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The aim of this study was to assess the changes of pro-inflammatory interleukins in 10 horses subjected to road transport practices (distance of 150 km) from the training site (Messina, Sicily) to the competition centre in Syracuse (Sicily). Blood sampling and interleukins analysis were performed during a round trip transportation (transport 1 and transport 2). In particular, blood samples were collected before the transport took place (Pre), five minutes later (Post) and one hour later (Post 1 h), for each transport, in order to assess the serum concentration of IL-1α, IL-1β, IL-2 and IL-6. The results showed that the serum concentration of IL-1α decreased at Post and Post 1 h compared to the values obtained at rest condition (P < 0.05). The other interleukins analysed (i.e. IL-1β, IL-2 and IL-6) showed increased levels at Post than Rest and Post 1 h in transport 1 (P < 0.05). In transport 2 the analysed parameters showed no change throughout the analysed time points (P > 0.05); however, higher levels of IL-1α at Pre and higher IL-1β, IL-2 and IL-6 values at Post were found in transport 1 than transport 2 (P < 0.05). The increase in pro-inflammatory cytokines after transport 1 suggests the triggering of the inflammatory event and this may show that, although horses are animals accustomed to transport, this is a stressful event that could activate the well-orchestrated inflammation cascade, albeit physiological and temporary, as highlighted by the lower serum concentrations of the investigated interleukins found in transport 1 than transport 2 and by the lack of significant differences in the serum concentrations of the investigated interleukins among the time points of transport 2. It must be taken into account that enrolled animals are well-trained and healthy athletic horses participating to a jumper competition, thus, such inflammation did not occur thanks to a good balance between pro-inflammatory and anti-inflammatory cytokines which allowed a prompt restoration of homeostasis eventually impaired by the stressful event., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Due to the paucity of information on circus management effects on the welfare of horses, this study investigated the plasma concentrations of noradrenaline, dopamine and serotonin, known to be indices of mental status, as well as the reactive oxygen metabolites (d-Roms) and the biological antioxidant potential (BAP), likely to denote the oxidant/antioxidant equilibrium of organisms, in horses managed in different Italian circuses. For the study, 56 circus horses of different breeds and ages were enrolled and divided into six groups according to the horses' management (circus management, groups G1-G5; classic riding management representing the control group, CG). From each horse, blood samples were collected in order to assess the concentration of selected parameters. One-way ANOVA showed no differences ( p > 0.05) in serotonin, dopamine, noradrenaline, d-Roms and BAP values between circus and control horses. No differences related to the breed of the horses enrolled in the study were found in the values of all investigated parameters ( p > 0.05). Furthermore, neurotransmitters showed overlapping levels between the different age classes of investigated horses ( p > 0.05); contrariwise, the age of the horse displayed a significant effect on BAP values, with the oldest horses (16-21 age class) exhibiting lower BAP values compared to 4-5, 6-10 and 11-15 age classes ( p < 0.05), whereas the d-Roms showed similar values in horses of different age classes ( p > 0.05). The results gathered in the present study suggest that the mental status of horses under circus management was not compromised; however, better attention and care in the management of older horses is advocated, as they showed a lower biological antioxidant potential than younger horses; thus, they could be more susceptible to oxidative stress.

This study aimed to evaluate changes in body temperature in athletic horses during two different road transport distances. Six Italian Saddle horses were subjected to a 100 and 300 km transport during different times of day (am and pm). Rectal and cutaneous temperatures were recorded before (T0), immediately (following 5 min- T1) and 1 hour (T2) after transport by means of a rectal digital thermometer and a thermal infrared camera (FLIR T440) respectively, for the evaluation of left and right side of four body regions: jugular, shoulder, croup and inner thigh. There were no differences between left and right sides, inner thigh or rectal temperatures when comparing the transport distance, time points or time of day. At T0, jugular (P < 0.0001), shoulder (P < 0.01) and croup (P < 0.01) average temperatures were higher in the pm compared to those in the am in both journeys. At T1, jugular (P < 0.01) and croup (P < 0.01) temperatures were lower in the pm compared to am following the 300 km journey. Jugular temperature (P < 0.0001) was higher following the 300 km compared to the 100 km journey at each time point (T1 and T2) at both times of day (am and pm). Shoulder (P < 0.0001) and croup temperatures (P < 0.0001) were higher at T2 after the 300 km journey than at T2 after the 100 km journey). The current results suggested a difference between the two distances and the time of day appeared to have as great effect on ocular temperature as road transport distance., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The aim of the present study was to investigate eye temperature modifications after road transport in athletic horses habituated to travel. Eight adult Italian saddle horses traveled 100 km and, two weeks later, 300 km. Eye temperature (ET), rectal temperature (RT) and serum cortisol concentration were assessed before (T1), after (T2) and 60 min (T3) after the road transport. ET was evaluated with infrared thermography (IRT) in three regions of interest: EL1 (medial canthus), EL2 (central cornea) and EL3 (lateral canthus). Two-way analysis of variance (ANOVA) for repeated measures showed statistically higher values at T2 and T3 for EL1 ( p < 0.01), EL2 ( p < 0.01) and EL3 ( p < 0.01) following the 100 km journey. RT ( p < 0.01) showed higher values at T2 and T3 after the 100 km journey and higher values at T2 ( p < 0.01) following the 300 km journey. ET values were positively correlated with RT at T1, T2 and T3 following the 100 km journey and at T2 following the 300 km journey and positively correlated with serum cortisol concentration at T1, T2 and T3 following the 100 km journey and at T2 and T3 following the 300 km journey. Eye temperature monitoring with IRT allows quick and practical strategies to monitor an animal's physiological state and welfare during daily activities.

This review highlights recent findings on biological rhythms and discusses their implications for the management and production of domestic animals. Biological rhythms provide temporal coordination between organs and tissues in order to anticipate environmental changes, orchestrating biochemical, physiological and behavioural processes as the right process may occur at the right time. This allows animals to adapt their internal physiological functions, such as sleep-wake cycles, body temperature, hormone secretion, food intake and regulation of physical performance to environmental stimuli that constantly change. The study and evaluation of biological rhythms of various physiological parameters allows the assessment of the welfare status of animals. Alteration of biological rhythms represents an imbalance of the state of homeostasis that can be found in different management conditions.

Introduction: The objective of the research was to investigate the effect of Saccharomyces cerevisiae supplementation on some acute-phase proteins, haptoglobin and all electrophoretic parameters in young Charolaise bulls., Material and Methods: Sixty bulls were divided into two equal groups: the control group (CG) receiving the base diet without yeast supplementation and the diet supplementation group (YG) receiving the base diet with 5g of Saccharomyces cerevisiae supplementation. The base diet was total mixed ration allocated at 11.85 kg per animal per day. Blood samples were collected from all bulls on day 0 before the start of the diet supplementation, and on days 20 and 40 after the start. Total proteins, albumin, globulin fraction (α1-, α2-, β1-, β2- and γ-globulins), albumin: globulin ratio (A: G) and haptoglobin were determined., Results: Two-way analysis of variance showed a significant effect of the yeast feeding time on all studied parameters except α2-globulins in both groups. The YG showed a higher average concentration of total proteins, albumin and A: G and a lower average concentration of γ-globulins and haptoglobin than the CG., Conclusion: These results indicated the beneficial effect of the Saccharomyces cerevisiae on the inflammatory status of the young bulls, which showed an adequate response in serum levels of the acute-phase proteins tested., Competing Interests: Conflict of Interests Statement: The authors declare that there is no conflict of interests regarding the publication of this article., (© 2024 Francesca Arfuso et al., published by Sciendo.)

Background: This research aimed to evaluate the association between the monthly consumption of fish (differentiated by type) and both gestational and neonatal outcomes. Methods: Women who were admitted for delivery in the last 6 months of 2023 were prospectively included and divided according to type of fish consumed (based on DHA and mercury content) and frequency of consumption. Neonatal outcomes included weight, length, head circumference, and 1st and 5th minute Apgar scores. Maternal outcomes were threats of abortion, preterm birth, gestational diabetes and hypertension, cesarean section, and differential body mass index (BMI). Results: Small-size oily fish with high DHA and low mercury content (type B fish) consumption was positively associated with neonatal weight and head circumference, and less weight gain in pregnancy. It was also significantly associated with lower incidences of gestational diabetes and hypertension, and cesarean section. Correlation between differential BMI and monthly consumption of fish resulted in statistical significance, especially in type B fish consumers. Conclusions: The consumption of type B fish was significantly associated with increased neonatal weight and head circumference and better maternal outcomes.

Background and Purpose: Biological monitoring of trace element horses is a well-known tool for investigating potential bioaccumulation in urbanized and industrialized geographical areas. Some biomaterials such as hoof are considered as an important indicator of environmental pollution. Hooves can store trace elements for a long time compared to blood and this represents a scientific key to long-term monitoring of exposure to environmental pollutants. In the present study, samples of equine hoof and blood were taken from an experimental group of horses living in an industrialized area of Sicily (Italy) and from a control group of horses housed in a non-industrialized area of Sicily to evaluate the bioaccumulation of different trace elements in different substrates., Methods: Hoof and blood mineral concentration of Li- Lithium, Cu- Copper, Zn- Zinc, Sr- Strontium and, Pb- Lead was processed by means of Thermo Scientific iCAP-Q ICP-MS spectrometer. Student T-test was applied to evaluate the differences between groups and substrates. Correlation analysis between substrates and hematological parameters was performed., Results: Statistical analysis showed a significant statistical difference for Li (p < 0.0001) and Pb (p < 0.0001) between blood and hoof in both groups. A statistically significant difference was observed for Li (p < 0.0001), Cu(p < 0.01), Zn (p < 0.0001) and Pb (p < 0.0001) between substrates in the experimental group. The statistical analysis showed a significant difference of blood Zn (p < 0.01) and hoof Pb (p < 0.01) concentration between groups. Our results revealed a statistically higher concentration of minerals in the blood substrate than in the hoof in both groups. Li (p < 0.0001) and Zn (p < 0.001) were positively correlated between control and experimental group in blood substrate. No correlations were found (P > .05) in hoof between groups for other elements. A positive correlation with WBC was observed in Cu concentration for hoof substrate (p < 0.02) in control group and for Zn in experimental group (p < 0.02). A positive correlation with HGB and HCT was observed for Cu hoof concentration (p < 0.02) in experimental group and PLT resulted positively correlated (p < 0.02) with Pb blood substrate in control group., Conclusions: Horses were not overexposed to these elements in the analyzed groups. The study highlights the role of a new bioaccumulation substrate such as the horse hoof identified as an analytical matrix for the monitoring of heavy metal concentrations in domestic animals., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Dogs are the common pets adopted by humans, and their circadian behavior and physiology are influenced by human habits. In many families, there is a change of lifestyle with respect to the natural daylight (NDL) cycle. Exposure to constant light disrupts some central and peripheral circadian rhythms. The aim of the present study was to improve the knowledge about the circadian changes of clock components in the peripheral blood in dogs housed under NDL and constant light (LL) conditions. Blood samples were collected on five female Beagle dogs (2 years old, 14 ± 0.5 kg) every 4 hours for a 24-hour period during an NDL (Sunrise 05:05 h - Sunset 20:55  h) and 24-hour period of constant light (LL). Blood samples were stored in a PAX gene Blood RNA Tube, real-time RT-quantitative polymerase chain reaction was performed to determine Clock, Per1-3, and Cry1-2 gene expression. During the NDL, all genes investigated showed robust diurnal daily rhythmicity. During the constant light, only Clock maintained its daily rhythmicity. Clock acrophase was observed close to sunrise (ZT 0) and was statistically different from the other clock genes except for Per3 . Per3 daily oscillations were not statistically significant. No differences were observed among the clock genes tested in the amplitude and robustness values. Our results can be considered preliminary data to provide new insights into the adaptation mechanism of the canine peripheral circadian clock. The persistence of Clock gene expression during the LL indicated the presence of an endogenously generated signal in blood. Because peripheral blood is an easily accessible sample in dogs, the analysis of clock gene expression in this tissue could be useful to investigate the adaptive capacity of this species housed in different environmental conditions linked to the owner's lifestyle.

This study aimed to investigate the common social and communicative behaviors of the Fonni's Dog under different outdoor conditions. For this study, 70 adult dogs (3-7 years; 32 intact males, 38 intact females) belonging to the Fonni's breed were used. A total of 35 dogs were kept in kennels and 35 were free-ranging dogs in their sheep/goat livestock units. A behavioral repertoire was adapted from the literature and an ethogram was filled in for each dog. All dogs were evaluated in the presence of the owner. Fisher's exact test, following Bonferroni's correction, was used to test possible differences in the categorical variables (presence or absence of the behavior) between free-ranging dogs and dogs kept in kennels. The study revealed that several categories of the dogs' body language were associated with the management condition. However, the breed motivations (guarding and defense of the territory) were satisfied both in kennel and in the animals who were free in the property. The current study suggests a good behavioral balance of the Fonni's Dogs which could be attributed to correct communication between dogs and owners.

In Italy, the consumption of pork meat is increasing, despite consumers' attitudes being addressed toward a greater sensitivity about animal welfare and its link with safe food. Considering the relatively high animal welfare standards and the divergence in public interest in farm animal welfare and ethical issues, the objective of this study-in continuation of our previous paper relating to consumer behavior and preferences in welfare-friendly pork breeding-was to investigate habits of pork consumers regarding pig welfare, principally evaluating their willingness to pay (WTP) a higher price for pork obtained using raising techniques with an approach based on animal welfare. An ad hoc questionnaire-based survey was submitted to consumers (n = 404) in Messina province, Italy. Results suggest that 47% of consumers were willing to pay an additional price for pork from farms that apply specific animal welfare standards. Positive correlations were between WTP and farming techniques ( p = 0.001), organic farming methods ( p = 0.001), and farms in which animal welfare is taken care of and guaranteed ( p < 0.001). These findings suggest that consumers intend to pay a higher price for pork, like other animal products obtained using animal-friendly raising techniques. The sensitivity to the animal welfare of a single human being may influence consumers' attitudes toward pork consumption.

Cardiac biomarkers are useful to identify cardiac muscle variations in human and equine medicine. The aim of this study was to investigate the acute effect of a bout of show jumping training on serum activity of cardiac and muscular biomarkers in healthy athletic horses to include cardiac troponin (cTnI), myoglobin (MB), aspartate amino transferase (AST), alanine amino transferase (ALT), creatine phosphokinase (CPK) and lactate dehydrogenase (LDH). Serum samples were collected from seven Italian Saddle horses (three geldings and four mares; 10 ± 3 years; mean body weight 480 ±70 kg), regularly trained for show jumping at rest, immediately after exercise (show jumping simulate trial) and during the recovery period (30 and 60 min after exercise). ANOVA was applied to all parameters, and Pearson correlation coefficient t (r) evaluated. Immediately after exercise there was an increase in cTnI (P < .01), MB (P < .01), and CPK (P < .005); a positive correlation between cTnI and AST and between AST and LDH; and a negative correlation between cTnI and ALT and between ALT and CPK. Thirty minutes after exercise, there was a positive correlation between AST and ALT and between AST and LDH, while 60 mintues after exercise, there was a positive correlation between MB and LDH and a negative correlation between AST and CPK. The results obtained demonstrate the cardiac and muscular response to short-term intense exercise show jumping exercise., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors., (Copyright © 2023 Elsevier Inc. All rights reserved.)

This study investigates consumer behavior and interest in "welfare-friendly" forms of pork production, considering the growing presence of animal welfare-focused breeding practices. The aim is to outline the typical profile of pork consumers and identify the key attributes influencing their purchasing decisions. A survey was conducted on a sample of 286 individuals after excluding those who only consumed beef and/or poultry or identified as vegetarians/vegans. Regression coefficients (b), 95% Confidence Intervals (95% C.I.), and p -values were reported for univariate and multivariate models. Statistical significance was determined at p < 0.05 (indicated in bold). The findings indicate that younger participants show greater sensitivity towards consuming meat raised using welfare-friendly methods, raising considerations about the age composition of the sample. The research's originality lies in evaluating consumer interest in pork raised with animal welfare-respecting techniques. The use of appropriate statistical tools, such as multivariate and multilayer models, allows effective solutions for multidimensional hypothesis testing problems in non-parametric permutation inference.

The evaluation of the performance levels in athletic horses is of major importance to prevent sports injuries. Raman spectroscopy is an innovative technique that allows for a rapid evaluation of biomolecules in biological fluids. It also permits qualitative and quantitative sample analyses, which lead to the simultaneous determination of the components of the examined biological fluids. On the basis of this, the Raman spectroscopy technique was applied on serum samples collected from five Italian Saddle horses subjected to a standardized obstacle course preceded by a warm-up to evaluate the applicability of this technique for the assessment of central and peripheral fatigue in athletic horses. Blood samples were collected via jugular venipuncture in a vacutainer tube with a clot activator before exercise, immediately after exercise, and 30 min and 1 h after the end of the obstacle course. Observing the obtained Raman spectra, the major changes due to the experimental conditions appeared in the (1300-1360) cm -1 and (1385-1520) cm -1 bands. In the (1300-1360) cm -1 band, lipids and tryptophan were identified; in the (1385-1520) cm -1 band, leucine, glycine, isoleucine, lactic acid, tripeptide, adenosine, and beta carotene were identified. A significant effect of exercise was recorded on all the sub-bands. In particular, a change immediately after exercise versus before exercise was found. Moreover, the mean lactic concentration was positively correlated with the Raman area of the sub-band assigned to lactic acid. In this context, the application of Raman spectroscopy on blood serum samples represents a useful technique for secondary-structure protein identification to investigate the metabolic changes that occur in athletic horses during physical exercise.

Inflammaging or chronic, low-grade inflammation is a phenomenon characterizing age-related pathologies and natural processes in aging tissues. This study aimed to investigate the effect of age on the serum levels of C-reactive protein and protein electrophoretic pattern in horses. Thirty Italian Saddle horses from the same horse training center, were enrolled and divided in three equal groups according to their age: Group A (2-4 years), Group B (7-10 years), Group C (15-20 years). Blood samples were collected from each horse to evaluate hematological parameters and the serum concentration of C-reactive protein (CRP), total proteins, albumin, α1-, α-2, β1-, β2- and γ-globulins. One-way analysis of variance showed an age-related difference in the concentration of CRP, α1- and α2-globulins (P < 0.001) which increased with aging. Albumin and albumin:globulin ratio showed lower values in Groups B and C than Group A (P < 0.001). Age of horses showed a negative correlation with the values of albumin, and a positive correlation with CRP, α1- and α2-globulins concentration. The results of the current study suggest an increased acute phase response in adult and old horses compared to young ones. As α-globulin fractions include many other acute phase proteins (APPs) in addition to CRP, further studies to assess what other APPs could be involved in chronic inflammation or "inflammaging" are recommended in horse in order to improve knowledge on the key inflammatory biomarkers during aging in this species., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier Inc. All rights reserved.)

During husbandry, domestic animals are exposed to many factors that can influence their circadian physiology organization leading to an increase in animals' discomfort. Thermal homeostasis is at the basis of animal wellness, the aim of the present study was to investigate the daily fluctuation of serum concentrations of 3,5,3'-triiodothyronine (T3) and thyroxine (T4) in association with the daily fluctuation of uncoupling protein 1 (UCP1) and clock gene Per2 in healthy horses housed in individual box, to improve the knowledge on this matter. Seven clinically healthy female Italian Saddle horses (8-10 years old, 510 ± 32 kg), were housed in individual boxes under natural photoperiod and environmental temperature and humidity. Blood samples were collected at 4-hour intervals over a 48-hour period, for the assessment of T3, T4, UCP1, and clock gene Per2. The application of two-way analysis of variance (ANOVA) on raw data showed a statistically significant effect of time of day on all studied parameters. A robust daily rhythm of T3, T4, and Per2 was observed. T3 showed a diurnal rhythm, with the acrophase at about 5 hours after sunrise, T4 acrophase was observed in the middle of the scotophase, Per2 acrophase was observed close to sunrise. In conclusion, we can claim that in horses kept under natural environmental conditions and not subjected to thermal stress, there is a daily rhythm of thyroid hormones associated with a daily rhythm of Per2 expression in the peripheral blood, and UCP1 remained constant during the two days., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

This study aimed to evaluate the change of white blood cell count, serum concentration of cortisol, C-reactive protein, albumin and globulin fractions in horse after road transport, and to assess the linkage among hypothalamic-pituitary-adrenal axis (HPA) and inflammatory reaction. From 10 horses blood samples were collected at rest, before road transport (218 km) (BT); after unloaded (AT), 30 and 60 min after unloaded (AT30 and AT60) in order to assess white blood cell count (WBC), serum cortisol, C-reactive protein (CRP), total proteins, albumin, α1-, α-2, β1-, β2- and γ-globulins. WBC, cortisol, CRP, α1-, α-2 and β2-globulins values increased after road transport than rest condition (p < 0.001). Albumin and A/G ratio showed lower values after road transport than rest (p < 0.001). Pearson's test showed a negative correlation between cortisol and the values of WBC, CRP, α1-, α2-, β1-, β2- globulins, and a positive correlation between WBC and serum concentration of CRP, α1- and α2-, β1-, β2-globulins at AT and AT30. The results showed that road transport evokes an inflammatory like-status in horses. Moreover, the activation of HPA and the onset of acute phase reaction in response to road transport seem to be interconnected with effects on horse's immune status., (© 2023. The Author(s).)

The aim of our study was to analyze circadian rhythm of the hematological profile of horses housed in a loose box and paddock during the different seasons (spring, summer, autumn, and winter). Blood samples were performed every 4 h for 48 consecutive hours. Red blood cells (RBCs), hemoglobin (HGB), hematocrit (HCT), white blood cells (WBCs), platelets (PLTs), and leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes) were analyzed, and, at the same time, environmental conditions were recorded. A statistically significant effect of housing conditions ( p < 0.0001) was observed on all hematological values except for WBC during winter and for neutrophils ( p < 0.0001) during spring and autumn. A statistically significant effect of season ( p < 0.0001) was found for RBC, HCT, and PLT and for all leukocyte cells ( p < 0.0001) except for basophils. The single Cosinor method revealed a daily rhythm of hematological parameters during spring in both groups, and a daily rhythm for lymphocytes and neutrophils was observed during spring and summer in horses kept in a loose box and during winter in horses housed in a paddock. Our results revealed that the response of the immune system is regulated by circadian physiology. Knowledge of the periodic temporal structure of mammals should be considered when evaluating animals' adaptation to temporizations imposed by the environment.

This study aimed to investigate the immune-metabolic status and growth performance of Simmental calves born from cows subjected to pegbovigrastim administration 7 days before calving. Eight calves born from cows subjected to pegbovigrastim administration (PEG group) and 9 calves born from untreated cows (CTR group) were used. Growth measurements and blood samples were collected from birth to 60 d of age. The PEG group had lower body weight from 28 up to 60 d of age (P < 0.01), lower heart girth (P < 0.05), lower weekly and total average daily gain values (P < 0.05) than the CTR group throughout the monitoring period. A decrease in milk replacer (MR) intake was observed in the PEG group compared with the CTR group around 20-28 d of age (P < 0.01). The PEG group had lower values of γ-glutamyl transferase (GGT) at d 1 of age (P < 0.05), Zn at 21 and 28 d of age (P < 0.05), hemoglobin, MCH and MCHC at 54 and 60 d of age (P < 0.01), and higher urea concentration at 21 and 28 d of age (P < 0.05) compared with the CTR group. Lower values of retinol (P < 0.05), tocopherol (P < 0.01), mean myeloperoxidase index (P < 0.05) and higher total reactive oxygen metabolites (P < 0.05) and myeloperoxidase (P < 0.05) were also detected in the PEG group. In light of the results gathered in the current study, it can be speculated that activation of the cow's immune system by pegbovigrastim could have influenced the immune competence, growth performance as well as the balance between oxidant and antioxidant indices of the newborn calf., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objective: The exposure to constant light during the hospitalization was investigated to verify its influence on the daily rhythmicity of Schirmer tear test I (STT I) in the Felis catus., Animal Studied: One group (HG-hospitalized group) was consisted of 10 owned-cats hospitalized to perform a sterilization procedure and was exposed to a 24/0 light/dark (L/D) cycle; the cat control group (CG) was consisted of 10 staff-owned cats living in an indoor environment (12/12 L/D cycle)., Procedure: The STT I values was performed at 4 h intervals over a 48 h period (starting at 8:00 a.m. on Day 1 and finishing at 8:00 a.m. on Day 3) on the left (LE) and right (RE) eyes into two cat groups. A 35 x 5 mm commercial tear test strip was used to record tear production in millimeters wetting per minute (mm/min)., Results: Multivariate for repeated measure analysis of variance (ANOVA) showed a statistically significant effect of time. No difference was found between LE and RE tear production, and between the two experimental conditions. Robust daily rhythmicity was exhibited by the STT I in both eyes during the entire monitoring period in control cats and only during the Day 1 in hospitalized cats., Conclusions: These data are a starting point for evaluating the imbalance of ocular physiology observed in hospitalized cats. Further studies on larger sample size and exposing the animals to various hospitalization procedures are needed to establish whether these alterations are caused by hospitalization procedures or by the light/dark schedules., (© 2022 American College of Veterinary Ophthalmologists.)

This study had a purpose to evaluate the seasonal biodistribution of some trace elements (cadmium, Cd; Lead, Pb; chrome, Cr; and mercury, Hg) in different tissues (muscle, gills, liver, stomach, and intestine) of striped mullet Mugil cephalus (Linnaeus, 1758) and the hematological and biochemical responses of this species to aquatic pollution by trace metals. For this purpose, 80 M. cephalus (20 for each season) were captured in three different stations of Faro Lake, Messina, Sicily. Biometric indices (weight, fork length, and total length) of each fish were registered. The physico-chemical parameters of the water of the study area were also measured, and the content of trace elements in water and sediment was determined. The hematological (white blood cell, WBC; red blood cell, RBC; hemoglobin concentration, Hb; hematocrit, Hct; mean corpuscular volume, MCV; mean corpuscular hemoglobin, MCH; mean corpuscular hemoglobin concentration, MCHC; thrombocytes count, TC), biochemical parameters (aspartate aminotransferase, AST; alanine aminotransferase, ALT; alkaline phosphatase, ALP; lactate dehydrogenase, LDH), and the seasonal concentration of the trace elements in the different tissues of M. cephalus were assessed. Our results showed a different biodistribution of the trace elements in M. cephalus and significant variations of the blood parameters in the different seasons. This research provides a valid contribute to environmental biomonitoring techniques useful in aquatic pollution control and water management. It also contributes to broadening the studies on the improvement of the health and sustainability of aquatic environments., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A total of 112 dogs (49 males and 63 females) belonging to different breeds (i.e., Boxer, Cirneco dell'Etna, Fonni's Dog, Labrador, Crossbreed, German Shepherd, Pit Bull, Shar-Pei, Yorkshire) were analyzed to compare the serum concentration of serotonin, dopamine, norepinephrine, prolactin, beta-endorphins, thyroxine (T4), triiodothyronine (T3), thyroid-stimulating hormone (TSH), and assess whether these parameters can be correlated with the behavioral phenotype of the investigated breeds. T4 was above or below the threshold in 61% and 14% of dogs, respectively; T3, in contrast, 41% of dogs showed values below the limit, while 26% above it. TSH was within the reference range in 58% of dogs; 94% of the dogs had prolactin in the reference range and only five animals showed values above the limit. For beta-endorphins, 49% of dogs had values above the limit, while 46% had values within the reference range. Serotonin and dopamine values below physiological limits were found in 62% and 70% of dogs, respectively. Finally, 61% of the dogs showed norepinephrine values within the reference range. The study confirmed that the assessment of the serum values of hormones and neurotransmitters in dogs could be useful to better understand the behavioral phenotype of the animal and could be useful for breeders and trainers for the selection of the most suitable subjects for specific tasks.

Housing confinement, adaptation to different light/dark conditions, and social deprivation could modify the amount of total locomotor behavior of calves recommended for their psychophysical health. Total locomotor behavior was recorded by means of an activity data logger every 5 min for 6 consecutive days. To do that eight clinically healthy 30-day-old Holstein calves living in calf boxes under natural photoperiod and environmental conditions were enrolled. ANOVA (analysis of variance) showed a statistical effect of the day of monitoring and animal. In the temporal distribution of the resting-activity frequency, it was observed that the calves presented periods of total locomotor behavior with the existence of two peaks, one between 06:00-07:00 and another between 17:00-18:00, which corresponds to time of food intake. In all animals, a diurnal daily rhythm of total locomotor behavior was observed during time of monitoring. Intrasubject and intersubject variabilities were statistically different in mesor, amplitude, and robustness of rhythm. In conclusion, the total locomotor behavior showed a diurnal daily rhythmicity in 30-day-old calves. The characteristics of rhythm were different from individual to individual and from day to day. The recorded intersubject variability must be taken in consideration during the monitoring of farm animals and justifies the application of the device to each animal, as precision livestock farming suggests.

Stress, inflammatory response, and their relationship were investigated in Simmental cows during the transition period (N = 8; 5 multiparous and 3 primiparous) and in their calves (N = 8; 5 heifers and 3 bulls). From cows, blood was collected at days -21 (±4), 0, +1, +7, and +21 days relative to calving. From calves, blood was collected after birth before colostrum intake (0) and then at 1, 7, and 15 days of age. Cortisol, Interleukin 6 (IL-6) and haptoglobin concentration was assessed by ELISA technique; white blood cells (WBC) were assessed using an ADVIA 2120 Hematology System machine. One-way ANOVA showed an effect of time for all the investigated parameters (P < 0.001) except for lymphocytes in peripartal cows. At calving and 1 d after, cortisol concentration was negatively correlated with levels of IL-6, WBC, and monocytes, whereas levels of IL-6 were positively correlated with WBC, neutrophils, and monocytes count. Cortisol, IL-6, haptoglobin, WBC and all leukocyte populations were affected by the age of neonatal calves (P < 0.001) except for neutrophils. A negative correlation between cortisol and IL-6, neutrophils, monocytes and haptoglobin was found at 15 days of age. A positive correlation between IL-6 and haptoglobin at day 15 of age, and with neutrophils and monocytes at days 7 and 15 of age was found. A positive correlation was obtained between cortisol levels measured in cows around calving and those obtained in calves after birth before colostrum intake (r = 0.83), and between IL-6 concentrations obtained from cows at calving and 1 d after and those obtained in calves at day 1 of age, after the colostrum intake (r = 0.93 and 0.79, respectively). The study suggests that immune function of peripartal cows is in an active state and that, in addition to other well-known factors driving the changes of parameters herein investigated, cortisol could have a role in the immune-modulatory adjustment during peripartum in cows. Furthermore, it can be hypothesized that cortisol is transferred from the cow to newborn calf through the placenta only and not through colostrum, whereas IL-6 levels in calves during the 24 h after birth seem to be influenced by IL-6 values measured in cows around calving due to its transfer through colostrum., Competing Interests: Declaration of competing interest The authors declare no competing interests. None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

This study aimed to investigate the serum concentration of osteocalcin (OC), parathyroid hormone (PTH), calcitonin (CT), calcium (Ca), phosphorus (P), alkaline phosphatase (ALP) and acid phosphatase (AP) in healthy-weaned-foals from 8 months of age until the first year of life. Moreover, the correlation of investigated parameters and foals' age, as well as the relationship between the serum values of PTH and the other markers of bone remodeling were assessed. From 20 foals (10 fillies, 10 colts), blood samples were monthly collected (from 8 to 12 months of age, T1-T5) and the concentration of OC, CT, Ca, P, ALP, AP and PTH was assessed. Two-way repeated measures analysis of variance (ANOVA) showed age-related differences on OC concentration in both fillies (P = 0.008) and colts (P = 0.03) with higher OC values at T5 than T1 and T2. Sex-related effect on OC concentration was found with higher OC levels in colts compared to fillies (P = 0.01). A significant positive correlation between the OC, PTH, CT and APL values and foal's age was found in fillies and colts. PTH levels were positively correlated with the values of OC in fillies and colts. Obtained findings confirmed the dynamic bone turnover during growth in foals. The modification of PTH and OC confirmed the potential role of these parameters as indicator of bone growth and metabolism. The sex-related difference observed in OC concentration opens up new insights into the possible involvement of sex hormones in the regulation of bone metabolism in prepubertal foals., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Phytotherapy has been promoted for the treatment of liver diseases in dogs. The interest in identifying the antioxidant/hepatoprotective potential of various plants is increasing. Every 10 days for 30 days, forty dogs were subjected to blood sampling and hepatic ultrasound assessment. Clinically healthy dogs (group A) and dogs with liver enzyme and ultrasound hepatic aspects and sizes outside the physiological range (group B) were divided into two subgroups. Every day for 30 days, one subgroup received Epacare pet + pasta ® and the other received a placebo. Differences due to groups were observed in dROM, SHp, AST and LDH serum concentrations. The administration of Epacarepet + pasta ® for 30 days had an influence on the serum concentration of dROMs, SHp, AST, ALT, ALP, LDH, and urea. The application of paired Student's t-test showed a decrease in the longitudinal and transverse liver axis size. In conclusion, feed supplementation with Epacare pet + pasta ® had a beneficial effect on the antioxidant status and liver enzymes in animals with liver enzymatic alterations and in healthy dogs., Competing Interests: The authors declare no conflict of interest.

This study evaluated (i) whether weight gain and levels of inflammatory and immune markers including white blood cells (WBC), serum haptoglobin, C-reactive protein, albumin, and globulin fractions change between the Nero Siciliano pig breed and the crossbreed Landrace x Large White (LxLW) reared under the same environmental and farming conditions; and (ii) whether a liquid whey diet supplementation affects the investigated parameters in both genotypes. In this study, 10 crossbreed LxLW and 10 Nero Siciliano pigs were given control feed, representing the control groups (CTRC and CTRNS), whereas 10 crossbreed LxLW and 10 Nero Siciliano pigs were given control feed supplemented with liquid whey for 2 months, representing the experimental groups (WC and WNS). From each pig, body weight and blood were collected before experimental diet supplementation (T0), and one (T1) and two (T2) months after the start of the diet supplemented with whey. The white blood cell count (WBC), serum haptoglobin, C-reactive protein, total proteins, albumin and globulin fraction concentration were assessed. Two-way analysis of variance showed an increasing trend of body weight both in the control and experimental groups of the two pig genotypes throughout the monitoring period ( p < 0.01) without a significant effect of genotype and diet ( p > 0.05). The concentration of haptoglobin, β1- and β2-globulins was affected by pig genotype, diet supplementation, and time ( p < 0.01). The values of WBC, C-reactive protein, albumin, α-globulins, and A/G ratio were affected by diet supplementation ( p < 0.01) and time ( p < 0.01) without an influence of genotype ( p > 0.05). Nero Siciliano pigs showed lower levels of haptoglobin, β1-globulin, and β2-globulin compared to crossbreed LxLW. Nero Siciliano pigs and crossbred LxLW fed with liquid whey showed lower levels of WBC, haptoglobin, C-reactive protein, α-, β1-, and β2-globulins and higher values of albumin compared to control groups. The results reinforced the hypothesis that autochthonous breeds possess higher resilience to farming conditions when compared to allochthonous breeds. Moreover, an immune-modulatory and an anti-inflammatory power of liquid whey dietary supplementation is suggested probably thanks to its content in natural bioactive substances including anti-inflammatory cytokines and anti-oxidative factors., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 D'Alessandro, Arfuso, Floridia, Tardiolo, Fazio, Giannetto, Piccione and Zumbo.)

The aim of this study was to evaluate some physiological parameters as total locomotor activity and subcutaneous temperature in two different bovine breeds under tropical environment. In this study, total locomotor activity and subcutaneous temperature were evaluated in 18 clinically healthy female cows. All cows were divided into 3 groups: group A1 was constituted by 6 Holstein Bos taurus, group A2 was constituted by 6 female Herford Bos taurus cows, and group B was constituted by 6 female Bradford Bos indicus cows. Data recording of total locomotor activity (TLA) and subcutaneous temperature (ST) was performed by means of a subcutaneous thermometer (Thermocron) and an actimeter ACTIWATCH® (Cambridge Neurotechnology Ltd.) based on accelerometer technologies equipped on all subjects to record TLA. At the same time, thermal and hygrometric recording were carried out, considering the subtropical climate of Santa Fe. The application of GLM for statistical analysis showed a significant effect (P < 0.05) on statistical model and time of the day on TLA and ST for all groups; no significative effects on animal parameters were found for ST and TLA except for group B. Circadian parameters have been evaluated according to the single cosinor procedure of ST that showed a diurnal daily rhythmicity for all investigated groups and TLA which is focused almost during the photophase for groups A1 and A2 and during scotophase for group B. Considering different species and breeds, and different environmental conditions, this study suggested that some subjects may be much more able to adapt themselves to environmental stress than others. During their evolution from Bos taurus, zebu cattle (Bos indicus) have acquired genes that confer thermotolerance. Subjects from Bos indicus breeds are better able to regulate body temperature in response to heat stress than Bos taurus subjects., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Lithium (Li + ) represent a substance particularly used for human psychiatric disorders but its therapeutic effect is not well documented for equine specie. For its neurotrophic effect, it may be used as a possible doping substance in horses sports competitions. The purpose of the present study was to determine the different bioaccumulation of lithium concentrations in different biological substrates (blood, serum, mane, and tail), in 30 horses (15 geldings and 15 mares) and hematological parameters as blood biomarkers for lithium bioaccumulation. (RBC, WBC, HGB, HCT, MCV, MCH, MCHC). The lithium concentration in substrates were analyzed by one-way ANOVA. An unpaired t-test was performed between geldings and mares in all substrates. Furthermore, a single regression analysis (Pearson) has been carried out between the lithium concentrations of all biological substrates with each other and between the substrates with the hematological parameters. Our results showed a higher value of lithium concentration in blood and mane in total, and a higher value in mane and tail for mares than geldings. Correlation analysis showed only a significant correlation among blood, serum, and tail. A significates statistical correlation between blood and all hematological parameters except for WBC and MCH was observed. A significant correlation was shown for serum and RBC, HGB and HCT, and between tail and HGB, HCT and MCHC. No significant correlation was observed for mane and hematological parameters. These data could be useful for the evaluation of biodistribution of lithium in an athletic horse in relation to different gender for future use in doping control., (Copyright © 2022. Published by Elsevier Inc.)

Protection against infectious diseases can be obtained with vaccines generating immunogenic response through a combination of humoral and cellular immunity. In this study haematological and serum protein electrophoretic profiles of horses vaccinated against herpesvirus 1 (EHV-1) and 4 (EHV-4) were evaluated. Blood samples were collected from 16 horses before (T0), after 24h, 48h, 72h, 1st week, 2nd week and 3rd week (T1I, T2I, T3I, T7I, T14I and T21I) from the first EHV vaccine-dose administration as well as before (TPRE II ), and after 24h, 48h, 72h, 1st week, 2nd week, 3rd week and 4th week (T1II, T2II, T3II, T7II, T14II, T21II and T28II) from the EHV vaccine-booster. Total leukocyte values increased at T1I, T1II, T3II and T28II compared to T0 (P < .01). Higher lymphocytes and lower neutrophils values were found after first vaccine-dose and vaccine-booster administration compared to the T0 (P < .01). Monocytes showed higher values at T14II than T0 (P < .01). Higher serum values of total proteins, α1-, α-2-, β1-, β2- and γ-globulins were found in horses after first vaccine-dose and vaccine-booster administration (P < .01). Gathered results suggest that horses vaccinated against EHV1 and EHV-4 exhibited a dynamic change of WBC, lymphocytes, neutrophils and monocytes. Moreover, the analysis of serum electrophoresis pattern suggested that EHV vaccination induced the development of inflammation and antibody response in vaccinated horses as highlighted by the increase of α-, β- and γ-globulin fractions. These changes probably reflect the systemic immunological adaptation of animals to EHV vaccine., (Copyright © 2022. Published by Elsevier Inc.)

In clinical practice, an important aspect of animal health status evaluation is the body temperature recording obtained using different methods. Within them, the non-contact and non-invasive infrared thermometer can provide an accurate estimation of body temperature improving the quality of care and medical decision. To evaluate the use of the monitoring of auricular temperature to improve the quality of care and medical decision. To evaluate the use of the monitoring of auricular temperature as indices of body temperature, as well as rectal temperature in eleven clinically healthy mixed bred cats, eleven Rottweiler dogs, and eleven Italian saddle horses temperature values were obtained by means of an infrared thermometer in the left and right ears and by means of a digital thermometer in the rectum. Two-way analysis of variance (ANOVA) indicated a statistically higher rectal temperature value than the temperature recorded in the left and right ears (P<0.001). In all species, no differences between the right and left ear were recorded (p>0.05). Cats and dogs showed a statistically higher auricular (p<0.001) and rectal (p<0.01) temperature compared to horses. No differences were observed between cats and dogs (p>0.05). Auricular and rectal temperature agreement was shown by the Bland-Altman test. Between the two methods, the average difference was 1.6 °C for cats, 1.4 °C for dogs, and 3.3 °C for horses. In dogs, rectal and auricular temperature showed a positive correlation (r 2 =0.78). In conclusion, only in healthy dogs maintained in controlled conditions the monitoring of auricular temperature reflects the clinical practice gold standard of core body temperature measurement represented by rectal temperature. The lower value of about 1.5 °C must be taken into consideration when this technique is used., (Copyright © 2022. Published by Elsevier Ltd.)

Thermoregulation is an important mechanism to ensure thermal homeostasis. In horses, different housing conditions could influence this mechanism by exposing animals to different microclimates. The circadian rhythm of body temperature is well known. For this reason, we wanted to investigate the daily rhythm of cutaneous (jugular, shoulder, inner thigh and croup) temperature in comparison to rectal temperature in athletic horses kept under two different housing conditions. Ten clinically healthy Italian Saddle horses were divided into two equal groups: the first group was housed in individual boxes, and the second group was kept in paddocks. In all horses, rectal and cutaneous temperature measurements were performed every 4 h for 48 consecutive hours during different seasons (spring, summer, autumn and winter), respectively, by means of digital and infrared thermometers. Ambient temperature, relative humidity and ventilation were recorded simultaneously. A general linear model (GLM) was applied to the recorded temperature values displaying a significant effect of season, time of day, site of recording and housing condition (p < 0.001). No statistical differences were found between the two days of monitoring (p = 0.49). A trigonometric statistical model (single cosinor method) was applied to investigate the circadian rhythm of rectal and cutaneous temperatures in the two different groups. Our results showed circadian rhythmicity of rectal temperature during all seasons and in both groups. Cutaneous temperature shows daily rhythmicity that was different in the various regions and was influenced by housing conditions and seasons. Application of GLM also showed a statistically significant effect of season and site of recording (p < 0.0001) on all circadian parameters and of management condition on amplitude and robustness (p < 0.0001). No statistical differences between the two days of monitoring were observed (p = 0.68). These findings are probably due to the animals' thermoregulatory mechanisms ensuring heat exchange between the body and the environment, and that was influenced by the microclimate. In particular, the microclimate influenced the thermodispersion disrupting the daily rhythmicity of some body regions, in horses kept in boxes. Only the temperate climatic conditions observed in summer guarantee the expression of the daily rhythmicity of all body surfaces in both management conditions. These results contribute to the knowledge of the mechanisms of homeostasis and control of body temperature in the athletic horse kept in different housing conditions, with a chronophysiological interpretation that completes the proper management of thermal well-being in horses., (© 2022. The Author(s) under exclusive licence to International Society of Biometeorology.)

The aim of the present study was to investigate the response to physical exercise of some ocular physiological variables (ocular temperature and intraocular pressure) in horses performing a jumping course in the morning (a.m.) or in the afternoon hours (p.m.), before and after the exercise, in correlation with the rectal temperature. Data collection was carried out on 14 clinical healthy Italian saddle horses. All horses were trained from 9:00 to 10:00 a.m. and from 19:00 to 20:00 p.m. according to a specific training program. Ocular temperature (OT), rectal temperature (RT) and intraocular pressure (IOP) were determined. Statistical analysis showed no differences between the results for the left and the right eyes. The application of two-way repeated-measures analysis of variance (ANOVA) showed a statistically significant effect of time (before vs. after) on IOP ( p = 0.0001). RT and OT were statistically influenced by time of day and by experimental conditions (a.m. vs. p.m.) ( p = 0.0001). Bland-Altman (B-A) testing showed an agreement between the values of RT and OT. Our results showed a correlation between increased body temperatures and IOP variations during exercise, so they can probably be considered indicators of athletic horses' physical fitness during exercise.

This study aimed to evaluate the changes in serum oxidant and antioxidant parameters together with the serum values of homocysteine (Hcy) and muscle enzymes including creatine kinase (CK), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in regularly trained athletic horses during official races of 1200, 1600 and 2000 m. Thirty Thoroughbred horses were divided into three groups of 10 subjects each according to the race distance: Group 1, 1200 m race; Group 2, 1600 m race; Group 3, 2000 m race. Blood samples were collected from horses 1 week prior to the race (1WB), on the day of the race at rest (TREST), immediately after the race (TPOST), and after 30 (TPOST30) and 120 (TPOST120) minutes. Serum total proteins, reactive oxygen metabolites (dROMs), thiol antioxidant barrier (SHp), antioxidant barrier (Oxy-ads), Hcy, CK, AST and LDH values were assessed. A two-way repeated measures ANOVA did not show differences referable to the race distance (Group effect) on all investigated parameters (p > 0.05). An exercise effect on oxidative stress markers, Hcy and muscle enzymes herein investigated was found in all groups (p < 0.001). A Pearson’s test showed dROMs positively correlated with SHp, Oxy-ads and Hcy after exercise (p < 0.05). This study suggests that, though well-trained racing horses are subjected to oxidative stress during a race, a proper antioxidant capacity may improve their ability to cope with exercise-induced oxidative stress.

During shearing, animals' welfare is adversely affected and acute stress occurs. Once animal perceives a threat, it develops behavioral, autonomic, endocrine or immune responses to maintain homeostasis. This study aimed to investigate the usefulness of eye temperature assessment by infrared thermography (IRT) to evaluate acute stress response in sheep undergoing a shearing procedure. From each animal, blood sampling, rectal and eye temperature assessment were performed before shearing (T PRE ), 5 (T POST5 ) and 60 (T POST60 ) minutes after the end of shearing procedure. On blood samples the serum cortisol concentrations were evaluated. Rectal temperature (T RECTAL ) was measured using a digital thermometer. Thermographic acquisitions of eye temperature were performed from the eye total area (T EYE ) and from three regions of interest (lateral canthus, T ROI-1 ; central cornea, T ROI-2 ; medial canthus, T ROI-3 ). One-way analysis of variance showed a significant increase of serum cortisol concentration, T RECTAL , T EYE and T ROI-3 (p < 0.001). Serum cortisol was positively correlated with T RECTAL and T ROI-3 at T POST5 and T POST60 . T RECTAL resulted positively correlated with T ROI-3 at T PRE , T POST5 and T POST60 . Agreement between T RECTAL and each eye temperature considered (T EYE , T ROI-1 , T ROI-2 , T ROI-3 ) has been shown by Bland-Altman plots at each time point of monitoring period. The findings obtained in the current survey suggest that the medial canthus is the most suitable region for eye temperature measurement to asses stress response in animals. Moreover, this study highlighted the usefulness of IRT as an immediate and non-invasive physiological measure to assess stress response in sheep., (Copyright © 2022. Published by Elsevier Inc.)

In veterinary medicine, the gold standard for assessing body temperature is rectal temperature assessment. Considering that this procedure is stressful for many species, in particular for cats, it could be clinically important to consider an alternative approach for the monitoring of core body temperature. The aim of this study was to test if cutaneous temperature measurements by means of different infrared thermometers are in agreement with the most commonly used method for body temperature measurement in cats. The cutaneous temperature was recorded in the jugular, shoulder, rib, flank, and inner thigh, using three different non-contact infrared thermometers (IR1, IR2, and IR3) in 20 cats. The cutaneous temperature was then compared to the rectal temperature, recorded by means of a digital thermometer. The obtained data indicated that the cutaneous temperature recorded by the infrared thermometers was not in agreement with the data recorded by the digital thermometer in the rectum. In cats, the use of non-contact infrared thermometers gave no reproducible or constant data to justify their application for the recording of body temperature instead of rectal temperature recording. In addition, the infrared temperature measurement devices generated results that were not in good agreement among themselves, providing a novel result of clinical importance.

Raman spectroscopy is a rapid qualitative and quantitative technique that allows the simultaneous determination of several components in biological fluids. This methodology concerns an alternative technique to distinguish between non-healthy and healthy subjects. Leishmaniasis is a zoonosis of world interest, the most important agent is L. infantum. Dogs are the principal reservoirs affected by a broad spectrum of clinical features. During a clinical exam, blood samples were collected in tubes without anticoagulants, from twenty two dogs. One aliquot was used for serological test for Leishmaniasis, one aliquot was subjected to the Raman spectroscopic analysis. Animals were divided into two groups of equal subjects, Leishmania group (LG) constituted by infected dogs, and control group (CG) constituted by healthy dogs. The acquired spectra were different in the region 1200-1370 cm -1 , in which it is possible to distinguish the amide III vibration (~ 1300 cm -1 ). In LG, an evident shift to the shortwave region is observed in spectral frequencies of the band centered at ~ 1250 cm -1 . Our results distinguished between LD group and CG. Further studies are necessary to exclude the effect of metabolic modification due to disease on the recorded spectra changes and to consolidate the achievability of Raman spectroscopy as rapid and less expensive diagnosis of Leishmaniasis., (© 2022. The Author(s).)

Horse trekking is a non-competitive sport that, as other kinds of exercise, involves skeletal muscle effort and may implicate significant energy expenditure leading to stress. The aim of the current study was to assess the effect of trekking on metabolic and stress response as well as muscle damage markers in clinically healthy horses. Thirty horses were equally divided in 3 groups according to trekking course: Group A (trekking course 15 km long), Group B (trekking course 35 km long starting;), Group C (three trekking courses for three consecutive days; during the first day the trekking course was 25 km long, Group C 1 ; during the second day the trekking course was 40 km long, Group C 2 ; during the third day the trekking course was 25 km long, Group C 3 ). Blood samples were collected from each horse before trekking (TPRE), after 5 min (TPOST) and 1 h from the trekking end to assess lactate, glucose, total proteins, non-esterified fatty acids (NEFAs), creatinine, aspartate aminotransferase (AST), creatine phosphokinase (CPK), lactate dehydrogenase (LDH) and cortisol concentration. Higher cortisol, lactate, glucose, NEFAs, creatinine, CPK, LDH and AST values were found after trekking than rest (P < .001). Horses of Groups C 2 and C 3 showed higher values of the investigated haematochemical parameters and muscle enzymes than Groups A and B. The findings suggest that trekking induces a metabolic reaction accompanied by an acute stress response in horses and underline that this recreational exercise, if not associated with the adequate recovery period, can increase the probability of muscle damage., (Copyright © 2022. Published by Elsevier Inc.)

The current study aimed to investigate the influence of acute stress by shearing procedures on hematological parameters, serum cortisol concentration and serum protein electrophoretic pattern in Comisana sheep. A total of 20 not pregnant and not lactating adult ewes, aged 3−4 years old and with a mean bodyweight of 55.50 ± 3.50 kg, were enrolled in the study. From each animal, blood samples were collected before shearing (TPRE) and 5 (TPOST5) and 60 (TPOST60) minutes after the end of the shearing procedure in order to assess the values of hematological parameters, serum cortisol, total proteins and protein fractions, including albumin, α-, β1-, β2- and γ-globulins. According to statistical analysis results, albumin values were lower at TPOST60 than TPOST5 (p < 0.01), whereas α- and β2-globulins and the A/G ratio were higher at TPOST60 with respect to TPRE (p < 0.01) and TPOST5 (p < 0.01). A higher serum concentration of cortisol was found at TPOST5 and TPOST60 than TPRE (p < 0.01), and at TPOST60 than TPOST5 (p < 0.01). The serum cortisol values were negatively correlated with the serum values of albumin, β1-globulins and A/G ratio at TPOST60, and positively correlated with α- and β2-globulins at TPOST5 and TPOST60. The decrease in the albumin concentration and the increase in the α- and β2-globulins observed in ewes after shearing with respect to the baseline values suggests an acute phase response in shorn ewes. Additionally, the correlation found between the serum cortisol concentration and the serum protein fractions confirmed the immunomodulatory effect of this hormone, emphasizing the linkage between the endocrine and immune systems during an acute stress condition.

Introduction: The aim of the study was to investigate the relationship between biosecurity as scored on the Italian National Animal Welfare Reference Centre (Centro di Referenza Nazionale per il Benessere Animale - CReNBA) checklist and the prevalence of Mycobacterium avium subsp. paratuberculosis , Chlamydophila abortus and Neospora caninum on dairy farms located in Ragusa, Italy., Material and Methods: The checklist was used to assign an animal welfare score to 31 dairy farms. Twenty-one farms with a moderate score (>33%, <66%) formed group 1, and 10 farms with a high score (>66%) were group 2. Blood samples were collected from all cows on each farm to investigate the titres of antibodies against the relevant pathogens. Two-way analysis of variance was applied to assess differences between the two experimental groups and the Mann-Whitney test was applied to evaluate prevalence differences in the tested parasites between the groups., Results: All tested farms had a score that classified them as either good or excellent. A higher incidence of Neospora caninum was observed in group 1. The incidences of the other two parasites were no different between the two groups., Conclusion: The CReNBA checklist represents an impartial, reproducible, functional and smart instrument based on risk analysis and assigns a farm a mathematical animal welfare score. Among the parasites tested for, only Neospora caninum had prevalence influenced by biosecurity. Our preliminary results highlighted the positive associations between good animal welfare, high levels of biosecurity, and the prevention of the infectious diseases caused by the parasites in our focus, which are common on dairy farms., Competing Interests: Conflict of Interest Conflict of Interest Statement: The authors declare that there is no conflict of interest regarding the publication of this article., (© 2022 L. Perillo et al. published by Sciendo.)

The objective of this study was to evaluate the biomarkers of exposure to boron, nickel, arsenic, and antimony in an industrial region, evaluating the bioaccumulation in biological substrates and the correlation with biomarkers such as hematological parameters. Through indication of the accumulation of some minerals in the horse's biological substrates reflects environmental pollution. Moreover, an additional aim of the study was to show whether these contaminants have an influence on the hematological parameters in horses. Blood, serum, mane, and tail samples from 20 horses from an industrial area were analyzed to determine boron (B), nickel (Ni), arsenic (As), antimony (Sb) concentration. Hematological parameters (red blood cell [RBC], white blood cells [WBC], hemoglobin [Hb], hematocrit [Hct], mean corpuscular volume [MCV], mean corpuscular hemoglobin [MCH], mean corpuscular hemoglobin concentration [MCHC], platelet [PLT]) as a biomarker of blood in relation to the bioaccumulation of these elements were analyzed also. Descriptive statistical analysis was performed and single regression analysis (Pearson) and multiple regression analysis (p < 0.05) between blood factors, As, B, Ni, and Sb concentrations, and for each mineral in different substrate, respectively. Results showed a significant correlation between tail and mane concentrations with serum and blood for boron concentration (r = -1 p < 0.05). No significant correlation between sample (feed, hay, mane, tail, and water) concentrations and As, Ni, and Sb were found. A significantly negative correlation with blood parameters (r = -1 p < 0.05) was observed in Boron concentration for mane and tail. This suggests that the mane and tail may be a potential means to investigate suspected exposure to excessive levels of trace minerals., (© 2021 Wiley Periodicals LLC.)

Objective: To evaluate the effects of long-term (30-day) oral administration of melatonin on tear production, intraocular pressure (IOP), and concentration of melatonin in the tears and serum of healthy dogs., Animals: 20 healthy sexually intact adult male dogs., Procedures: 10 dogs were given melatonin (0.3 mg/kg, PO, q 24 h, administered in food at 9 am), and 10 dogs were given a placebo. Tear and serum melatonin concentrations, IOP, and tear production (determined with a Schirmer tear test) were recorded before (baseline) and 30 minutes, 3 hours, and 5 hours after administration of melatonin or the placebo on day 1 and 30 minutes after administration of melatonin or the placebo on days 8, 15, and 30., Results: Data collection time had significant effects on tear production, IOP, and tear melatonin concentration but not on serum melatonin concentration. Treatment (melatonin vs placebo) had a significant effect on tear melatonin concentration, but not on tear production, IOP, or serum melatonin concentration; however, tear melatonin concentration was significantly different between groups only 30 minutes after administration on day 1 and not at other times., Clinical Relevance: In healthy dogs, long-term administration of melatonin at a dosage of 0.3 mg/kg, PO, every 24 hours did not have any clinically important effects on tear production, IOP, or serum or tear melatonin concentrations.

The monitoring of endocrine and immunologic markers during exercise is of paramount importance to assess and/or maintain the physical well-being of athletes as well as to optimize the athletic performance. This study aimed to investigate the linkage between acute stress response and immune status in Thoroughbred horses competing in an official 1300-m race. From 10 horses blood was collected 1 week before the day of the race (1W-BEFORE), before (REST) and immediately after the race to assess the cortisol, Interleukin-1 receptor antagonist (IL-1Ra), total proteins, white blood cells (WBC), red blood cells (RBC), haemoglobin (Hb) and haematocrit (Hct) concentration. Higher levels of cortisol, Il-1Ra, WBC and erythrocytes indices after exercise was found than 1W-BEFORE and REST (P < 0.0001). Cortisol concentration was positively correlated with Il-1Ra, WBC, RBC, Hb and Hct. Overall, the findings suggest that submaximal exercise induces an acute stress response and an immune system reaction in athletic horse. Also, the correlation found between cortisol levels and Il-1Ra, WBC and erythrocytes indices open new scenario on the positive role of this hormone on the complex and dynamic physiological adaptation to exercise implemented by the organism to re-establish the homeostatic equilibrium, and, interestingly, to maintain an adequate anti-inflammatory environment after exercise., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Raman spectroscopy is an inelastic light-scattering phenomenon that provides vibrational spectrum that contains information relative to chemical bonds and symmetry of a specific molecule, allowing the quali-quantitative simultaneous determination of several components in the biological fluids. Raman spectroscopy measurement returns a spectrum over a wavenumber range constituted by several bands representing biomarkers according to investigated biological matrices. In literature, it has been reported that at the frequencies inside the (1,300-1,360) cm -1 total lipids, and tryptophan have been identified. On the basis of that, the aim of the present study was to compare the serum concentration of total lipids and tryptophan in horses subjected to a standardized obstacle course, in comparison with the data obtained in the (1,300-1,360) cm -1 band. At this purpose, five clinically healthy and regularly trained Italian Saddle horses aged between 8, and 10 years old performed with the same rider an obstacle course of 350 m/min with twelve 1.30 m high jumps preceded by warm-up. Blood samples were collected by means of jugular venipuncture into a vacutainer tubes with clot activator at rest, after the exercise, and 30 minutes after the end of exercise. A high correlation was observed between the area of total lipids and tryptophan in the (1,300-1,360) cm -1 band and their serum concentrations in all experimental conditions. Our preliminary results give a hint to study the exact correspondence between the area that identify these parameters in Raman spectrum and their serum concentration in athletic horses., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Body temperature reflects the animal health and/or disease conditions. During clinical examination, temperature measurement is a basic step in veterinary medicine. The conventional method used is the rectal thermometry, particularly stressful in some subjects, especially for cats. A less stressful alternative method, such as infrared thermal imaging camera, is used in various fields of medicine for diagnosis, prognosis and correct therapeutic approaches. To evaluate the usefulness of infrared thermal imaging for the assessment of ocular temperature, twenty cats of different breeds (European, Siamese and Persian, 4-6 years old, mean body weight 4.3 ± 0.30 Kg) were enrolled in the study. In order to evaluate the applicability of the ocular temperature assessment through thermal imaging as a tool for measuring the animal's body temperature, the obtained values were compared with the rectal temperature values recorded in each cat by means of a digital thermo-camera. There were no differences between left and right eye; and a difference of about 1.19 °C between the ocular and rectal temperature value was recorded (p < 0.0001). Rectal and ocular temperatures were positively correlated (p < 0.0001; r = 0.93). In conclusion, we show that ocular temperature is an alternative method for body temperature measurement that can be used in clinical evaluation of cats, especially in cases where rectal temperature recording is not possible., (Copyright © 2021. Published by Elsevier Ltd.)

The present study assessed dairy cow welfare through the application of the Italian National Animal Welfare Reference Center (CReNBA) checklist in 36 dairy farms located in Ragusa (Italy) subjected to two different management conditions, housing with free access to pasture (Group 1, farms n = 17) and indoor housing (Group 2, farms n = 19). Five areas of investigation were considered: Area A, "Farm management and personnel"; Area B, "Facilities and equipment"; Area C, "Animal-based measures"; Area D, "Inspection of microclimatic environmental conditions and alarm systems"; and Area E, "Biosecurity". Blood samples were collected by coccygeal venipuncture from all animals (4081 cows). The specific antibodies against Mycobacterium avium subsp. paratuberculosis, Chlamydiophila abortus , Neospora caninum , bovine viral diarrhea virus, and the bovine herpesvirus were assessed by enzyme linked immunosorbent assay (ELISA) serological test. Group 1 (access to pasture) showed a lower value of percentage score recorded in Area A ( p = 0.02) and E ( p = 0.01) than Group 2 (indoor housing). Herpesvirus (Infectious bovine rhinotracheitis - IBR - detection of gB antibodies/IBR-gB) blood concentrations were higher in the cows housed indoor versus those with access to pasture ( p = 0.01). Farm management and personnel (score A) was correlated with the level of bovine viral diarrhea virus (τ = 0.3754) and bovine-herpesvirus-specific antibodies (IBR-gB) (τ = 0.4159). "Biosecurity" percentage score showed a significant correlation with Chlamydiophila abortus (τ = -0.4621) in the cows with access to pasture and IBR-gB (τ = 0.3435) in the cows housed fully indoors. Group 2 showed a significantly reduced level of antibodies against Neospora caninum . In conclusion, differences in the welfare assessment score were observed in the "Farm management and personnel" and "Biosecurity" between the two management conditions. It had an effect on the prevalence of herpesvirus, which occurred more in cattle with access to pasture. Therefore, an accurate application of the checklist could be an instrument to prevent and control the spread of infections in farms.

A deep knowledge on metabolic response of athlete horse to exercise is of paramount importance for the development of optimal training programs to attain equilibrium between good performance and animal welfare. This study investigated the dynamic change of metabolic, clotting times and peripheral indices of central fatigue in horses competing in an official 44 km endurance race.  From 6 horses blood samples were collected at rest (T PRE ), within 5 minutes from the end of race (T PE5 ), after 30, 60 minutes after the end of race (T PE30 and T PE60 ) to investigate the values of blood lactate, glucose, platelets (PLTs), serum leucine, isoleucine, valine, tryptophan, the ratio between tryptophan and branched-chain amino acids (BCAAs) levels (Try/BCAAs), total proteins, non-esterified fatty acids, plasma thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time and fibrinogen (Fb). Lactate, PLTs, non-esterified fatty acids, BCAAs and Try/BCAAs showed higher values at T PE5, T PE30 and T PE60 than T PRE (P < .0001); glucose showed lower values at T PE5, T PE30 and T PE60 than T PRE (P < .0001). Higher TT values were found at T PE5, T PE30 and T PE60 than T PRE, and at T PE5 than T PE30 and T PE60 (P < .0001); Fb was higher at T PE5 than T PRE, T  PE30 and T PE60 (P < .0001). This survey confirms that horses performing prolonged physical exercise experienced glycolysis, lipid mobilization and amino acid metabolism to provide substrates as fuel for muscle contraction., (Copyright © 2021. Published by Elsevier Inc.)

The purpose of the current study was to investigate the effects of two commonly used sedation protocols in dogs, acepromazine (ACP) and acepromazine-methadone (ACP-MET) combination, on tear production measured by the Schirmer Tear Test (STT) 1. We hypothesized that both sedation protocols cause a reduction in canine tear production for a variable time. Fifteen client-owned dogs were recruited for the study. Each dog was subjected to sedation twice, 2-3 weeks apart, and they were randomly allocated to one of two groups receiving ACP (0.015 mg kg -1 ) or ACP-MET (0.010 mg kg -1 and 0.2 mg kg -1 ) intramuscularly. In both eyes, tear production was measured 15 min before sedation (T0) and 20 min (T20 m), 40 min (T40 m), 1 h (T1), 2 h (T2), 4 h (T4) and 8 h (T8), after drug administration. Two-way repeated measures ANOVA, followed by the Bonferroni post hoc test ( p < 0.05), showed a significant effect of time ( p < 0.0001) and treatment ( p < 0.0001). A significant decrease in tear production at T20 m, T40 m, T1 and T2 compared to T0 was observed in the ACP experimental protocol, while in the ACP + MET protocol, this reduction persisted until T8. Comparing the two experimental protocols, no statistically significant differences were observed at T0 or T20 m, and STT 1 values were statistically lower in the ACP + MET than the ACP protocol at the other data points. In the ACP + MET group, at T40 m, 100% of dogs showed STT 1 readings lower than 15 mm/min. This finding is clinically relevant as it can predispose dogs to corneal injuries. The major reduction in tear production due to the ACP + MET protocol proves the need for adequate corneal hydration, particularly to discourage its use in animals with altered tear production. The data obtained provide important information helping clinicians to better manage the drug's effects on tear production.

An innate 24 h circadian clock drives various behavioral processes via expression of clock genes that regulate circadian rhythmicity and temporal signals. Elucidating the gene expression in goats may contribute to improving the knowledge of the regulation of circadian rhythms in this species. Five nonpregnant and nonlactating Maltese goats with no evidence of disease were kept in an indoor pen under the natural long photoperiod (05:05-20:56 h) and natural environmental temperature (23°C and 60% RH). They were fed an Alfalfa hay and concentrate mixture provided twice a day; water was available ad libitum. Blood samples were collected every 4 h over a 48 h period into PAX gene Blood RNA Tubes and stored at -80°C until processing. Clock genes ( Clock; Cry1; Cry2; Per2; Per3 ) were determined using real-time quantitative polymerase chain reaction. During the experimental period, locomotor activity was monitored by an actigraphy-based data logger that records a digitally integrated measure of motor activity as a means to assess indices of discomfort during study and stability of the circadian rhythm. All of the tested genes showed daily rhythmicity in their expression in whole blood. Differences in their circadian parameters were observed. Mesor and amplitude were statistically different among the tested gene (Mesor: F (4.30)  = 205.30; p < .0001; amplitude: F (4.30)  = 104.80; p < .0001), with each gene showing its acrophase at a different time of day (F (4.30)  = 81.17; p < .0001), and differences were observed between the two days of monitoring (F (1.30)  = 10.25; p = .003). The application of two-way analysis of variance (ANOVA) on robustness of rhythm values did not show statistical differences among the tested genes (F (4.30)  = 1.83; p = .14) and between the two days of monitoring (F (1.30)  = 1.16; p = .28). Locomotor activity data recording were in accordance with the data reported in literature, indicating the absence of discomfort or alteration of circadian rhythms during the experimental period. Our results support the presence of a cyclic transcription of clock genes in whole blood of healthy goats housed under a long light natural photoperiod and natural environmental conditions.

This study aimed to investigate the dynamic change of adrenocorticotrophic hormone (ACTH), cortisol, leptin, mitochondrial uncoupling protein 1 (UCP1), lipids and lipoproteins in mares during late pregnancy and the postpartum period. A total of 20 mares (10 pregnant mares, monitored from 14 ± 2 days before expected foaling until 14 days after foaling, Group A; 10 non-pregnant and non-lactating mares, Group B) were enrolled in the study. Body Condition Score (BCS) and body weight (BW) values were recorded from each animal. In Group A, blood samples were collected on days 14 ± 2 and 7 ± 2 before foaling (T -14 ; T -7 ), and on days 7 and 14 after foaling (T +7 ; T +14 ). From mares of Group B, blood samples were collected at the beginning of the study. The levels of ACTH, cortisol, leptin, UCP1, non-esterified fatty acids (NEFAs), total cholesterol, high-density lipoproteins (HDLs), low-density lipoproteins (LDLs), triglycerides and very-low-density lipoproteins (VLDLs) were investigated. While BCS showed no statistical change throughout the monitoring period ( p > 0.05), all the other studied parameters displayed statistically significant variations in Group A over the peripartum period ( p < 0.0001). A significant effect of pregnancy was found on all studied parameters ( p < 0.001). The ACTH and cortisol levels measured in mares belonged to Group A showed a significant positive correlation with the values of leptin, LDLs, triglycerides and VLDLs, whereas they were negatively correlated with the serum UCP1 and NEFAs values. Together, the findings gathered in this study highlight a dynamic change of serum leptin, UCP1 and lipid parameters in peripartum mares and suggest an interaction of the HPA axis with lipid metabolism and mobilization in mares during the peripartum period in order to deal with metabolic and energy demand and maintain energy homeostasis.