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The study aimed to evaluate how plasma total proteins, their fractions (i.e. albumin, α1-, β-, γ-globulins) and A/G ratio change in lambs and kids over the first month of life. Ten female Maltese kids and 10 female Comisana lambs were enrolled in the study. Starting from the first day after birth, and for a period of 4 weeks, blood samples were taken from each subject. Specifically, for each kid and lamb, blood sampling was performed within 30 min after birth (T0) (pre-colostral), after colostrum intake (T1), and, weekly, for the subsequent 28 days (T7, T14, T21 and T28). The application of two-way analysis of variance for repeated measures (ANOVA) showed the effect of time in the kids and lambs for the values of plasma total proteins, albumin, α1-, β-, γ-globulins and A/G ratio (p

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.

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.

Abstract 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

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.

The evaluation of thermoregulation in homeothermic animals is important for their health assessment. Body temperature is influenced by the circadian rhythm, which, through certain signals, is regulated by the suprachiasmatic nucleus. Temperature is collected by various methods; to reduce the invasiveness of rectal temperature sampling, considered the most accurate, infrared thermography has been used. The aim of the present review was to describe the circadian variability of average body temperature in several domestic animal species. In addition to variations due to the circadian rhythm, a number of conditions that can influence body temperature have also been studied. One example of this is exercise, which occurs in the case of domestic animals such as horses, dogs, and donkeys. In particular, it has been analysed in athletic horses, where, following intense exercise, the circadian rhythm of temperature is altered. The daily temperature variation during pregnancy and the neonatal period was also analysed. The circadian rhythm of temperature is influenced by the gestational period of sheep and goats, but this is affected differently depending on the type of species. The same is true for the neonatal period, in kids, lambs, and cattle, where the circadian rhythm is established at different times.

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.

Horsehairs present several common characteristics in their chemical composition and molecular structure. The present study aims to analyze the physical characteristics of horsehairs belonging to different breeds. Morphological analysis of the horsehair fibers was performed using Scanning Electron Microscopy (SEM) and molecular structural characterization using the Raman Spectroscopy (RS) technique. Horse hairs were collected from three different horse groups (group A: mixed-breed; group B: Italian saddle; group C: thoroughbred). Each group was constituted of five horses with a mean body weight of 475 ± 25 kg, aged between 12 and 15 years old. SEM images showed differences in the surface layer (cuticula) and diameter size of horsehairs referred to different breeds. The investigation conducted through RS showed differences in the S – O band, located at 1044 cm−1, where cysteic acid is one of the amino acid constituents of α-keratin; in CH2 bending mode and CH stretching, located at ~1450 cm-1 and ~2900 cm-1, respectively. These differences could be attributed to genetic predisposition or metabolism; they could represent the real differentiation among the breeds, detectable by using RS.

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.

This study aimed to assess the effect of exercise on serum electrophoretic protein pattern, C-reactive protein (CRP) and platelet aggregation in horses subjected to a jumping exercise. The possible relationship between acute-phase reactions and platelet reactivity in the context of exercise was investigated. Blood samples were collected from 10 jumper horses at rest (TREST), within 5 min from the end of exercise (TPE5), and 30 min (TPE30) and 60 min after exercise (TPE60). The serum values of total proteins; CRP; albumin; α1-, α-2, β1-, β2- and γ-globulins; and the maximum degree of aggregation and the initial velocity of aggregation (slope) were evaluated. According to one-way analysis of variance, CRP and α1-, α-2, β1- and β2-globulins increased after exercise compared with rest condition (p < 0.001), whereas albumin and platelet aggregation showed lower values after exercise than at rest (p < 0.001). CRP and α1-globulin values were negatively correlated with both platelet aggregation indices at TPE5, whereas no significant correlation among these parameters was found at TREST, TPE30 and TPE60. This study provides evidence that an acute-phase response occurred in horses after the jumping exercise and suggests a linkage between the inflammatory status and the platelet responsiveness in horses during exercise.

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.

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-, β-, 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; β-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.

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.

This study aimed to investigate the effects of diazinon on fish, focusing on hepatotoxic biomarkers and the potential protective effects of silymarin supplementation. One hundred eighty rainbow trout were randomly assigned to four groups: control, diazinon exposed (0.1 mg L−1), silymarin supplemented (400 mg kg−1), and diazinon + silymarin. Blood samples and liver tissue were collected after 7, 14, and 21 days of exposure to analyze biochemical parameters and oxidative biomarkers. Diazinon exposure in fish resulted in liver damage, as indicated by increased antioxidant enzyme activities in the hepatocytes. Silymarin showed the potential to mitigate this damage by reducing oxidative stress and restoring enzyme activities. Nevertheless, diazinon increased creatine phosphokinase activity, which may not be normalized by silymarin. Exposure to diazinon increased glucose, triglyceride, and cholesterol levels, whereas total protein, albumin, and globulin levels were significantly decreased in fish. However, silymarin controlled and maintained these levels within the normal range. Diazinon increased creatinine, urea, uric acid, and ammonia contents. Silymarin could regulate creatinine, urea, and uric acid levels while having limited effectiveness on ammonia excretion. Furthermore, diazinon increased malondialdehyde in hepatocytes, whereas administration of silymarin could restore normal malondialdehyde levels. Overall, silymarin showed potential as a therapeutic treatment for mitigating oxidative damage induced by diazinon in fish, but its effectiveness on creatine phosphokinase, glutathione reductase, and ammonia may be limited.

The current study compared the faecal microbiota composition of two pig breeds (autochthonous vs. commercial) to understand what happens after the integration of liquid whey in the diet and what the role of the host genetic is. The trial was conducted for 60 days, and the faecal microbiota composition was investigated at three time points, T0, T1 (after 30 days) and T2 (after 60 days) in 30 female pigs (20 commercial crossbred and 10 Nero Siciliano pigs). The animals were divided into four groups (two control and two treatment groups). Generally, in both breeds, Firmicutes (51%) and Bacteroidota (36%) were the most abundant phylum whereas Prevotella, Treponema and Lactobacillus were the most abundant genera. The two breeds have a different reaction to a liquid whey diet. In fact, as shown by PERMANOVA analysis, the liquid whey significantly (p < 0.001) affects the microbiota composition of crossbreeds while not having an effect on the microbiota of the Nero Siciliano. Despite this, in both breeds Bifidobacterium and Ruminococcus have been positively influenced by liquid whey and they promote intestinal health, improve immunity, increase performance, and feed efficiency. In conclusion, the integration of liquid whey had a different effect on the Nero Siciliano and crossbred pig breeds, emphasizing the importance of the host genetic profile in determining the faecal bacterial composition.

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.

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 assessed the potential effect of a co-feed liquid whey-integrated diet on the fecal microbiota of 14 crossbred pigs. The experimental design was as follows: seven pigs were in the control group, fed with a control feed, and seven were in the experimental group, fed with the same control feed supplemented daily with liquid whey. The collection of fecal samples was conducted on each animal before the dietary treatment (T0) and one (T1), and two (T2) months after the beginning of the co-feed integration. In addition, blood samples were collected from each pig at the same time points in order to evaluate the physiological parameters. Taxonomic analysis showed a bacterial community dominated by Firmicutes, Bacteroidetes, Spirochaetes, and Proteobacteria phyla that populated the crossbred pig feces. The diversity metrics suggested that the co-feed supplementation affected some alpha diversity indexes of the fecal microbiota. In addition, the differential abundance analysis at the genus level revealed significant differences for various genera, suggesting that the liquid whey supplementation potentially influenced a part of the bacterial community over time. Spearman’s correlations revealed that the differential abundant genera identified are positively or negatively correlated with the physiological parameters.

Blood cells are constantly exposed to several pollutants, including xenobiotics, and they can be considered a useful target for pollution exposition of the animal. The aim of this study was to investigate the effects of two xenobiotics (Quaternium-15, a preservative used in personal care products, and Thiacloprid, a neonicotinoid pesticide) on the haematological profile and erythrocyte osmotic fragility (EOF) of equine and bovine blood samples. Ten blood samples from horses and cows were exposed for 24 h to Quaternium-15 at a concentration of 0.1 and 1 mg/L and to Thiacloprid at a concentration of 30 and 60 µg/mL. A decrease in the values of the red blood cells, white blood cells, haematocrit, haemoglobin, mean corpuscular volume, and platelets, and an increase of EOF were found in blood samples exposed to xenobiotics compared to the control. According to the results gathered in the current study, the two xenobiotic compounds herein tested negatively affect the haematological indices causing haemolysis both in cattle and horse blood. This study, despite being preliminary, emphasizes the concept that blood cells are an excellent target for evaluating the effects of xenobiotics.

The expression of miRNAs is one of the main epigenetic mechanisms responsible for the regulation of gene expression in mammals, and in cancer, miRNAs participate by regulating the expression of protein-coding cancer-associated genes. In canine mammary tumors (CMTs), the ESR1 gene encodes for ERα, and represents a major target gene for miR-18a and miR-18b, previously found to be overexpressed in mammary carcinomas. A loss in ERα expression in CMTs is commonly associated with poor prognosis, and it is noteworthy that the downregulation of the ESR1 would appear to be more epigenetic than genetic in nature. In this study, the expression of ESR1 mRNA in formalin-fixed, paraffin-embedded (FFPE) canine mammary tumors (CMTs) was evaluated and compared with the expression levels of miR18a and miR18b, both assessed via RT-qPCR. Furthermore, the possible correlation between the miRNA expression data and the immunohistochemical prognostic factors (ERα immunoexpression; Ki67 proliferative index) was explored. A total of twenty-six FFPE mammary samples were used, including 22 CMTs (7 benign; 15 malignant) and four control samples (three normal mammary glands and one case of lobular hyperplasia). The obtained results demonstrate that miR-18a and miR-18b are upregulated in malignant CMTs, negatively correlating with the expression of target ESR1 mRNA. Of note, the upregulation of miRNAs strictly reflects the progressive loss of ERα immunoexpression and increased tumor cell proliferation as measured using the Ki67 index. The results suggest a central role of miR-18a and miR-18b in the pathophysiology of canine mammary tumors as potential epigenetic mechanisms involved in ERα downregulation. Moreover, as miRNA expression reflects ERα protein status and a high proliferative index, miR-18a and miR-18b may represent promising biomarkers with prognostic value. More detailed investigations on a larger number of cases are needed to better understand the influence of these miRNAs in canine mammary tumors.

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.

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.

Abstract Background Leishmania infection in cats is being increasingly reported in endemic areas. Nevertheless, only a few clinical cases have been described in cats, and even fewer have provided information on the response to treatment and a proper follow-up. Here we report a case of feline leishmaniosis not associated with any other disease or co-infection and document its response to allopurinol treatment and long-term follow-up data. Results A 6-year-old domestic shorthair female cat was referred for nodular blepharitis, mucocutaneous ulcerative lesions of the mouth and lymph node enlargement. The cat was moderately anaemic, hyperglobulinaemic and tested negative for feline leukaemia virus and feline immunodeficiency virus. Fine needle aspirates of nodules and mucocutaneous lesions showed the presence of numerous amastigote forms of Leishmania. Leishmania infection was further confirmed by serology (IFAT test, 1:640) and real-time PCR (RT-PCR) on blood and conjunctival swabs. The cat was treated with allopurinol (20 mg/kg SID), which was clinically effective, although the cat remained Leishmania-positive in serology and RT-PCR on blood and conjunctival swabs. Allopurinol treatment was interrupted after seven months because of the healing of all lesions and lack of compliance by the owner. After two years, the cat relapsed displaying almost the same clinical signs and clinicopathological alterations. On this occasion, the parasite was isolated by culture and identified as belonging to L. infantum. Allopurinol treatment was started again but was interrupted several times because of the itching side effect observed. The cat worsened progressively and died two months after the relapse without any chance to shift the treatment to another molecule (e.g. meglumineantimoniate or miltefosine). Conclusions Out of all documented cases of feline leishmanosis, the present case has the longest follow-up period and it is one of the few in which the parasite was isolated and identified. It further confirms the potential progression of Leishmania infection to disease in cats even in the absence of comorbidities. Veterinarians practicing in endemic areas should be aware of this susceptibility, properly include feline leishmaniosis in the differential diagnosis and propose preventative measures to those cats at risk.

MicroRNAs (miRNAs) can act as oncogenes or oncosuppressor genes, and their involvement in nearly all cancer-associated processes makes these small molecules promising diagnostic and prognostic biomarkers in cancer, as well as specific targets for cancer therapy. This study aimed to investigate the expression of 7 miRNAs (miR-18a, miR-18b, miR-22, miR-124, miR-145, miR-21, miR-146b) in formalin-fixed, paraffin-embedded canine mammary tumors (CMTs) by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Twenty-six mammary samples were selected, including 22 CMTs (7 benign; 15 malignant) and 4 control samples (3 normal mammary gland and 1 case of lobular hyperplasia). Oncogenic miR-18a, miR-18b and miR-21 were significantly upregulated in malignant tumors compared with control tissues (p < 0.05). Conversely, oncosuppressor miR-146b was significantly downregulated in benign and malignant mammary tumors compared with control samples (p < 0.05) while, no group-related differences in the expression levels of miR-22, miR-124 and miR-145 were found (p > 0.05). Upregulated miRNAs found here, may regulate genes involved in receptor-mediated carcinogenesis and proteoglycan remodeling in cancer; while miRNA with reduced expression can regulate genes involved in Toll-like receptor and MAPK signaling pathways. According to the results obtained in the current study, the oncogenic and oncosuppressor miRNAs analyzed here are dysregulated in CMTs and the dysregulation of miRNA targets may lead to specific altered cellular processes and key pathways involved in carcinogenesis. Of note, since oncogenic miRNAs predicted to regulate neoplastic cell proliferation and hormonal activities, they may play an active role in neoplastic transformation and/or progression, having mechanistic and prognostic relevance in CMTs.

According to FAO reports, the global mule population counts about 9 million mules. This hybrid cross of a male donkey and a female horse is mainly used for draft purposes because they are thought to be strong and hardy animals. Most consider mules to be less susceptible to disease and fatigue compared to horses. Therefore, the aim of this study was to investigate the effects of fieldwork on biochemical variables and serum amyloid A in working mules. Blood samples were collected from 10 healthy, female, working mules before and after 8 h of fieldwork. According to statistical analysis, a significant influence (p < 0.05) of fieldwork was found on mules’ electrolyte profile with increased levels of sodium, chloride, and calcium, as well as on blood urea nitrogen and creatinine. After a day of fieldwork, serum sodium, chloride, calcium, urea, and creatinine concentrations were increased, supporting decreases in body water and renal blood flow. However, without comparison to a group of mules that were not exercised yet maintained under similar ambient conditions, it is uncertain whether these changes can be attributed to exercise. Further, no change in SAA concentration was found after exercise, indicating that the work performed did not result in systemic inflammation.

Background: Heavy metals are toxic, non-biodegradable substances able to enter the food chain of cows and then transfer to their milk. This study investigated the relationship between the heavy metal concentrations in serum and milk and the environmental pollutants exposure at two different farms in Ragusa, Italy. Methods: To evaluate the concentrations of aluminum (Al), chromium (Cr), iron (Fe), copper (Cu), zinc (Zn), arsenic (As), cadmium (Cd), and lead (Pb), milk and serum samples were collected from 40 Friesian dairy cows from farm 1 at about 3.7 km from an industrial area (group 1) and 40 Friesian dairy cows from farm 2 at about 400 mt from a greenhouse and 6.0 km from a chemical fertilizer factory (group 2). Results: The concentrations of heavy metals measured in serum and milk showed no statistically significant differences between group 1 and group 2. No significant correlation between heavy metals in serum and milk in group 1 was observed. A positive correlation between Zn concentrations measured in milk and serum samples was observed in group 2 (r = 0.35, p = 0.03). Conclusions: The determination of heavy metals in serum and milk can be an important tool to detect the exposure to environmental pollutants and in monitoring the hygienic state of the environment in which milk is produced.

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.

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.

Weaning plays a key role in health status and future performance of calves. The aim of this study was to investigate the effects of weaning age (Wa), early (45 d, EW) or conventional (60 d, CW), on growth performance and metabolic profile of ten Simmental calves (5 EW and 5 CW calves). Daily intake of milk and calf starter was recorded. Blood samples and measurements of body weight (BW), heart girth (HG), and wither height (WH) were collected at −25, −15, 0, 6, and 20 days relative to weaning. Growth performances (BW, HG, WH) were affected by Wa, resulting lower in EW calves compared with CW calves (p < 0.05). Average daily gain was affected by overall Wa and Time but also by the interaction Wa × Time (p < 0.05). EW calves had lower paraoxonase and higher oxidation protein products levels, lower glucose levels in the post-weaning period, lower Ca and cholesterol levels at 20 d after weaning, and higher GGT activity at −25 d from weaning (p < 0.05). A significant interaction effect between Wa and Time was reached for glucose, Ca, cholesterol. In conclusion, weaning Simmental calves at approximately six weeks of age might not affect inflammatory status and liver functionality after weaning. As secondary outcome, even though the low number of animals could represent a limitation, the average daily gain obtained by Simmental calves weaned at 45 d supported this strategy (despite the lower body weight at weaning and after was due only to the age difference of 15 days). Hence, in order to reduce rearing costs, early weaning for Simmental calves (dual-purpose breed, milk and beef) might not jeopardize calf development, as long as calves can reach body gains as reported in the present study.

The increasing levels of anthelmintic resistance together with the restrictions in the use of drugs in food producing animals have enforced the search for sustainable alternative approaches for parasite control. The current study aimed to investigate the safety and the efficacy of a commercially available phytotherapic formulation against gastrointestinal strongyles in donkeys. Twenty-two Ragusana jennies (2.6 ± 0.5 years old) were assigned to two equal groups. One group was treated with two doses of a phytotherapic supplement Paraxitebio® containing Cardus mariano, Eucalyptus globulus, Gentiana lutea, Urtica urens, and Mallotus philippinensis, 14 days apart (Group A). One group was used as negative control (Group B). Individual fecal samples were collected at the beginning of the study (T−1), and after 7, 14, and 28 days (T7, T14, T28). Blood samples were collected on T−1 and T28 in order to assess changes in donkeys' hematological profile. After the initial rise in EPG values observed on T7, Group A showed a significant EPG decrease with lower eggs per gram (EPG) count compared to Group B on T28 and an overall fecal egg count reduction of 56.9% on the same time-point. Hematological parameters were within the normal physiological ranges for enrolled donkeys. However, significant differences in the values of RBCs, Hb, MCHC, MCV, WBCs, eosinophils, and basophils were recorded between groups after phytotherapic treatments, with Group A showing a general improvement in the hemogram picture. The phytotherapic supplement used in the current study was helpful in controlling intestinal parasites allowing a significant reduction in the fecal egg count 28 days after treatment. Further studies are needed to better explore the specific mode of action of the plant-derived formulation herein tested as well as to encourage their use as tool for the control of equine strongylosis under multimodal integrated approach in dairy donkey farms.

Electric impedance spectroscopy techniques have been widely employed to study basic biological processes, and recently explored to estimate postmortem interval (PMI). However, the most-relevant parameter to approximate PMI has not been recognized so far. This study investigated electrical conductivity changes in muscle of 18 sea bass specimens, maintained at different room temperatures (15.0 °C; 20.0 °C; 25.0 °C), during a 24 h postmortem period using an oscilloscope coupled with a signal generator, as innovative technology. The root mean square (RMS) was selected among all measured parameters, and recorded every 15 min for 24 h after death. The RMS(t) time series for each animal were collected and statistically analyzed using MATLAB®. A similar trend in RMS values was observed in all animals over the 24 h study period. After a short period, during which the RMS signal decreased, an increasing trend of the signal was recorded for all fish until it reached a peak. Subsequently, the RMS value gradually decreased over time. A strong linear correlation was observed among the time series, confirming that the above time-behaviour holds for all animals. The time at which maximum value is reached strongly depended on the room temperature during the experiments, ranging from 6 h in fish kept at 25.0 °C to 14 h in animals kept at 15.0 °C. The use of the oscilloscope has proven to be a promising technology in the study of electrical muscle properties during the early postmortem interval, with the advantage of being a fast, non-destructive, and inexpensive method, although more studies will be needed to validate this technology before moving to real-time field investigations.

Background: In this study, the clinical response to treatment with intravenous lipid emulsion (ILE) for nine cats intoxicated with permethrin has been described. Methods: The enrolled cats showed acute onset of seizures, tremors and hypersalivation that were partially controlled with the administration of benzodiazepines and intravenous fluid therapy. Due to worsening clinical signs, intravenous lipid emulsion (intralipid 20%; 1.5 mL/kg in bolus, followed by 15 mL/kg for 1 h in infusion) was administered. Results: After ILE treatment, all cats recovered rapidly and were discharged within 24–54 h. Intravenous lipid emulsion appears to be a relatively safe and inexpensive alternative, with a short hospitalization time. Conclusions: Intravenous lipid emulsion could be an interesting therapeutic option in emergency and critical care protocols for permethrin toxicosis.

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.

Abstract Background Safety and efficacy of the combined monthly use of spot-on fipronil 6.76% w/v / permethrin 50.48% w/v (Frontline Tri-Act®) and chewable tablets of afoxolaner 1.9% w/w / milbemycin oxime 0.4% w/w (NexGard Spectra®) in dogs was evaluated in a field study over a period of 6 months. Methods Forty-one healthy dogs living in highly endemic area for canine leishmaniosis and other canine vector borne diseases (VBD) were included in the study at the beginning of the Leishmania transmission season. Sixteen dogs were pet dogs living each in a single household; twenty-five dogs were hunting dogs living in three kennels. At inclusion, the dogs were ELISA (rapid test) negative for antibodies to Anaplasma, Borrelia, Ehrlichia, and for antigens of Dirofilaria. The dogs were also negative for blood microfilariae at the Knott’s test, and no clinical or haematological abnormalities were observed. Of the included dogs, six hunting, apparently healthy, dogs were ELISA (rapid test) positive to Leishmania, and some were naturally infected by gastrointestinal nematodes (58.5%) and/or infested by fleas (58.5%) and ticks (9.8%). All the included dogs were treated at Days 0, 28, 56, 84, 112 and 140, and followed-up for efficacy until the study end (Day 168). Results No adverse events related to the two products, nor skin reactions, general signs, or changes in the haematological profile, were observed during the study. At Day 14, anthelminthic efficacy was 100% for Toxocara canis, Toxascaris leonina and Capillaria aerophila, while few hunting dogs were still shedding eggs of Trichuris vulpis (1/25 hunting dog) and Ancylostomatidae (9/25 hunting dogs). All pet dogs were nematode free at the end of the study. Hunting dogs were free of roundworms and whipworms. Twenty-four hours after the first treatment, 95.8% of the ectoparasite infested dogs were free from fleas and ticks. Ectoparasites were significantly controlled during the 6-month study period, with 100% efficacy on both fleas and ticks from Day 56 to Day 168. Blood and serum samples collected on Day 168 were tested for vector-borne pathogens using same methods of the inclusion and no new seroconversions or circulating blood microfilariae were observed. Conclusions Concomitant use of Frontline Tri-Act® and NexGard Spectra® in dogs for six months was well tolerated. The combination was effective in controlling fleas, ticks, gastro-intestinal nematodes, and neither new seroconversion to the tested vector-borne pathogens nor blood microfilariae were detected in treated dogs at the end of the study.

This study aimed to investigate parasite fauna of E. marginatus from the central Mediterranean Sea between Messina and Syracuse. In the present survey; parasite fauna of dusky grouper was investigated for two main reasons: the economic value of this species and the current lack of studies regarding the capture area. Seventy dusky groupers were caught from May 2018 to February 2020. Forty-seven out of the 70 specimens (67.2%) were infected with one or more parasite species. The most abundant species was Prosorhynchus caudovatus (42.9%), followed by Podocotyle temensis (28.6%), Didymodiclinus sp. (18.6%), Philometra jordanoi (5.7%), Anisakis Type II larvae (5.7%). Higher prevalence of infection of P. jordanoi and Contracaecum sp. was found in warm months (March to September), while P. caudovatus and P. temensis were mostly found during cold months. Weight and total length of E. marginatus were positively correlated with the parasitic load of P. jordanoi and Didymodiclinus sp. The different prevalence of parasite infection found between warm and cold months is probably related to the diet of the dusky grouper; which is characterized by mollusks that are intermediate hosts for parasite species found. None of the parasites found in the present survey is responsible for zoonosis

In this study, the presence of Leishmania DNA and blood feeding sources in phlebotomine sand fly species commonly present in Sicily were investigated. A total of 1,866 female sand flies including 176 blood fed specimens were sampled over two seasons in five selected sites in Sicily (southern Italy). Sergentomyia minuta (n = 1,264) and Phlebotomus perniciousus (n = 594) were the most abundant species at all the sites, while three other species from the genus Phlebotomus (i.e., P. sergenti n = 4, P. perfiliewi n = 3 and P. neglectus n = 1) were only sporadically captured. Twenty-eight out of the 1,866 (1.5%) sand flies tested positive for Leishmania spp. Leishmania tarentolae DNA was identified in 26 specimens of S. minuta, while the DNA of Leishmania donovani complex was detected in a single specimen each of S. minuta and P. perniciosus. Interestingly, seven S. minuta specimens (0.4%) tested positive for reptilian Trypanosoma sp. Blood sources were successfully identified in 108 out of 176 blood fed females. Twenty-seven out of 82 blood sources identified in fed females of P. perniciosus were represented by blood of wild rabbit, S. minuta mainly fed on humans (16/25), while the sole P. sergenti fed specimen took a blood meal on rat. Other vertebrate hosts including horse, goat, pig, dog, chicken, cow, cat and donkey were recognized as blood sources for P. perniciosus and S. minuta, and, surprisingly, no reptilian blood was identified in blood-fed S. minuta specimens. Results of this study agree with the well-known role of P. perniciosus as vector of L. infantum in the western Mediterranean; also, vector feeding preferences herein described support the hypothesis on the involvement of lagomorphs as sylvatic reservoirs of Leishmania. The detection of L. donovani complex in S. minuta, together with the anthropophilic feeding-behaviour herein observed, warrants further research to clarify the capacity of this species in the transmission of pathogens to humans and other animals.

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.

The objective of this preliminary study was to evaluate the levels of selected heavy metals in hair samples of Holstein dairy cows reared on agricultural soils characterized by grassland subjected to anthropogenic impacts. Ninety Holstein-Friesian cows were enrolled in the study and divided into six groups according to farm origin. From each animal, hair samples were collected in order to determine the content of aluminum, chromium, iron, copper, zinc, arsenic, cadmium, and lead. One-way analysis of variance was applied to assess statistically significant differences in the studied heavy metals among the six groups. A significant effect of groups (p < 0.05) on all tested heavy metals was observed. In this study, the low concentration of heavy metals in the hair of the studied animals led us to think that the cows were subjected to low levels of these compounds, preventing them from bioaccumulating. Although the current study provides only preliminary results, it highlights the importance of investigating the concentration of heavy metals in cow hair to improve the health and welfare of both humans and animals.

This study aimed to evaluate the possible relationship among UCP1, body weight, rectal temperature and lipid profile in the horse. Thirty clinically healthy Italian Saddle geldings (6–10 years old) were enrolled after the informed owners’ consent. All horses were blood sampled and their body weight and rectal temperatures were recorded. On the sera obtained after blood centrifugation the concentration of UCP1, total lipids, phospholipids, non-esterified fatty acids (NEFAs), triglycerides, total cholesterol, high density lipoproteins (HDLs), low density lipoproteins (LDLs) and very low density lipoprotein fraction (VLDLs) was evaluated. Pearson’s correlation analysis was applied to assess the possible relationship between serum UCP1 concentration and the values of body weight, rectal temperature and lipid parameters. Serum UCP1 concentration showed no correlation with body weight, rectal temperature, HDLs and LDLs values, whereas it correlated negatively with serum total lipids, phospholipids, NEFAs, total cholesterol, triglycerides and VLDLs values (p < 0.0001). The findings suggest that in the adult horse the role of UCP1 is linked to the lipid metabolism rather than to thermoregulation.

The current study aimed to investigate whether peripheral modulators of serotoninergic function and neurohumoral factors’ changes in athletic horses during an official jumping competition, and to evaluate their relationship with the physical performance of competing horses. From 7 Italian Saddle mares (6–9 years; mean body weight 440 ± 15 kg), performing the same standardized warm-up and jumping course during an official class, heart rate (HR) was monitored throughout the competition. Rectal temperature (RT) measurement, blood lactate and glucose concentration, serum tryptophan, leucine, valine, the tryptophan/branched-chain amino-acids ratio (Try/BCAAs), dopamine, prolactin, and non-esterified fatty acids (NEFAs) were assessed before the exercise event (T0), at the end of the competition stage (5 min ± 10 s following the cessation of the exercise, TPOST5), and 30 min after the end of competition (TPOST30). Highest HR values were recorded during the course and at the outbound (p < 0.0001); blood lactate concentration and RT increased after exercise with respect to the rest condition (p < 0.0001). Lower leucine and valine levels (p < 0.01), and higher tryptophan, Try/BCAAs ratio, and NEFAs values were found at TPOST5 and TPOST30 with respect to T0 (p < 0.0001). A higher prolactin concentration was found at TPOST5 and TPOST30 compared to T0 (p < 0.0001), whereas dopamine showed decreased values after exercise compared to rest (p < 0.0001). Statistically significant correlations among the peripheral indices of serotoninergic function, neurohumoral factors, and athletic performance parameters were found throughout the monitoring period. The findings provide indirect evidence that the serotoninergic system may be involved in fatigue during jumper exercise under a stressful situation, such as competition, in which, in addition to physical effort, athletic horses exhibit more passive behavior.

This study aimed to evaluate the hematological and serum protein electrophoretic profiles of horses after inactivated West Nile virus (WNV) vaccine administration. Blood samples were collected from 10 horses before (T0), after 24 h, 48 h, 72 h, 1 week, 2 weeks and 3 weeks (T1I, T2I, T3I, T4I, T5I and T6I) from the first WNV vaccine-dose administration, before the vaccine-booster (TPREII), and after 24 h, 48 h, 72 h, 1 week, 2 weeks and 3 weeks (T1I I, T2II, T3II, T4II, T5II, T6II) from the WNV vaccine-booster. There was a significant increase in lymphocytes and a decrease in neutrophils after both the first vaccine-dose and vaccine-booster administration compared to the baseline values (p < 0.01). Monocytes showed higher values after 72 h, 1 week and 2 weeks from the vaccine-booster (p < 0.01). Higher serum total protein values were found in horses after both the first vaccine-dose and booster administration (p < 0.05). α1-lobulins increased after the vaccine-booster with the highest levels measured at T4II (p < 0.05); α-2- and β-globulin fractions increased throughout the post-vaccine period compared to the baseline values (p < 0.05); and higher γ-globulin values were found before the vaccine-booster (TPREII) and after 24 h, 72 h and 3 weeks from the vaccine-booster (T1II, T3II and T6II). The findings allow us to conclude that the WNV vaccine used in the current study does not alter the overall hemogram picture of horses although it is associated with modulation of leukocyte populations and the serum protein electrophoretic pattern.

Metabolic dysfunction-associated fatty liver disease (MAFLD) includes several diseases, ranging from simple steatosis to steatohepatitis, fibrosis and cirrhosis. Fish-rich diets are considered helpful in the prevention of MAFLD, and the enzymatic hydrolysis of fish waste has been explored as a means of obtaining high-value protein hydrolysates, which have been proven to exert beneficial bioactivities including anti-obesity and hypocholesterol effects. This study aimed to assess the effect of the administration of protein hydrolysates from anchovy waste (APH) for 12 weeks on attenuated high-fat diet-induced MAFLD in apolipoprotein E-knockout mice (ApoE–/–). Thirty ApoE–/– mice were divided into two groups (n = 15/group) and fed a high-fat diet (HFD), with and without the addition of 10% (w/w) APH. After 12 weeks, serum and hepatic lipid profiles, hepatic enzyme activities, liver histology and immunohistochemistry were analyzed to assess hepatic steatosis, inflammation and fibrosis. Twelve-weeks on a 10% (w/w) APH diet reduces total cholesterol and triglyceride serum levels, hepatic enzyme activity and hepatic triacylglycerol content (p < 0.0001), and results in a reduction in hepatic fat accumulation and macrophage recruitment (p < 0.0001). The results suggest that a 10% APH diet has an anti-obesity effect, with an improvement in lipid metabolism, hepatic steatosis and liver injury as a result of a high-fat diet. Protein hydrolysates from fish waste may represent an efficient nutritional strategy in several diseases, and their use as nutraceuticals is worthy of future investigation.

Post-mortem inspection in slaughterhouses plays a key role in the epidemiological surveillance of infectious diseases, including bTB. This study assessed the prevalence of bTB in cattle in Sicily during two different three-year periods (2010–2012; 2017–2019), using data from meat inspections and histopathological methods. Out of 100,196 cattle, 5221 (5.21%) were diagnosed with tuberculous lesions. Higher prevalence of bTB was recorded during the triennium 2010–2012 (6.74%; n = 3692) compared to the triennium 2017–2019 (3.36%; n = 1529), with a decreasing trend in annual occurrence throughout the study period and a heterogenous proportion of infected cattle among the Sicilian provinces (p < 0.01). Lower rates of infection were recorded in animals aged 84 months (p < 0.0001). Pearson’s chi square analysis revealed a higher localization of lesions in the thoracic cavity (p < 0.0001). Gathered findings show that even though bTB occurrence in cattle was greatly reduced over the years, the disease still remains widespread in Sicily, also posing severe implications for public health.

Pregnancy and lactation are physiological periods that result in an increased metabolic demand that, if not satisfied, could provoke a threat to homeostasis. In this study changes in the values of Non-Esterified Fatty Acids (NEFA) and β-hydroxybutyrate (BHB), glucose, insulin, thyroid hormones, milk composition and yield were investigated in buffaloes during the late pregnancy and early lactation. From a total of 50 buffaloes, blood samples were collected -7±5 days before expected calving; +7±5; +30±5 and +50±5 days post-partum; milk samples were collected at the same post-partum time points. On serum samples, the values of Non-Esterified Fatty Acids (NEFA), β-hydroxybutyrate (BHB), glucose, insulin, Triiodothyronine (T3), Thyroxine (T4) and Thyroid-stimulating hormone (TSH) were evaluated. On milk, fat %, protein %, lactose %, somatic cells score (SCS), milk yield and daily milk production (DMP) were assessed. Peripartum period significantly influenced all studied parameters (P

The effect of acute handling stress on haematological profile, blood glucose and lactate (secondary stress markers) in cultured sea bream Sparus aurata was evaluated. Sixty six Sparus aurata were used and equally divided into two groups (A and B). Group A was not subjected to stress, Group B was subjected to acute handling stress. From each fish, biometric data and blood samples were collected to evaluate haematological profile, blood glucose and lactate. Unpaired t-test Student was applied to evaluate possible differences in parameters between the two groups. Red blood cells, haematocrit, haemoglobin, white blood cells (WBC), glucose and lactate showed an increase (P