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The aim of the present study was the evaluation of the bioaccumulation of cadmium (Cd), Zinc (Zn), Chrome (Cr), Lead (Pb), Iron (Fe) and Copper (Cu), in the blood, serum, tail and mane of horses from the industrialized area of Milazzo (Messina, Sicily), to understand the relationships between haematological parameters: Red Blood Cells (RBCs), White Blood Cells (WBCs), Haemoglobin (Hb), Haematocrit (Hct), Mean Corpuscular Volume (MCV), Mean Corpuscular Haemoglobin (MCH), Mean Corpuscular Haemoglobin Concentration (MCHC), Platelets (PLTs) and the concentrations of trace elements. Blood and hair samples from 20 healthy Italian Saddle horses and water, hay and concentrates samples were obtained to determine the haemogram and mineral concentration using a Thermo Scientific iCAP-Q ICP-MS spectrometer. Descriptive analysis showed a higher concentration of Zn, Cr, Pb, Fe, and Cu in the blood and a higher concentration of Cd in the tail than other substrates. A positive correlation was found for Cr (p < 0.0001) and Zn (p < 0.01) between blood and serum substrates, for Zn (p < 0.001) between mane and tail and for Pb (p < 0.01) between blood and mane, while a negative correlation was observed for Cr (p < 0.01) between blood and tail. Results showed a close relationship between the bioaccumulation of certain trace elements in biological substrates and haematological parameters, which represent useful biomarkers suggesting further studies, given the role of haematological parameters in athletic horses.

Over five hundred non-native ant species have spread worldwide, including many that have severe effects on biodiversity, are serious economic pests, or threaten human health and agriculture. The number of species in the Mediterranean is steadily increasing, with Italy being a prominent example. We provide risk screenings for non-native ant species in Italy using a Terrestrial Species Invasiveness Screening Kit using current climate conditions and future predictions. The screened species consist of 15 established and 12 horizon taxa. The results highlight the threat posed by Linepithema humile and Solenopsis invicta, followed by Wasmannia auropunctata, Brachyponera chinensis, and Solenopsis geminata. The threat posed by other tropical invaders such as Anoplolepis gracilipes and Pheidole megacephala depends on climate change scenarios. The Palearctic non-native Lasius neglectus and Tetramorium immigrans species are recognized as intermediate threats, while most screened species are far less threatening. The biology and ecology of most non-native ant species remain scarcely documented. Among the established species, B. chinensis, L. humile, and S. invicta deserve the most attention, while W. auropunctata is rapidly spreading in neighboring countries. Detection is still often accidental and late compared to establishment. Most species first establish around urban areas, making citizen science a promising tool for biosurveillance.

Philometra obladae is a nematode belonging to the family Philometridae. It was morphologically described for the first time in 2008 in Oblada melanura. To date, few data on the molecular characterization of Philometridae have been reported. The aim of the present study was to molecularly characterize Philometra obladae in O. melanura inhabiting the Tyrrhenian coasts off Sicily, Italy. In July 2023, five nematodes were found and morphologically identified as Ph. obladae from the celomic cavity of four O. melanura specimens. Genomic DNA from four nematodes was extracted and two molecular markers, the ribosomal 18S rRNA and the mitochondrial cox1, were amplified using polymerase chain reaction. The sequences obtained were aligned using the MUSCLE algorithm and were used for phylogenetic analyses. Partial sequences of both markers were submitted to GenBank. Phylogenetic trees for both markers resulted in very similar topologies with high posterior probabilities and bootstrap values. Comparisons of our results indicated that Ph. obladae is related to the sequences of other Philometridae isolated from different hosts and different geographic areas. Phylogenetic analysis was carried out to compare the sequences of Ph. obladae with other marine Philometridae, which allowed for the molecular characterization of Ph. obladae as an independent species for the first time.

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.

For their great structural versatility, thiophene-based π-conjugated systems have been widely exploited in the preparation of low band gap materials. Here, we report the synthesis of a highly conjugated tetrathiophene system, namely 1,4-di([2,2′-bithiophen]-3-yl)buta-1,3-diyne (1), that presents two bithiophene units connected at position 3 by a butadiynylene spacer. Single-crystal X-ray diffraction (SC-XRD) analysis elucidated the structure of 1, confirming the planarity of the molecule. The molecule was then electropolymerized onto the surface of a gold-coated piezoelectric quartz crystal, showing a high reactivity that is ascribable to the extended conjugation. The frontier molecular orbital energies of 1 were obtained via DFT optimization performed on the crystal structure-derived molecular geometry. Finally, DFT was also used to estimate the polymer band gap.

The potential of a voltametric E-tongue coupled with a custom data pre-processing stage to improve the performance of machine learning techniques for rapid discrimination of tomato purées between cultivars of different economic value has been investigated. To this aim, a sensor array with screen-printed carbon electrodes modified with gold nanoparticles (GNP), copper nanoparticles (CNP) and bulk gold subsequently modified with poly(3,4-ethylenedioxythiophene) (PEDOT), was developed to acquire data to be transformed by a custom pre-processing pipeline and then processed by a set of commonly used classifiers. The GNP and CNP-modified electrodes, selected based on their sensitivity to soluble monosaccharides, demonstrated good ability in discriminating samples of different cultivars. Among the different data analysis methods tested, Linear Discriminant Analysis (LDA) proved to be particularly suitable, obtaining an average F1 score of 99.26%. The pre-processing stage was beneficial in reducing the number of input features, decreasing the computational cost, i.e., the number of computing operations to be performed, of the entire method and aiding future cost-efficient hardware implementation. These findings proved that coupling the multi-sensing platform featuring properly modified sensors with the custom pre-processing method developed and LDA provided an optimal tradeoff between analytical problem solving and reliable chemical information, as well as accuracy and computational complexity. These results can be preliminary to the design of hardware solutions that could be embedded into low-cost portable devices.

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.

Multiple factors can facilitate invasion success, with the absence of natural enemies, such as predators and parasites, recognised as conferring a significant advantage on invasive over native species. Pumpkinseed Lepomis gibbosus (Centrarchidae) represents one of the most successful freshwater fish invaders in Europe. Previous research has highlighted genetic differences between pumpkinseed populations in Türkiye and those in other European regions, attributed to rapid adaptation to new environmental conditions. This study aimed to investigate whether these highly adapted pumpkinseed populations in Türkiye benefit from a potential release from parasites, as proposed by the enemy-release hypothesis. Genetic characterisation of pumpkinseed populations from both European and Asian parts of Türkiye revealed that they share the same cytochrome c oxidase I haplotype as European populations. Microsatellite analysis indicated low genetic diversity, with STRUCTURE analysis confirming the clustering of all Turkish populations, suggesting a common source. Consistent with the low genetic diversity indicative of a small founding population, we observed a limited number of co-introduced parasite species, including the myxozoan Myxobolus dechtiari, the monogenean Onchocleidus dispar, and the digenean Posthodiplostomum centrarchi. Parasite infection by local parasites acquired in Türkiye was rare. Parasite diversity, species richness, and equitability were low, with only nine parasite taxa identified in all four pumpkinseed populations. The most diverse parasite community was found in Değirmenköy Reservoir, located in the European part of Türkiye, where seven parasite taxa were identified. While our study did not uncover genetically distinct pumpkinseed populations in Türkiye, the fish demonstrated resilience against most local parasite species, potentially providing them with an advantage over native species, aligning with the enemy-release hypothesis.

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.

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

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

Microplastics are transferred to humans through the food chain by consuming food contaminated with microplastics. However, the knowledge about the risks of dietary exposure for humans to these particles is very limited. Moreover, only a few studies on microplastic pollution in fruit and vegetables have been carried on. Thus, this study aims to investigate the presence of microplastics in some of the most consumed fruits and vegetables (pear (Pyrus communis), apple (Malus domestica), tomato (Solanum lycopersicum), onion (Allium cepa), potatoes (Solanum tuberosum), and cucumber (Cucumis sativus)) from Turkey and to evaluate the potential risk for consumers. Fruits and vegetable samples were purchased from different markets and fruiterer (two of each) in Muğla province, Southwest of Turkey. Microplastic extraction processes were carried out on the edible parts of the samples. According to the results obtained, a total of 210 particles (2.9 ± 1.6 particle g−1) were detected in all samples. Any significant difference occurred among the different markets. The maximum average amount of microplastic was determined in tomato samples (3.63 ± 1.39 particle g−1). The highest microplastic intake was with tomato (398,520 particles individual−1 year−1 for Estimated Annual Intake (EAI) and Estimated Daily Intake (EDI) for children 68.24 particles kg−1 day−1). The occurrence of microplastics of big size, that are not allowed to pass by plant xylem transport, suggests that fresh vegetables and fruits can be contaminated with plastic, especially during the production phase, during agricultural activities and during the marketing process (transport to the market and purchasing process).

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.

Mastitis is one of the most considerable and costly diseases for dairy herds, and Staphylococcus spp. is known to be the main causative agent. Although antibiotics are widely used in the treatment of mastitis, this can cause both antibiotic residues in milk and the risk of antibiotic resistance occurrence in bacteria. Thus, in recent years, researchers have focused on alternative treatments for this disease and plants extracts are investigated for this purpose. Pomegranate is widely used as a dye, ornament, and medicinal plants in the industry, and the species has a particularly high economic value in Turkey. This study aims to investigate in vitro the antioxidant and antibacterial properties of the pomegranate flower’s extracts against different Staphylococcus species associated with bovine mastitis. To this aim, pomegranate flowers were collected from different regions in Turkey and extracts were prepared with three different solvents (methanol, ethanol, and water). The retention factor values of the ethanol extract were determined by thin-layer chromatography. The antibacterial activity tests were carried out via the disk diffusion method. In addition, the extracts were tested against the stable DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radicals for antioxidant activity. Four retention factors (0.79, 0.67, 0.58, and 0.33 points) were found for the ethanol extract. The methanol extract showed the highest inhibition zones against coagulase-negative Staphylococcus-37 (CNS-37) and S. aureus-18. The lowest MIC was 6500 μg/mL. The highest antioxidant activity was observed in methanol extracts. As a result, the extracts of pomegranate flowers showed a high antioxidant and antibacterial potential against the examined mastitis pathogens.

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 black soldier fly (BSF), Hermetia illucens, has been recognized as one of the most promising insect species for its ability to valorize organic waste while producing a valuable larval biomass with a great potential as a sustainable source of nutrients, including proteins and bioactive molecules. In the present study, BSF larvae were used to produce and characterize the protein hydrolysates (BPHs) that were then evaluated for their potential biological activity in vitro. The BPHs obtained from the BSF larvae proteins by enzymatic digestion were characterized by Nuclear Magnetic Resonance (NMR) and polyacrylamide gel electrophoresis and assessed for their antioxidant activity (BPHs in the range of 0.1 to 1.5 mg/mL) in L-929 cells. Our findings show that BPHs can exert a dose-dependent cytoprotective role against H2O2-iduced oxidative stress in cells. This antioxidant activity relies on the reduction of ROS levels in challenged cells as measured by flow cytometry and fluorescence microscopy, together with the induction and nuclear translocation of Nrf2, as evaluated by qPCR and indirect immunofluorescence analysis, respectively. Overall, our findings on the remarkable biological activity of the BPHs obtained in a large-scale process strongly suggest the application of BPHs as ingredients promoting animal health in feed formulations.

Mullets (Osteichthyes: Mugilidae) are a euryhaline species widely distributed all over the world, thus representing an excellent study model for host–parasite interactions. From March to June 2022, 150 mullets, belonging to Chelon labrosus (n = 99), Chelon auratus (n = 37), and Oedalechilus labeo (n = 14) species, were caught to identify the helminth parasite fauna of the different mullet species present in the Ganzirri Lagoon (Messina, Sicily, Italy). A parasitological evaluation of the gastrointestinal tract (GIT) was carried out with a total worm count technique (TWC) to detect helminth presence. All collected parasites were stored in 70% ethanol until morphological evaluation, and frozen at −80 °C for subsequent molecular analysis, using 28S, ITS-2, 18S primers. The morphological evaluation allowed for the identification Acanthocephalan parasites (Neoechinorhynchus agilis) from two C. labrosus specimens. Sixty-six samples were positive for adult digenean trematodes (C. labrosus, 49.5 %; C. auratus, 27%, and O. labeo, 50%), molecularly identified as Haploporus benedeni. This study represents the first survey of helminthic parasite fauna of mullets from the south of Italy. The presence of Hydrobia sp. in the stomach contents of mullets allowed us to infer the H. benedeni life cycle in the Ganzirri lagoon.

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.

This study represents the first report of Hysterothylacium incurvum within swordfish (Xiphias gladius) heart chambers. Swordfish is a large pelagic teleost, considered one of the most appreciated fish worldwide. Among swordfish parasites, Anisakis sp. and Hysterothylacium sp. have been used to evaluate biological and ecological aspects of this teleost. Between 2021 and 2022, 364 X. gladius hearts, caught from the Atlantic Ocean (FAO 27.IXa and FAO 34 areas), were collected at the Milan fish market (Lombardy, Italy). Three specimens from FAO 27.IXa was positive for seven adult nematodes (p = 1.55%) within the heart chambers. Of these, three specimens were found within the bulbus arteriosus and 4 in the ventricle. All parasites were stored in 70% ethanol and processed for parasitological and molecular analysis using Cox2, ITS regions/ITS-I-5.8S-ITS-II, and rrnS genes. The analysis allowed us to identify the retrieved parasite as H. incurvum. According to our evaluation, the final localization is due to the movement of L3 larvae from the coelomic cavity to the bloodstream, with consequent development to the adult stage within the heart. Finally, the parasite localization, considered non-marketable fish parts, does not pose a significant risk to consumers, also considering the low zoonotic potential of H. incurvum.

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.

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.

The aim of the study was to determine the potential and sustainable use of pre-commercial product ITTINSECT™ APS V1 as a major protein source in rainbow trout (Oncorhynchus mykiss) diets. A 60-day feeding experiment was conducted to potentially use ITTINSECT as fish meal replacement in the diets of rainbow trout. Five isonitrogenous in dry matter (38% crude protein) and isolipidic (15% crude lipid) diets were produced: a control diet (fishmeal-based) (ITT0) and four experimental diets replacing fishmeal by 25 (ITT25), 50 (ITT50), 75 (ITT75) and 100 (ITT100) %, with ITTINSECT™ APS V1. Triplicate tanks, containing 15 fish each (65.81 ± 1.26 g), were hand-fed to apparent satiation twice every day during the experiment. At the end of the feeding trial, significantly higher growth performance was observed in the group fed ITTM25 and ITTM50 diets. This performance was supported by growth-related gene expressions analyzed in muscle; significantly higher GH and IGF-I genes expression levels were determined in ITT25 and ITT50 when compared to control (ITT0) (p < 0.05). While no significant differences were found between the hematology values (p > 0.05), serum total protein, globulins and glucose levels were significantly different between experimental groups (p < 0.05). In addition to this, the immune-related genes such as TNF-α, IL8 and IL1-β expression levels were determined to be significantly different (p < 0.05). In conclusion, in order to achieve the best growth performance in rainbow trout and enhance sustainable aquaculture practices, replacement of fish meal with up to 50% ITTINSECT™ APS V1 in diets for rainbow trout is suggested.

Innovative and highly performing smart voltammetric immunosensors for rapid and effective serological tests aimed at the determination of SARS-CoV-2 antibodies were developed and validated in human serum matrix. Two immunosensors were developed for the determination of immunoglobulins directed against either the nucleocapsid or the spike viral antigen proteins. The immunosensors were realized using disposable screen-printed electrodes modified with nanostructured materials for the immobilization of the antigens. Fast quantitative detection was achieved, with analysis duration being around 1 h. Signal readout was carried out through a smart, compact and battery-powered potentiostat, based on a Wi-Fi protocol and devised for the Internet of Things (IoT) paradigm. This device is used for the acquisition, storage and sharing of clinical data. Outstanding immunosensors’ sensitivity, specificity and accuracy (100%) were assessed, according to the diagnostic guidelines for epidemiological data. The overall performance of the sensing devices, combined with the portability of the IoT-based device, enables their suitability as a high-throughput diagnostic tool. Both of the immunosensors were validated using clinical human serum specimens from SARS-CoV-2 infected patients, provided by IRCCS Ospedale San Raffaele.

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.

An IoT-WiFi smart and portable electrochemical immunosensor for the quantification of SARS-CoV-2 spike protein was developed with integrated machine learning features. The immunoenzymatic sensor is based on the immobilization of monoclonal antibodies directed at the SARS-CoV-2 S1 subunit on Screen-Printed Electrodes functionalized with gold nanoparticles. The analytical protocol involves a single-step sample incubation. Immunosensor performance was validated in a viral transfer medium which is commonly used for the desorption of nasopharyngeal swabs. Remarkable specificity of the response was demonstrated by testing H1N1 Hemagglutinin from swine-origin influenza A virus and Spike Protein S1 from Middle East respiratory syndrome coronavirus. Machine learning was successfully used for data processing and analysis. Different support vector machine classifiers were evaluated, proving that algorithms affect the classifier accuracy. The test accuracy of the best classification model in terms of true positive/true negative sample classification was 97.3%. In addition, the ML algorithm can be easily integrated into cloud-based portable Wi-Fi devices. Finally, the immunosensor was successfully tested using a third generation replicating incompetent lentiviral vector pseudotyped with SARS-CoV-2 spike glycoprotein, thus proving the applicability of the immunosensor to whole virus detection.

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.

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.

The aim of the present study was to describe gastric granuloma caused by Anisakis pegreffii in Sphyraena viridensis caught in the central Mediterranean Sea. Sixty-eight S. viridensis specimens were collected from different fish markets on the east coast of Sicily. Coelomic organs were observed both macroscopically and with the aid of stereomicroscope. Parasite specimens and lesioned tissues were collected for identification, histological and molecular analyses. Twelve specimens (p = 17.6%) were positive for the presence of nematode larvae, morphologically identified as larvae of Anisakis sp., with values of mean abundance and mean intensity of 0.9 and 4.8, respectively. One large female specimen showed massive parasite infection associated with nodular lesions of the gastric wall. By histology, several nematode larvae encysted through the gastric wall were found. The parasite bodies were surrounded by a granulomatous reaction made up of macrophages, epithelioid cells, some lymphocytes and an external connective sheet. Molecular analysis of 18S rRNA and cox2 genes from Anisakis sp. collected larvae, identified them as A. pegreffii. The lesions here described, though macroscopically superimposable on human eosinophilic granuloma, microscopically showed significant differences in the inflammatory cells involved and in the type of immune reaction.

The ability to make risky decisions in stressful contexts has been largely investigated in experimental settings. We examined this ability during the first months of COVID-19 pandemic, when in Italy people were exposed to a prolonged stress condition, mainly caused by a rigid lockdown. Participants among the general population completed two cognitive tasks, an Iowa Gambling Task (IGT), which measures individual risk/reward decision-making tendencies, and a Go/No-Go task (GNG), to test impulsivity, together with two questionnaires, the Perceived Stress Scale and the Depression, Anxiety and Stress Scales. The Immune Status Questionnaire was additionally administered to explore the impact of the individual health status on decision making. The effect of the questionnaires scores on task performance was examined. The results showed that higher levels of perceived stress and a more self-reported vulnerable immune status were associated, separately, with less risky/more advantageous choices in the IGT in young male participants but with more risky/less advantageous choices in older male participants. These effects were not found in female participants. Impulsivity errors in the GNG were associated with more anxiety symptoms. These findings bring attention to the necessity of taking into account decision-making processes during stressful conditions, especially in the older and more physically vulnerable male population.

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

Pseudorabies virus (PrV) is the etiological agent of Aujeszky’s disease, a viral infection that causes neurological lethal illness in mammals other than swine. Herein, we describe the occurrence of PrV infection in a hunting dog that had been bitten by an infected wild boar in Sicily, reporting for the first time genetic and phylogenetic data on the virus strain isolated in a dog in this Italian region. The dog was referred for severe neurological signs, respiratory distress, and intense itch around the muzzle. Death occurred within 48 h to the onset of clinical signs. On gross examination, self-induced skin lesions to the head due to intense itching and diffuse cerebral congestion were observed, whereas mild, aspecific, nonsuppurative meningitis was histologically diagnosed. Diffuse PrV positivity in neurons of the brainstem was observed by immunohistochemistry. PrV DNA was isolated and amplified from olfactory bulbs by nested PCR, targeting the viral glycoprotein G gene, and the sequence obtained matched with sequences of PrV isolates from dogs and wild boar. Isolation of PrV in the dog herein analysed denotes the spread of the virus in wild boar populations in Sicily and provides a proof of direct interspecies transmission. Thus, there is an urgent need to increase our understanding of the epidemiology of the PrV infection in wildlife to provide tools to trace possible spill over into domestic pigs or other livestock.

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

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.

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.

Pericardial effusion presents clinicians with a challenge when diagnosing the underlying cause and performing a prognosis. Different techniques have been suggested for canine thoracoscopic pericardiectomy with the creation of variable pericardial window size. The aim of this study was to statistically compare the surgical time and achieved window size of the paraxiphoid transdiaphragmatic and monolateral intercostal approaches. The paraxifoid and monolateral intercostal approaches showed a mean surgical time of 55 ± 20.08 (SD) minutes and 13.94 ± 4.61 (SD) minutes, and a mean pericardial window diameter of 4.23 ± 0.80 (SD) cm and 3.31 ± 0.43 (SD) cm, respectively. A significant correlation was observed between the dogs’ bodyweight and window size (r = 0.48; p = 0.04) for both surgical approaches, and between the dogs’ bodyweight and surgical time (r = 0.72; p = 0.0016) for monolateral intercostal approach. All treated dogs showed no clinical signs of recurrent cardiac tamponade during the follow-up. Our results provided useful information to help surgeons make the definitive choice of the surgical technique to treat the pericardial effusion.

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.

The black soldier fly (BSF) Hermetia illucens is receiving increasing attention as a sustainable fishmeal alternative protein source for aquaculture. To date, no studies have explored the effects of fishmeal replacement with BSF V instar larvae or prepupae meals due to their peculiar nutritional properties on fish performances. This study investigated the effects of 100% replacement of fishmeal (control diet) with defatted BSF meals (V instar larvae and prepupae meals, treatments) on growth performance and welfare of zebrafish (Danio rerio), from larvae to adults, in a 60-day feeding trial. Following the inclusion of BSF meals, the expression of key genes involved in growth (igf1, igf2, mstnb, myod1, myog, myf5), hydrolysis of chitin (chia.2, chia.3, chia.5), immune- (il1b, il6, tnfα), and stress- (hsp70 and nr3c1) responses, as assessed by qPCR, was modulated in all of the molecular pathways, except for the stress response. Overall, our findings showed that both BSF meals can totally replace fishmeal without adverse impacts on adult zebrafish growth parameters (final total and standard length, final body weight, weight gain, daily growth rate, specific growth rate) and welfare, with BSF prepupae meal inducing the most beneficial effects, thus suggesting their potential application to meet fish requirements in aquaculture.