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The aim of the present study was to investigate the effects of exercise during canicross competition on urinary biochemical parameters and urinary proteins. Our results showed that, among the parameters analysed in routine urinalysis, pH was affected by exercise and albuminuria also showed changes immediately after physical exercise with a return to baseline within 2 h after the physical activity. In particular, SDS-PAGE electrophoresis allowed the separation of the urinary proteins on the basis of their molecular mass allowing a qualitative evaluation of the urinary proteome and a quantitative analysis of urine albumin and uromodulin, with a significant increase in urinary albumin and a decrease in uromodulin after exercise. In this study, urine was collected by spontaneous voiding and the use of this non-invasive sampling method proved useful in obtaining information on the health of the animals in the immediate moments before and after the competition without increasing the stress of the animals involved in the competition by any visits or invasive manipulations by veterinarians.

Microalgae are used as dietary supplements for humans and animals, due to their excellent nutritional profile. This research aims to characterize Spirugrass®, a co-product obtained after the extraction of phycocyanin from Limnospira platensis, and to evaluate whether a stabilization treatment involving high-pressure processing (HPP) affects its proteomic profile. The research also aims to evaluate the possible use of Spirugrass® as feed integration for honeybee health. Proteins were extracted and fractionated using size exclusion chromatography (SEC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The iron content was measured using atomic absorption spectrometry (AAS). Samples of Spirugrass® were subjected to HPP at 600 MPa for five minutes and the effect on the integrity of the protein profile was analyzed. Finally, nine groups of 30 newly emerged honeybees were supplemented with Spirugrass® in a controlled laboratory experiment. SDS-PAGE and SEC enabled the assessment of the quality and integrity of the Spirugrass® proteome, which contains 80% of the proteins found in the algal biomass, including phycocyanin. The phycocyanin purity indices were 2.07 ± 0.14 and 0.72 ± 0.13 for the L. platensis and Spirugrass® extracts, respectively. Spirugrass® maintains a consistent iron content of 261 ± 15 μg/g dry weight, equivalent to 74% of the iron present in the algal biomass. In both L. platensis and Spirugrass®, iron was predominantly bound to high-molecular-mass proteins, including phycocyanin. Following HPP treatment, differences in the protein profiles were observed, which suggests partial protein degradation. Preliminary data obtained with honeybees are encouraging, as no mortality or adverse effects were observed and Spirugrass® can be considered a promising candidate as feed supplement. Overall, the presence of consistent levels of soluble proteins, as well as protein-bound iron, suggests that Spirugrass® could be used in animal nutrition.

Environment, forage quality, management practices, pathogens, and pesticides influence honeybee responses to stressors. This study proposes an innovative approach to assess colony health and performance using molecular diagnostic tools by correlating hemolymph proteins with common measures of colony strength, prevalent honeybee pathogens (Varroa destructor and Nosema spp.), and essential trace elements (iron, zinc and copper). Colonies were selected from four apiaries located in different environmental and foraging conditions in the province of Bologna (Italy). Hemolymph samples were taken from June to October 2019. The Varroa infestation of the colonies was estimated by assessing the natural mortality of the mites, while the bees were tested for Nosema spp. spores using a microscopic method. Hemolymph proteins were quantified and separated using SDS-PAGE, and colony performance was assessed by determining adult bees, total brood, honey, and pollen reserves. The biomarkers measured proved to be useful for monitoring changes in performance and trophic conditions during summer and early autumn. Significant correlations were found between hemolymph proteins and colony performance measures. A positive correlation between pollen reserves, vitellogenin, and hexamerin 70a highlights the importance of these proteins for successful overwintering. In October, Varroa infestation was negatively correlated with total proteins, vitellogenin, apolipophorin II, transferrin, and hexamerin 70a, with negative implications for overwintering; furthermore, Varroa infestation was also negatively correlated with iron content, potentially affecting iron homeostasis.

Acupuncture has been widely incorporated into rehabilitation protocols for dogs and cats because of its potential analgesic efficacy. The aim of this study was to evaluate the potential positive effect of integrating acupuncture and electroacupuncture techniques on the recovery of ambulation capacity in non-ambulating paraparetic patients undergoing physiotherapy in the postoperative period following mini-/hemilaminectomy for thoracolumbar spinal cord decompression due to acute disc extrusion. Forty-one patients were included and underwent descriptive and analytical statistical analysis, divided into two groups: dogs that received a physiotherapy protocol with acupuncture and dogs that received physiotherapy only. The results showed that the dogs in the acupuncture group had a higher likelihood of regaining ambulation.

Chlorella vulgaris, Arthrospira platensis, Haematoccocus pluvialis, and Phaeodactylum tricornutum are species of interest for commercial purposes due to their valuable nutritional profile. The aim of this study was to investigate the iron content in these four microalgae, with emphasis on their iron bioaccessibility assessed using an in vitro digestion system to simulate the process which takes place in the stomach and small intestine of dogs, followed by iron quantification using atomic absorption spectrometry. Furthermore, the extraction of soluble proteins was carried out and size exclusion chromatography was applied to investigate iron speciation. Significant differences (p < 0.004) in iron content were found between C. vulgaris, which had the highest (1347 ± 93 μg g−1), and H. pluvialis, which had the lowest (216 ± 59 μg g−1) iron content. C. vulgaris, A. platensis, and H. pluvialis showed an iron bioaccessibility of 30, 31, and 30%, respectively, while P. tricornutum showed the lowest bioaccessibility (11%). The four species analysed presented soluble iron mainly bound to proteins with high molecular mass ranging from >75 to 40 kDa. C. vulgaris showed the highest iron content associated with good bioaccessibility; therefore, it could be considered to be an interesting natural source of organic iron in dog nutrition.

Extracts from Boswellia serrata (Bs) and Salix alba (Sa) are used as supplements in poultry feed. The aims of this research were to study the possible effects of dietary supplementation with Bs and Sa extracts on serum and albumen proteins, zinc and iron, and yolk cholesterol content in Leghorn hens during the critical phase of the onset of laying. A total of 120 pullets, 17 weeks of age, were assigned to two groups (control (C) and treated (T), n = 60 each). The T group received a supplement containing Bs (5%) and Sa (5%) for 12 weeks. The study lasted 19 weeks. Serum proteins were fractionated using agarose gel electrophoresis (AGE) and SDS–polyacrylamide gel electrophoresis (SDS–PAGE). Trace elements were determined in serum using atomic absorption spectrometry, and yolk cholesterol was determined using a colorimetric test. No significant differences were observed between control and supplemented hens for the analyzed biochemical indices. Moreover, the supplementation with phytoextracts did not negatively affect the physiological variations in serum proteins; therefore, it can be safely used as a treatment to prevent inflammatory states at onset and during the early laying phase.

IntroductionWe conducted a meta-analysis of quantitative data extracted from selected peer-reviewed papers describing in vivo studies on enteric methane emissions from dairy cattle treated with feed additives, compared with a control group. The aim was to identify feed additives that significantly reduce enteric methane emissions, expressed as g/day, g/kg dry matter intake, g/kg milk produced, and g/kg energy-corrected milk. The feed additives considered were polyunsaturated fatty acids (PUFAs), 3-nitrooxypropanol (3-NOP), essential oils (EO), and monensin. Four electronic databases (PubMed, CAB Abstracts, Web of Science, and Scopus; 2001–2024) were used to retrieve papers, following the PRISMA 2020 statement.MethodsEffect sizes were calculated as log response ratio percentages and analyzed using three-level random-effects models. Heterogeneity, cluster-robust variance estimation, and leave-one-out diagnostics were applied. A total of 34 studies met the inclusion criteria.ResultsOf these, 19 investigated the impact of PUFAs, yielding 45 data points; seven investigated the impact of 3-NOP, yielding 23 data points; eight investigated the impact of essential oils on enteric methane emissions, yielding 12 data points; and three investigated the impact of monensin, yielding four data points. PUFAs significantly reduced enteric methane emissions across all metrics, although high heterogeneity remained (I² ≈ 86%–95%). 3-NOP exhibited the most substantial average reductions in enteric methane emissions; however, the significance of these effects varied depending on the metric and model formulation. In addition, basal crude protein significantly influenced the effectiveness of 3-NOP. The effects of essential oils were generally non-significant and dependent on formulation. Evidence for monensin was limited and descriptive only. DiscussionBased on the current body of evidence, PUFAs and 3-NOP represent the most reliable nutritional strategies for mitigating enteric methane emissions in dairy cows. PUFAs supplementation has been shown to reduce methane production without measurable adverse effects on milk yield or energy-corrected milk. Similarly, 3-NOP produced the greatest average reductions in methane emissions without impairing milk production, although its efficacy may vary depending on diet composition and may decline over time. Future research should prioritize standardized dosing, harmonized measurement methodologies, and extended trial durations that simultaneously assess efficacy, dietary covariates, persistence, productivity, and cost-effectiveness.

Microalgae are a source of bioactive compounds having recently been studied for their possible application as health-promoting ingredients. The aim of the study was to evaluate in an in vitro canine gut model the effects of four microalgae, Arthrospira platensis (AP), Haematococcus pluvialis (HP), Phaeodactylum tricornutum (PT) and Chlorella vulgaris (CV), on some fecal microbial populations and metabolites. The four microalgae were subjected to an in vitro digestion procedure, and subsequently, the digested biomass underwent colonic in vitro fermentation. After 6 h of incubation, PT increased propionate (+36%) and butyrate (+24%), and decreased total BCFA (−47%), isobutyrate (−52%) and isovalerate (−43%) and C. hiranonis (−0.46 log10 copies/75 ng DNA). After 24 h, PT increased propionate (+21%) and isovalerate (+10%), and decreased the abundance of Turicibacter spp. (7.18 vs. 6.69 and 6.56 log10 copies/75 ng DNA for CTRL vs. PT, respectively); moreover, after 24 h, CV decreased C. coccoides (−1.12 log10 copies/75 ng DNA) and Enterococcus spp. (−0.37 log10 copies/75 ng DNA). In conclusion, the microbial saccharolytic activities and the shift in fecal bacterial composition were less pronounced than expected, based on current literature. This study should be considered as a preliminary assessment, and future investigations are required to better understand the role of microalgae in canine nutrition.

This study was conducted to evaluate the safety and the beneficial effects of dietary supplementation with Boswellia serrata (Bs) and Salix alba (Sa) in Leghorn hens during the critical pre-laying and laying phases. A total of 120 pullets, 17 weeks of age, were assigned to two groups (Control—C; Treated—T, n = 60 each). For 12 weeks, the T group received a diet supplemented with 0.3% of dry extracts of Bs (5%) and Sa (5%). The study lasted 19 weeks. Productive performance, serum analytes, H/L ratio, IgA and anti-IBV antibodies were investigated. Water intake was significantly higher, while body and egg weight was significantly lower for the T group (p < 0.05). No other differences were detected in performance parameters, serum analytes, IgA and H/L ratio excluding t0, with a significantly (p < 0.05) higher H/R ratio and higher titers of anti-IBV antibody for the T group. Overall, the data obtained in this study show that the supplementation with Bs and Sa was safe and resulted in an increase in water consumption, a decrease in egg weight, and a sedative effect in the hens. In the future, it would be interesting to test this supplement in hens reared on intensive farms.

Pyrrolizidine alkaloids (PAs) are secondary metabolites produced by plants as a self-defense against insects. After bioactivation in the liver, some PAs can cause acute or chronic toxicity in humans. The aim of this study was to determine the presence of PAs in 121 samples of monofloral and multifloral honey from three different Italian regions (Friuli-Venezia Giulia, Marche and Calabria) to meet the European Food Safety Authority (EFSA) suggestion. An in-house liquid chromatography with tandem mass spectrometry (LC-MS/MS) method was validated according to European Union Reference Laboratory (EURL) performance criteria. This method allowed the detection and quantification of 35 PAs. Of the 121 honey samples, 38 (31%), mostly from Calabria, contained PAs. The total content of the PAs ranged from 0.9 µg/kg to 33.1 µg/kg. In particular, echimidine was the most prevalent PA. A rapid human exposure assessment to PAs in honey and a risk characterization was performed using the EFSA RACE tool. The assessment highlighted a potential health concern only for toddlers who frequently consume elevated quantities of honey. This study showed a low presence of PAs in Italian honey; however, the importance of continuously monitoring these compounds is stressed, along with the suggestion that the relevant authorities establish maximum limits to guarantee support for producers and consumer safety.