasia is a major clinical problem that compromises the success of angioplasty and endovascular surgery. Resveratrol (RSV) has demonstrated a beneficial effect on restenosis from angioplasty. Unfortunately, the physicochemical characteristics of RSV reduce the practicality of its immediate clinical application. This work proposes an experimental model aiming to setup an intravessel, elutable, RSV-containing compound. METHODS: A 140 µg/mL RSV sterile injectable solution with a suitable viscosity for intravascular administration by drug-delivery catheter (RSV-c) was prepared. This solution was locally administered in the common iliac artery of adult male New Zealand White rabbits using a dedicated device (Genie; Acrostak, Geneva, Switzerland) after the induction of intimal hyperplasia by traumatic angioplasty. The RSV concentrations in the wall artery were determined, and the thickness of the harvested iliac arteries was measured over a 1-month period. RESULTS: The Genie catheter was applied in rabbit vessels, and the local delivery resulted in an effective reduction in restenosis after plain angioplasty. Notably, RSV-c forced into the artery wall by balloon expansion might accumulate in the interstitial areas or within cells, avoiding the washout of solutions. Magnification micrographs showed intimal proliferation was significantly inhibited when RSV-c was applied. Moreover, no adverse events were documented in in vitro or in vivo studies. CONCLUSIONS: RSV can be advantageously administered in the arterial walls by a drug-delivery catheter to reduce the risk of restenosis.

Respiratory Electrodialysis. A Novel, Highly Efficient Extracorporeal CO2 Removal Technique.

RATIONALE: We developed an innovative, minimally invasive, highly efficient extracorporeal CO2 removal (ECCO2R) technique called respiratory electrodialysis (R-ED). OBJECTIVES: To evaluate the efficacy of R-ED in controlling ventilation compared with conventional ECCO2R technology. METHODS: Five mechanically ventilated swine were connected to a custom-made circuit optimized for R-ED, consisting of a hemofilter, a membrane lung, and an electrodialysis cell. Electrodialysis regionally modulates blood electrolyte concentration to convert bicarbonate to CO2 before entering the membrane lung, enhancing membrane lung CO2 extraction. All animals underwent three repeated experimental sequences, consisting of four steps: baseline (1 h), conventional ECCO2R (2 h), R-ED (2 h), and final NO-ECCO2R (1 h). Blood and gas flow were 250 ml/min and 10 L/min, respectively. Tidal volume was set at 8 ml/kg, and respiratory rate was adjusted to maintain arterial Pco2 at 50 mm Hg. MEASUREMENTS AND MAIN RESULTS: During R-ED, chloride and H(+) concentration increased in blood entering the membrane lung, almost doubling CO2 extraction compared with ECCO2R (112 ± 6 vs. 64 ± 5 ml/min, P < 0.001). Compared with baseline, R-ED and ECCO2R reduced minute ventilation by 50% and 27%, respectively. Systemic arterial gas analyses remained stable during the experimental phases. No major complication occurred, but there was an increase in creatinine level. CONCLUSIONS: In this first in vivo application, we proved electrodialysis feasible and effective in increasing membrane lung CO2 extraction. R-ED was more effective than conventional ECCO2R technology in controlling ventilation. Further studies are warranted to assess the safety profile of R-ED, especially regarding kidney function.

Conditioned medium from amniotic membrane-derived cells prevents lung fibrosis and preserves blood gas exchanges in bleomycin-injured mice-specificity of the effects and insights into possible mechanisms.

BACKGROUND AND AIMS: We recently demonstrated that injection of conditioned medium (CM) generated from cells of the mesenchymal region of human amniotic membrane (AMTCs) reduces bleomycin-induced lung fibrosis in mice, suggesting a crucial role of paracrine factor(s) secreted by AMTCs in these beneficial effects. We further investigated this hypothesis, the mechanisms involved, the effects on some lung functional parameters and whether AMTC-secreted effector(s) are specific to these cells and not produced by other cell types, extending the time of analysis up to 28 days after treatment. METHODS: Bleomycin-challenged mice were either treated with AMTC-CM or CM generated from human skin fibroblasts, human peripheral blood mononuclear cells or Jurkat cells, or were left untreated. Mouse lungs were analyzed for content of pro-inflammatory and pro-fibrotic molecules, presence of lymphocytes and macrophages and for fibrosis level (through histological semi-quantitative evaluation and quantitative measurement of collagen content). Arterial blood gas analysis was also performed. RESULTS: Up to 28 days after delivery, AMTC-CM-treated mice developed reduced lung fibrosis with respect to mice treated with other CM types. AMTC-CM-treated mice had comparatively better preservation of blood gas parameters and showed lower lung content of interleukin-6, tumor necrosis factor-α, macrophage inflammatory protein-1α, monocyte chemoattractant protein-1 and transforming growth factor-ß associated with reduced lung macrophage levels. CONCLUSIONS: AMTC-CM prevents lung fibrosis in bleomycin-challenged mice, improving survival and preserving lung functional parameters such as blood gas exchanges. The specificity of AMTC-CM action was indicated by the absence of fibrosis reduction when other CM types were used. Finally, we provide some insights into the possible mechanisms underlying AMTC-CM-mediated control of fibrosis.

Regional blood acidification enhances extracorporeal carbon dioxide removal: a 48-hour animal study.

BACKGROUND: Extracorporeal carbon dioxide removal has been proposed to achieve protective ventilation in patients at risk for ventilator-induced lung injury. In an acute study, the authors previously described an extracorporeal carbon dioxide removal technique enhanced by regional extracorporeal blood acidification. The current study evaluates efficacy and feasibility of such technology applied for 48 h. METHODS: Ten pigs were connected to a low-flow veno-venous extracorporeal circuit (blood flow rate, 0.25 l/min) including a membrane lung. Blood acidification was achieved in eight pigs by continuous infusion of 2.5 mEq/min of lactic acid at the membrane lung inlet. The acid infusion was interrupted for 1 h at the 24 and 48 h. Two control pigs did not receive acidification. At baseline and every 8 h thereafter, the authors measured blood lactate, gases, chemistry, and the amount of carbon dioxide removed by the membrane lung (VCO2ML). The authors also measured erythrocyte metabolites and selected cytokines. Histological and metalloproteinases analyses were performed on selected organs. RESULTS: Blood acidification consistently increased VCO2ML by 62 to 78%, from 79 ± 13 to 128 ± 22 ml/min at baseline, from 60 ± 8 to 101 ± 16 ml/min at 24 h, and from 54 ± 6 to 96 ± 16 ml/min at 48 h. During regional acidification, arterial pH decreased slightly (average reduction, 0.04), whereas arterial lactate remained lower than 4 mEq/l. No sign of organ and erythrocyte damage was recorded. CONCLUSION: Infusion of lactic acid at the membrane lung inlet consistently increased VCO2ML providing a safe removal of carbon dioxide from only 250 ml/min extracorporeal blood flow in amounts equivalent to 50% production of an adult man.

Inflammation and Behavior Changes in Dogs and Cats.

Sickness is a normal response to infections or stress triggered by proinflammatory cytokines that drive local and systemic inflammatory responses. Proinflammatory cytokines act on the brain causing the so called "sickness behavior,"which is thought to improve recovery but can become maladaptive in the long term. Chronic inflammation characterizes many diseases and there is some evidence that dogs and cats experience age-associated increases in inflammation, a condition named "inflammaging." A complex and multifactorial relationship exists between these inflammatory mechanisms, pain, and psychological illness that may complicate veterinary diagnosis and affect the outcome.

Infusion of 2.5 meq/min of Lactic Acid minimally increases CO2 production compared to an isocaloric glucose infusion in healthy anesthetized, mechanically ventilated pigs.

INTRODUCTION: Blood acidification by lactic acid infusion converts bicarbonate to CO2. This effect can be exploited to increase the transmembrane PCO2 gradient of an extracorporeal membrane lung, resulting in a significant increase of extracorporeal CO2 removal. Lactic acid, however, is an energetic substrate and its metabolism might increase total body CO2 production (VCO2), limiting the potential beneficial effects of this technique. The aim of our study was to compare VCO2 during isocaloric infusion of lactic acid or glucose. METHODS: Six pigs (45 ± 5 kg) were sedated and mechanically ventilated. Estimated caloric needs were 2,300-2,400 Kcal/die (95 to 100 Kcal/h). A sequence of two steps lasting four hours each was performed: 1) Glucose, 97 kcal/h were administered as 50% glucose solution, and 2) Lactic Acid, approximately 48.5 kcal/h were administered as lactic acid and approximately 48.5 kcal/h as 50% glucose solution. This sequence was repeated three times with two-hour intervals. Every hour VCO2, arterial blood gases and lactate were measured. Blood glucose level was kept constant by titrating an insulin infusion, ventilation was adjusted to maintain arterial PCO2 at 50 mmHg, a normal value for our animal model. RESULTS: During Lactic Acid steps VCO2 increased less than 5% compared to the Glucose steps (282 vs. 269 ml/min, P < 0.05); blood glucose did not differ between the two groups (respectively 101 ± 12 vs. 103 ± 8 mg/dl). Arterial lactate was always lower than 3 mmol/L. Arterial pH was lower during Lactic Acid steps (7.422 vs. 7.445, P < 0.05). CONCLUSIONS: Replacing 50% of the caloric input with lactic acid increased total CO2 production by less than 5% compared to an equal caloric load provided entirely by a 50% glucose solution.

The Relationship between Signs of Medical Conditions and Cognitive Decline in Senior Dogs.

Canine cognitive dysfunction syndrome (CCDS) is a progressive age-related neurodegenerative disorder in dogs. Minimal research has been performed to investigate how clinical signs may be impacted by other medical conditions. A cross-sectional study was performed using the Canine Cognitive Assessment Scale (CCAS) to evaluate cognitive impairment as reported by owners. Owner-reported health-related measures included behaviour changes, the body condition score, and veterinary diagnoses of disease. The responses from 804 dogs in the last 25% of their expected lifespan were analysed. Factors were identified in the owner-reported behavioural signs of disease representing pathologies in four body systems: musculoskeletal-neurological, digestive, metabolic, and dermatological, with the items comprising these factors also compiled into a cumulative measure of health. The results showed a strong correlation between the CCAS score and both the musculoskeletal-neurological factor and the overall cumulative measure of health. Moderate correlations between the CCAS score and the digestive factor and metabolic factor were also observed. The correlation between the dermatological factor and the CCAS score was weak. This study highlights the need to screen dogs for concurrent diseases when using scales to assess cognitive impairment and to monitor dogs who have health conditions, particularly those that are painful, for the onset of cognitive impairment.

Sniffer dogs performance is stable over time in detecting COVID-19 positive samples and agrees with the rapid antigen test in the field.

Rapid antigen diagnostic (RAD) tests have been developed for the identification of the SARS-CoV-2 infection. However, they require nasopharyngeal or nasal swab, which is invasive, uncomfortable, and aerosolising. The use of saliva test was also proposed but has not yet been validated. Trained dogs may efficiently smell the presence of SARS-CoV-2 in biological samples of infected people, but further validation is needed both in laboratory and in field. The present study aimed to (1) assess and validate the stability over a specific time period of COVID-19 detection in humans' armpit sweat by trained dogs thanks to a double-blind laboratory test-retest design, and (2) assess this ability when sniffing people directly. Dogs were not trained to discriminate against other infections. For all dogs (n. 3), the laboratory test on 360 samples yielded 93% sensitivity and 99% specificity, an 88% agreement with the Rt-PCR, and a moderate to strong test-retest correlation. When sniffing people directly (n. 97), dogs' (n. 5) overall sensitivity (89%) and specificity (95%) were significantly above chance level. An almost perfect agreement with RAD results was found (kappa 0.83, SE 0.05, p = 0.001). Therefore, sniffer dogs met appropriate criteria (e.g., repeatability) and WHO's target product profiles for COVID-19 diagnostics and produced very promising results in laboratory and field settings, respectively. These findings support the idea that biodetection dogs could help reduce the spread of the virus in high-risk environments, including airports, schools, and public transport.

Physiotherapy Improves Dogs' Quality of Life Measured with the Milan Pet Quality of Life Scale: Is Pain Involved?

Quality of life is defined as an individual's satisfaction with its physical and psychological health, its physical and social environment, and its ability to interact with the environment. Understanding companion dogs' QOL can help veterinarians and owners know when treatment options have successfully alleviated symptoms of disease in such fields as veterinary physiotherapy. For this study, 20 adult dogs were selected from patients of a physiotherapy referral center with orthopedic, neurological, and/or degenerative conditions. The severity of the medical problem was ranked, and the symptoms, the treatment plan, and demographic data were recorded at the time of the physical examination. In addition, the owner of the dog was asked to fill out a questionnaire on the quality of life of the pet (the Milan Pet Quality of Life scale) at the time of the first consultation as well as the last follow-up after the treatment. The MPQL measures four domains of QOL: physical (signs of medical conditions), psychological (emotional and behavioral well-being), social (quality and extent of social interactions), and environmental (freedom and safety in one's environment). The results of the study indicated a significant improvement in the psychological QOL domain following physiotherapeutic treatment. The social QOL domain declined with the severity of lameness, while the physical QOL, as reported by the owner, declined with the overall criticality of the medical condition, as ranked by the physiotherapist. The results of the study support the recent evidence of a relationship between pain and canine psychological well-being and highlight the importance of investigating psychological and emotional aspects of dogs' QOL when treating orthopedic and neurological cases with physiotherapy.

Signs of Anxiety and Salivary Copeptin Levels in Dogs Diagnosed with Separation-Related Problems in a Short Separation Test.

The need for faster diagnosis and more accurate treatment decisions in separation-related problems (SRPs) in dogs is urgent, and a more precise behavioral phenotyping and the development of biomarkers may be of great value. Vasopressin could be a potential non-invasive biomarker of anxiety in dogs with SRPs, but reliable measurement of its concentration is challenging. Here, we compared the behavior and salivary concentrations of copeptin, an arginine vasopressin surrogate, in dogs with SRPs (Case group, n = 13) and with no problems (Control group, n = 15) as they were introduced to a novel environment and subjected to a short episode of separation and reunion with the owner. Dogs in the Case group had greater odds of showing locomotory or oral behaviors during the pre- and post-separation than Controls, while the odds were significantly lower during separation. They also had greater odds of being persistent in seeking attention and proximity from the stranger during reunion. Overall, dogs with SRPs were more likely to express an anxiety-like state during the entire test than Controls, with separation from the owner, and even its anticipation, possibly accounting for this group difference. Although salivary copeptin concentrations did not differ between the two groups, a different trend was detected in Cases and Controls that is worth exploring in further validation studies involving a larger sample.

Changes in the lipidome of water buffalo milk during intramammary infection by non-aureus Staphylococci.

This study aimed to determine the lipidome of water buffalo milk with intramammary infection (IMI) by non-aureus staphylococci (NAS), also defined as coagulase-negative staphylococci, using an untargeted lipidomic approach. Non-aureus Staphylococci are the most frequently isolated pathogens from dairy water buffalo milk during mastitis. A total of 17 milk samples from quarters affected by NAS-IMI were collected, and the lipidome was determined by liquid chromatography-quadrupole time-of-flight mass spectrometry. The results were compared with the lipidome determined on samples collected from 16 healthy quarters. The study identified 1934 different lipids, which were classified into 15 classes. The abundance of 72 lipids changed in NAS-IMI milk compared to healthy quarters. Significant changes occurred primarily in the class of free fatty acids. The results of this study provided first-time insight into the lipidome of dairy water buffalo milk. Moreover, the present findings provide evidence that NAS-IMI induces changes in water buffalo milk's lipidome.

Peptidomic changes in the milk of water buffaloes (Bubalus bubalis) with intramammary infection by non-aureus staphylococci.

Mastitis by non-aureus staphylococci (NAS) is a significant issue in dairy buffalo farming. In a herd with subclinical NAS mastitis, we identified Staphylococcus microti as the predominant species. To assess milk protein integrity and investigate potential disease markers, we characterized 12 NAS-positive and 12 healthy quarter milk samples by shotgun peptidomics combining peptide enrichment and high-performance liquid chromatography/tandem mass spectrometry (LC-MS/MS). We observed significant changes in the milk peptidome. Out of 789 total peptides identified in each group, 49 and 44 were unique or increased in NAS-positive and healthy milk, respectively. In NAS-positive milk, the differential peptides belonged mainly to caseins, followed by milk fat globule membrane proteins (MFGMP) and by the immune defense/antimicrobial proteins osteopontin, lactoperoxidase, and serum amyloid A. In healthy milk, these belonged mainly to MFGMP, followed by caseins. In terms of abundance, peptides from MFGMP and immune defense protein were higher in NAS-positive milk, while peptides from caseins were higher in healthy milk. These findings highlight the impact of NAS on buffalo milk quality and mammary gland health, even when clinical signs are not evident, and underscore the need for clarifying the epidemiology and relevance of the different NAS species in this dairy ruminant.

Domestic dogs (Canis familiaris) grieve over the loss of a conspecific.

Behavioural reactions towards a dead conspecific have been observed rarely in wild canids and there is no documented scientific evidence of grief in pet dogs. A quantitative analysis of grief-related responses in both dogs and owners was conducted, using the validated online Mourning Dog Questionnaire. The survey was completed by 426 Italian adults who had owned at least two dogs, one of whom died while the other was still alive. This research aims to explore whether, how and what a dog may experience over the loss of a companion dog. Multiple logistic regression indicates that both a friendly or parental relationship between two dogs but also the fact that dogs used to share food and the owner's grief and anger are principal predictors of negative behavioural changes. According to dog owners' answers, the surviving dog after the death of the companion dog changed both in terms of activities ("playing", "sleeping", and "eating") and emotions (fearfulness), which occurred as a function of the quality of the relationship between the two animals. By contrast, the time the two dogs had spent together had no effect on the behaviours of surviving dog. Owner perceptions about their dog's reactions and emotions were not related to the memory or suffering of the event that tended to diminish over time. These findings indicate that a dog may show grief-related behavioural and emotional patterns when a close conspecific dies, with aspects of the latter possibly related to the owner's emotional status.

The Cat-Owner Relationship: Validation of the Italian C/DORS for Cat Owners and Correlation with the LAPS.

Globally, most people now own a pet. Scales have been developed to understand the impact of pet ownership on people's lives and to measure specific aspects of the owner-pet relationship. The Cat-/Dog-Owner Relationship Scale (C/DORS) is a tool developed to investigate this relationship in both dog and cat owners. The aim of the study was to refine and validate the C/DORS for cat owners in Italian. Exploratory Factor Analysis and Confirmatory Factor Analysis (CFA) were used to determine the most appropriate factor model. Construct validity was confirmed by correlating the C/DORS subscales with the Lexington Attachment to Pets (LAPS) subscales. Results confirmed the original structure of the English version (i.e., three factors: Pet Owner Interaction = POI, Perceived Emotional Closeness = PEC, Perceived Costs = PC) and CFA confirmed the structure of LAPS and C/DORS scales. Cronbach's alpha demonstrated the Italian version of the two scales to have good internal reliability for all domains. Owners of cats living exclusively indoors reported higher scores on POI and PEC compared to indoor/outdoor cats. Owning both cats and dogs was correlated with lower scores on POI, and fewer perceived costs (i.e., PC) of cat ownership. Finally, behaviour problems, not being neutered, and lack of previous experience with cat ownership were associated with lower scores on PC.

Mechanical ventilation and volutrauma: study in vivo of a healthy pig model.

Mechanical ventilation is essential in intensive care units. However, it may itself induce lung injury. Current studies are based on rodents, using exceptionally large tidal volumes for very short periods, often after a "priming" pulmonary insult. Our study deepens a clinically relevant large animal model, closely resembling human physiology and the ventilator setting used in clinic settings. Our aim was to evaluate the pathophysiological mechanisms involved in alveolo/capillary barrier damage due to mechanical stress in healthy subjects. We randomly divided 18 pigs (sedated with medetomidine/tiletamine-zolazepam and anesthetised with thiopental sodium) into three groups (n=6): two were mechanically ventilated (tidal volume of 8 or 20 ml/kg), the third breathed spontaneously for 4 hours, then animals were sacrificed (thiopental overdose). We analyzed every 30' hemogasanalysis and the main circulatory and respiratory parameters. Matrix gelatinase expression was evaluated on bronchoalveolar lavage fluid after surgery and before euthanasia. On autoptic samples we performed zymographic analysis of lung, kidney and liver tissues and histological examination of lung. Results evidenced that high Vt evoked profound alterations of lung mechanics and structure, although low Vt strategy was not devoid of side effects, too. Unexpectedly, also animals that were spontaneously breathing showed a worsening of the respiratory functions.

Use of the Milan Pet Quality of Life Instrument (MPQL) to Measure Pets' Quality of Life during COVID-19.

The COVID-19 pandemic caused lifestyle changes, with unknown effect on pets' quality of life (QoL). Between May and July 2020, we distributed an online survey to investigate the role of several factors on feline and canine QoL, including lockdown-related factors. We used existing scales to measure human and pets' personalities (Reinforcement Sensitivity Theory Personality Questionnaire, RST-PQ; RST-Dog; RST-Cat) and the human-animal relationship (Lexington Attachment to Pets Scale, LAPS) and the Milan Pet Quality of Life instrument (MPQL). Overall, 235 participants reported about 242 adult pets (Ncats = 78, Ndogs = 164). Factor analysis confirmed the structure and internal reliability of the existing scales (RST-PQ, RST-Dog, RST-Cat, LAPS) and suggested a four-factor structure for the MPQL (physical, psychological, social, environmental). The results indicate that the pets' psysical QoL was largely explained by pet-related elements (pets' demographics and life experience, and pets' personality). Conversely, the pets' psychological QoL was explained mostly by owner-related elements, such as the owners' demographics, COVID-19-related changes, and the owners' personality. Predictably, the pets' environmental QoL is mostly explained by environmental factors, such as the outdoor access in the home environment and the country. Finally, the pets' social QoL was explained by the larger combination of models: pets' characteristics and personality, environment and COVID-19-related changes, and the pet-human relationship. These findings can be explained by two non-mutually exclusive mechanisms. The reported changes may be a by-product of the COVID-19 pandemic's psychological and lifestyle effects on the owners, which in turn alter the way the owners interact with their pets and look after them. However, the owners' characteristics and mood may bias their answers regarding their pets.

Personality and Cognitive Profiles of Animal-Assisted Intervention Dogs and Pet Dogs in an Unsolvable Task.

Dogs are biologically predisposed to communicate with humans in cooperative contexts. They show individual differences in dog-human communication and inhibition, potentially enhanced by life experience, e.g., Animal-Assisted Interventions. This study aimed to investigate whether dogs' personality, defined by biologically meaningful neural circuits described in the Reinforcement Sensitivity Theory of Personality (RST), predicted dogs' communication, task orientation, emotional state, and approach of an unsolvable task. We also investigated the differences between dogs experienced in Animal-Assisted Interventions (AAI) and inexperienced dogs. The results indicated that a high sensitivity to the RST personality trait related to managing uncertainty (Behavioural Inhibition System, BIS) predicted fewer task orientations but increased referential and non-referential looking, which we interpreted as a way to obtain directions from the owner. Conversely, a high sensitivity to the traits reflecting tendencies to approach rewards (Behavioural Approach System, BAS) and avoid punishment (Fight-Flight-Freeze System, FFFS) predicted lower looking. High sensitivity to the FFFS also predicted more frequent task orientations, which we interpreted as frustration. Finally, the dogs in the AAI program looked more at their owner and were less oriented towards the task. These results provide empirical evidence of individual differences tied to the psychobiological personality traits in canine cognitive skills. Understanding such cognitive profiles may have an enormous impact on activities that rely upon dog-human collaborative interaction.

Influence of subclinical mastitis and intramammary infection by coagulase-negative staphylococci on the cow milk peptidome.

Coagulase-negative staphylococci (CNS) are the most prevalent microorganisms isolated from cow milk and are associated with subclinical mastitis and persistent increases in the bulk milk somatic cell count (BMSCC) of low BMSCC herds. By combining peptide enrichment, LC-ESI-MS/MS, and statistical analysis, we investigated the influence of subclinical mastitis and CNS infection on the milk peptidome. Quarter milk samples from clinically healthy Holstein cows were subjected to bacteriological culture (BC) and somatic cell counting (SCC) for two consecutive samplings and 28 (including 11 negatives and 17 positives) were selected for peptidomic analysis. The study identified 1363 different endogenous peptides and highlighted a significant increase of peptides in CNS-positive milk, mainly represented by casein fragments. Milk peptidome changes increased with the SCC, as also demonstrated by protein electrophoresis and densitometry. Peptides significantly different in CNS or CONTROL samples were identified and characterized. Our results indicate that subclinical mastitis by CNS can induce significant changes in the milk peptidome, opening the way to future studies for the identification of a biomarker panel as well as for the understanding of their consequences for the technological and sensorial characteristics of cow milk and dairy products. SIGNIFICANCE: This is the first investigation on the impact of subclinical CNS mastitis on the bovine milk peptidome. The peptide enrichment strategy combined with a highly sensitive MS/MS analysis enabled the compilation of a very large peptide dataset for healthy and mastitic milk. The comparison of CNS and Control samples, also considering SCC classes, highlighted several peptides with potential for understanding milk protein and peptide dynamics in subclinical mastitis, with possible implications for its detection.

Salivary Vasopressin as A Potential Non-Invasive Biomarker of Anxiety in Dogs Diagnosed with Separation-Related Problems.

Physiological biomarkers of canine anxiety have not been extensively investigated to date. To identify new biomarkers in dogs, we compared behaviorally normal dogs (Control group, N = 13) to dogs diagnosed with separation problems (Case group, N = 13) as they were introduced into a novel environment in the presence of two strangers and subjected to a short episode of separation and reunion with the owner. During the separation phase, dogs in the Case group explored significantly less than controls and were significantly more persistent in expressing passive stress-coping strategies aimed at seeking proximity to their owners. When the owners returned, dogs with separation distress spent significantly more time jumping up on the strangers than control dogs did. Salivary oxytocin and vasopressin concentrations did not differ between samples taken before and after the separation. However, vasopressin concentrations immediately after separation were significantly higher in the Case than in the Control group and remained higher, although not significantly so, 10 minutes later. These results indicated that dogs with separation distress became more anxious than typical dogs when separated from their owner in an unfamiliar environment and provided preliminary support for the use of salivary vasopressin as a possible biomarker for anxiety-related responses in dogs.

The Impacts of a Reading-to-Dog Programme on Attending and Reading of Nine Children with Autism Spectrum Disorders.

Poor knowledge is available on the effectiveness of reading to dogs in educational settings, particularly in children with Autism Spectrum Disorders (ASD). In this study, we test the hypothesis that reading to a dog improves propensity towards books and motivation to read after the end of the programme, as well as reading and cognitive skills in children with ASD. The study is a prospective, randomized controlled trial, consisting of testing and re-testing after a 10 sessions reading programme with and without the presence of a dog. Nine Children with ASD (6-11 years old) were randomly assigned to a control (CG, reading without a dog, n. 4) or experimental group (EG, reading to a dog, n. 5). Children's attendance at reading sessions was recorded at each session. Parents' perceptions were evaluated at the end of the programme to detect changes in children's attitudes and motivation toward reading. Psychologist-administered validated reading (Cornoldi's MT2 reading test; test of reading comprehension, TORC; metaphonological competence test, MCF) and cognitive tests (Wechsler intelligence scale for children Wisc IV, Vineland) to all children, at baseline and at the end of the reading programme. Compared with CG children, children in the EG group participated more frequently in the reading sessions, and they were reported to be more motivated readers at home after the programme. However, there were no differences on reading and cognitive tests' scores either within each group of children or between groups. Further studies are warranted in order to understand whether and how incorporating dogs into a reading programme is beneficial to Children with ASD at the socio-emotional and cognitive level.