Data on multiple regression analysis between boron, nickel, arsenic, antimony, and biological substrates in horses: The role of hematological biomarkers.

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.

Diurnal variation in rectal and cutaneous temperatures of horses housed under different management conditions.

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.

Simultaneous recording of subcutaneous temperature and total locomotor activity in Bos taurus and Bos indicus raised in a subtropical region of Argentina.

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.

Bovine besnoitiosis in a cattle herd in Sicily: an isolated outbreak or the acknowledgment of an endemicity?

Bovine besnoitiosis is a debilitating infectious disease caused by Besnoitia besnoiti (Apicomplexa; Sarcocystidae). The disease is mainly characterized by cutaneous and systemic signs, infertility in bulls, and abortion in cows. The current study describes an autochthonous outbreak of bovine besnoitiosis in Sicily, Southern Italy, being the first report of B. besnoiti infection in the island so far. In a cattle farm located in Syracuse province, a 4-year-old Belgian blue bull born in Sicily displayed typical clinical signs of the sub-acute/chronic disease phase with thickening of the skin of the scrotum and testicular degeneration. Histological examination of scrotal biopsies revealed the presence of several tissue cysts of B. besnoiti. The serological analysis of the herd using a commercial ELISA revealed a high seroprevalence (45 out of 55; 82%) of antibodies against B. besnoiti. Few seropositive animals (5 out of 45; 11%) showed clinical signs, cysts in vestibulum vaginae (1 out of 31; 3.2%), and testicular degeneration (4 out of 14; 28.6%) assessed by ultrasonographic investigations. The paucity of clinical signs associated with the high seroprevalence in the farm led to hypothesize that bovine besnoitiosis is endemic in the area though further studies are needed. Local practitioners should be more aware of the disease to facilitate the early detection of cases, prevent the spread of infection, and avoid economic losses and animal health problem.

Infrared methodologies for the assessment of skin temperature daily rhythm in two domestic mammalian species.

To assess the accuracy of infrared methodologies for daily rhythm monitoring of skin temperature, five clinically healthy Italian Saddle gelding horses, and five not pregnant and not lactating Camosciata goats, were monitored every 4 h over a 48 h period. The horses were housed in individual boxes, while the goats in two indoor pens, under natural photoperiod and natural environmental temperature. In each animal, skin temperature was recorded with the use of a digital infrared camera and a non-contact infrared thermometer, in five regions: neck, shoulder, ribs, flank and croup. Recorded values were compared with the well-established daily rhythm of rectal temperature. Rectal temperature was recorded at the same time by means of a digital thermometer. In horses, a lower value of skin temperature was recorded using the infrared thermometer for the croup region compared to shoulder and flank; a lower value of skin temperature was recorded using thermography for the croup region compared to the shoulder. In goats, a lower value of skin temperature was recorded using the infrared thermometer for the croup region compared to the flank. In both species, higher values of rectal temperature were observed, compared to the temperature recorded at the skin regions using the other two methodologies. Cosinor rhythmometry showed a daily rhythm of rectal and skin temperature recorded using both methodologies in all the examined regions. General linear model (GLM) showed statistically significant effect of breed on all rhythmic parameters; of day of monitoring on amplitude; of site of recording (rectal vs skin regions) on mesor, amplitude and acrophase; and no effect of methodologies used. The results of this study show the differences in rhythmicity of various body regions temperature and their differences in comparison with daily rhythm rectal temperature. The use of infrared methodologies was inaccurate in assessing body core temperature, but its use could be considered for the evaluation of inflammation in the different body sites.

Serum haptoglobin and protein electrophoretic fraction modifications in buffaloes (Bubalus bubalis) around calving and during early lactation.

Serum protein distribution and concentration can be affected by different physiological and pathological conditions. The aim of this study was to evaluate the changes in the concentration of serum protein fractions and haptoglobin in clinically healthy dairy buffaloes during late pregnancy and early lactation. Blood and milk samples were collected from 30 buffaloes at around 7 d before expected calving (blood only) and 7, 30 and 50 d after calving. In serum samples, the total protein, haptoglobin, albumin, α1-, α2-, ß1-, ß2-, γ-globulins, and albumin/globulin ratio (A/G) values were evaluated. In milk, fat%, protein%, lactose%, somatic cell score (SCS) were assessed, along with milk yield (MY) and daily milk production (DMP). The peripartum period significantly influenced (P < 0.005) total protein, albumin, haptoglobin, α2-, ß2- and γ-globulins (P < 0.005). Milk yield, DMP and fat% changed significantly throughout the monitoring period (P < 0.005). Milk yield and DMP were positively correlated with total protein, albumin, ß2-globulins and A/G ratio, and negatively correlated with haptoglobin and α2-globulins. These results provide new knowledge about the serum protein electrophoretic pattern in Italian Mediterranean Buffaloes during the last phase of pregnancy and early stages of lactation.

Monitoring changes in body surface temperature associated with treadmill exercise in dogs by use of infrared methodology.

The aim of this study was to evaluate the influence of moderate treadmill exercise session on body surface and core temperature in dog measured by means of two infrared instruments. Ten Jack Russell Terrier/Miniature Pinscher mixed-breed dogs were subjected to 15min of walking, 10min of trotting and 10min of gallop. At every step, body surface temperature (Tsurface) was measured on seven regions (neck, shoulder, ribs, flank, back, internal thigh and eye) using two different methods, a digital infrared camera (ThermaCam P25) and a non-contact infrared thermometer (Infrared Thermometer THM010-VT001). Rectal temperature (Trectal) and blood samples were collected before (T0) and after exercise (T3). Blood samples were tested for red blood cell (RBC), hemoglobin concentration (Hb) and hematocrit (Hct). A significant effect of exercise in all body surface regions was found, as measured by both infrared methods. The temperature obtained in the eye and the thigh area were higher with respect to the other studied regions throughout the experimental period (P<0.0001). RBC, Hb, Hct and Trectal values were higher at T3 (P<0.05). Statistically significant higher temperature values measured by infrared thermometer was found in neck, shoulder, ribs, flank, back regions respect to the values obtained by digital infrared camera (P<0.0001). The results obtained in this study showed that both internal and surface temperatures are influenced by physical exercise probably due to muscle activity and changes in blood flow in dogs. Both infrared instruments used in this study have proven to be useful in detecting surface temperature variations of specific body regions, however factors including type and color of animal hair coat must be taken into account in the interpretation of data obtained by thermography methodology.

Age-related changes of serum mitochondrial uncoupling 1, rumen and rectal temperature in goats.

Thermoregulatory processes are induced not only by exposure to cold or heat but also by a variety of physiological situations including age, fasting and food intake that result in changes in body temperature. The aim of the present study was to evaluate the differences in serum mitochondrial uncoupling protein 1 (UCP1), rumen temperature (TRUMEN) and rectal temperature (TRECTAL) values between adult and kids goats. Ten adult male Maltese goats aged 3-5 years old (Group A) and 30 male kids, raised for meat, were enrolled in this study. The kids were equally divided into 3 groups according to their age: Group B included kids aged 3 months, Group C included kids aged 4 months and Group D included kids aged 5 months. Blood samples and measurements of TRUMEN and TRECTAL were obtained from each animal. One-way repeated measures analysis of variance (ANOVA) was applied to evaluate the effect of age on the studied parameters. Statistically significant higher serum UCP1 levels (P<0.001) were found in Group A as compared to Groups B, C and D. Higher TRUMEN values (P<0.001) were found in Group A than in Groups B, C and D, and in Group B than in Groups C and D. Group A showed lower TRECTAL values (P<0.001) than Groups B, C and D. The Pearson's Correlation test was applied to assess significant relationship among studied parameters showing a statistically significant negative correlation between the values of TRECTAL and serum UCP1 in all studied Groups (P<0.001). These results indicate that goats have good control of body temperature suggesting that further details about the thermogenic capacity and the function of UCP1 in kids and adult goats are worth exploring.

Application of a combined global positioning and heart rate monitoring system in jumper horses during an official competition - A preliminary study.

The aim of this study was to investigate whether a combined global positioning system (GPS)/heart rate (HR) monitoring system is a valuable tool to assess, step by step, the physiological response of HR and its relationship with speed in healthy horses competing in an official show jumping class. Six mares performing a standardised warm-up and jumping course were monitored using a HR/GPS device. Venous blood lactate (BL), assessed before and after exercise, showed a significant increase (P = 0.0004) following the physical effort. One-way analysis of variance (ANOVA) showed significant changes in HR throughout the experimental period. The analysis of HR data recorded during the warm-up jumping stage showed significantly higher HR (P = 0.001) in the recovery period compared to the related jumping phase. Shifting the fence height from 100 cm to 125 cm during the warm-up jumps was also found to cause a significant increase (P = 0.016) in HR. According to these preliminary results, the simultaneous logging of heart rate and speed has the potential to be a reliable and powerful technique for field testing that can help in the monitoring of the horse's response to jumping effort during training and competition.

An In-Depth Look at Fonni's Dog Behavior under Different Outdoor Conditions.

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.

Haematochemical Profile of Healthy Dogs Seropositive for Single or Multiple Vector-Borne Pathogens.

BACKGROUND: the present study aimed to investigate the immunological response to common vector-borne pathogens and to evaluate their impact on haematochemical parameters in owned dogs. METHODS: Blood samples were collected from 400 clinically healthy dogs living in an endemic area (Sardinia Island, Italy). All dogs were serologically tested for VBDs and divided into groups based on their negative (Neg) or positive response towards Ehrlichia (Ehrl), Rickettsia (Rick), Leishmania (Leish), Borrelia (Borr), Anaplasma (Anapl), and Bartonella (Barto). A Kruskall-Wallis's test, followed by Dunn's post hoc comparison test, was applied to determine the statistical effect of negativity and single or multiple positivity on the studied parameters. RESULTS: the group of dogs simultaneously presenting antibodies towards Leishmania, Ehrlichia, and Rickettsia showed higher values of total proteins, globulins, creatine phosphokinase, aspartate aminotransferase, alanine aminotransferase, and amylase than dogs that tested negative or dogs with antibodies toward a single pathogen investigated herein. CONCLUSIONS: Our results seem to suggest that exposure to more vector-borne pathogens could lead to greater liver function impairment and a greater inflammatory state. Further investigations are needed in order to better clarify how co-infections affect haematochemical patterns in dogs living in endemic areas of VBDs.

Automated Hematological Approach and Protein Electrophoretic Pattern in Tilapia (Oreochromis niloticus): An Innovative and Experimental Model for Aquaculture.

This study aimed to assess the usefulness of two innovative automated methods (automated blood count counters and flow cytometry) for hematological investigation in Tilapia to make a contribution to the clinical diagnostics of this farmed species. Moreover, serum total proteins and their electrophoretic fractions (prealbumin, albumin, α-, ß-, and γ-fraction), as health condition indicators, were assessed. The analysis of serum total proteins and electrophoretic fraction showed a normal and typical electrophoretic pattern of healthy fish (serum total proteins, 3.70 ± 0.62 g/dL; prealbumin, 0.44 ± 0.20 g/dL; albumin, 1.17 ± 0.66 g/dL; α-fraction, 1.49 ± 0.64 g/dL; ß-fraction, 0.32 ± 0.16 g/dL; and γ-fraction, 0.29 ± 0.13 g/dL). The relationships between the values of red blood cells (RBCs), white blood cells (WBCs), and thrombocytes (TCs) obtained with the two automated methods were determined using Pearson correlation analysis. The results showed a significant positive correlation between automatic blood cell counting and flow cytometry analysis for RBCs (r = 0.97, p < 0.0001) and WBCs (r = 0.91, p < 0.0001), whereas no correlation was found for TCs (r = -0.11, p = 0.66). The preliminary results gathered in this study seem to highlight the usefulness of the new analytical techniques herein investigated in tilapia, suggesting their application in the hematological investigation of farmed fish species and their usefulness for monitoring the health and well-being of fish reared in aquaculture.

Potential Implications of Acid-Sensing Ion Channels ASIC2 and ASIC4 in Gonadal Differentiation of Dicentrarchus labrax Subjected to Water Temperature Increase during Gonadal Development.

In this study, the expression and implication of acid-sensing ion channels 2 and 4 (ASIC2 and ASIC4) in the gonadal sex differentiation of Dicentrarchus labrax (D. labrax), subjected to increasing water temperatures during gonadal development, were evaluated. Two groups were selected: a control group (CG), in which the average water temperature was maintained at 15 °C and increased to 20 °C in 20 days until weaning; and an experimental group (EG), in which the water temperature was retained at 15 °C for 60 days; thereafter, the temperature was increased daily by 0.5 °C until it reached 20 °C up to the weaning time. Ten fish from the CG and 13 fish from the EG were sampled randomly on the 335th day after hatching (dph). A higher percentage of gonad differentiation in ovaries rather than in testes was observed in the EG compared to the CG (p = 0.01). ASIC2 and ASIC4 were detected for the first time in D. labrax ovaries by indirect immunofluorescence. Both ASIC2 and ASIC4 were expressed in previtellogenic oocytes of ovaries and in scattered cells within some testes, and were most likely intratesticular previtellogenic oocytes in both the CG and EG groups. The CG group showed a higher expression of ASIC4 than the EG cohort (p < 0.05). The results gathered in this study revealed the capacity of water temperature to influence both gonadal differentiation and growth in this gonochoristic fish species, and suggests the possible role of ASIC2 and ASIC4 in gonad differentiation and gamete development in D. labrax.

Using infrared thermography for the evaluation of road transport thermal homeostasis in athletic horse.

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.

Monitoring the physiological inflammatory alertness in horse after road transport.

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-1ß, 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-1ß, 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-1ß, 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.

Diet supplementation with Saccharomyces cerevisiae influences the electrophoretic parameters in blood in young Charolaise bulls.

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-, ß1-, ß2- 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.

Eye Temperature Measured with Infrared Thermography to Assess Stress Responses to Road Transport in Horses.

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.

Persistence of clock gene expression in peripheral blood in dogs maintained under different photoperiod schedules.

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.

Welfare and stress assessment of tourism carriage horses under real working conditions in Sicily.

Animal welfare has become an increasingly important concern regarding equids working as carriage animals. In the present study, the changes in the markers of stress and inflammatory responses as a result of the work performed by tourism carriage horses under real working conditions in Sicily (Italy) were investigated. Twenty-two Standardbreds performed a normal working day in the carriage tourism business during the months of May, June and July 2022, consisting of one day of work for each month. Blood samples were collected in the stables at rest before the tour route (Pre; 07.00 AM) and within 10 min after the end of the workday (Post; 05.00 PM). Haematological parameters, serum concentration of cortisol, total proteins together with the globulin fractions were investigated before and after the carriage work. Environmental temperature, relative humidity and temperature humidity index (THI) were also assessed. The direct erythrocyte indices increased after work compared to rest condition (P < 0.05). The values of cortisol, total proteins and globulins were not affected by carriage work (P > 0.05), while, higher cortisol, total proteins, α1- and α2-globulins values were observed in July compared to May and June (P < 0.05). These changes are probably due to the increase in THI values which showed mild stress in June and high stress in July. This study suggests that the tourism carriage horses herein investigated have adapted to their work activity, however, avoiding working horses during the hottest hours of the day in the summer months is advocated.

Improved quality of life and pain relief in mature horses with osteoarthritis after oral transmucosal cannabidiol oil administration as part of an analgesic regimen.

The aim of this study was to evaluate the effect of oral cannabidiol (CBD) administration in addition to a conventional analgesic protocol on the clinical signs of 20 horses with mild joint osteoarthritis. The horses were randomly assigned to either the control group (C group) or the cannabidiol group (CBD group). Both groups were treated with phenylbutazone for 5 days. The CBD group received 0.03 mg/kg cannabidiol in hemp oil orally once daily for 14 days in addition to phenylbutazone treatment. All subjects were monitored for clinical parameters, oxidative status and blood counts. Pain and quality of life were also assessed using the Horse Chronic Pain Scale (HCPS). The CBD group showed a significant reduction in heart rate, respiratory rate, white blood cell count and oxidative stress (malondialdehyde lipid peroxidation). A significant reduction in HCPS scores was seen in both groups. Lower scores were recorded in the CBD group (3 med; range: 2/4) than in the C group (7 med; range: 4/10). The addition of a cannabidiol-based product to an analgesic protocol was well tolerated and showed positive effects on the treated subjects, improving their quality of life and pain relief.