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There is a growing interest in developing new molecular markers of heart disease in young dogs affected by myxomatous mitral valve disease. The study aimed to measure 3 circulating microRNAs and their application as potential biomarkers in the plasma of Cavalier King Charles Spaniels with early asymptomatic myxomatous mitral valve disease. The hypothesis is that healthy Cavalier King Charles Spaniels have different microRNA expression profiles than affected dogs in American College of Veterinary Internal Medicine (ACVIM) stage B1. The profiles can differ within the same class among subjects of different ages. This is a prospective cross-sectional study. Thirty-three Cavalier King Charles Spaniels in ACVIM stage B1 were divided into three groups (11 younger than 3 years, 11 older than 3 years and younger than 7 years, and 11 older than 7 years), and 11 healthy (ACVIM stage A) dogs of the same breed were included as the control group. Three circulating microRNAs (miR-1-3p, miR30b-5p, and miR-128-3p) were measured by quantitative real-time PCR using TaqMan® probes. Diagnostic performance was evaluated by calculating the area under the receiver operating curve (AUC). MiR-30b-5p was significantly higher in ACVIM B1 dogs than in ACVIM A subjects, and the area under the receiver operating curve was 0.79. According to the age of dogs, the amount of miR-30b-5p was statistically significantly higher in group B1<3y (2.3 folds, P = 0.034), B1 3-7y (2.2 folds, P = 0.028), and B1>7y (2.7 folds, P = 0.018) than in group A. The area under the receiver operating curves were fair in discriminating between group B1<3y and group A (AUC 0.780), between B1 3-7y and A (AUC 0.78), and good in discriminating between group B1>7y and A (AUC 0.822). Identifying dogs with early asymptomatic myxomatous mitral valve disease through the evaluation of miR-30b-5p represents an intriguing possibility that certainly merits further research. Studies enrolling a larger number of dogs with preclinical stages of myxomatous mitral valve disease are needed to expand further and validate conclusively the preliminary findings from this report.

The epidemiology of Congenital Heart Diseases (CHD.sub.s) has changed over the past twenty years. This study aimed to evaluate the prevalence of CHD.sub.s in the population of dogs recruited in a single referral center (RC); compare the epidemiological features of CHD.sub.s in screened breeds (Boxers) versus non-screened (French and English Bulldogs, German Shepherds); investigate the association of breeds with the prevalence of CHD.sub.s ; determine the popularity and volatility of breeds over a 20-year period; analysed the trends of the most popular breeds in the overall population of new-born dogs registered in the Italian Kennel Club (IKC) from 1st January 1997 to 31st December 2017. The RC's cardiological database was analysed, and 1,779 clinical records were included in a retrospective observation study. Descriptive statistics and frequencies regarding the most representative breeds and CHD.sub.s were generated. A logistic regression model was used to analyse the trends of the most common CHD.sub.s found in single and in cluster of breeds. The relationship between breed popularity and presence of CHD.sub.s was studied. The most common CHD.sub.s were Pulmonic Stenosis, Patent Ductus Arteriosus, Subaortic Stenosis, Ventricular Septal Defect, Aortic Stenosis, Tricuspid Dysplasia, Atrial Septal Defect, Double Chamber Right Ventricle, Mitral Dysplasia, and others less frequent. The most represented pure breeds were Boxer, German Shepherd, French Bulldog, English Bulldog, Maltese, Newfoundland, Rottweiler, Golden Retriever, Chihuahua, and others in lower percentage. Chihuahuas, American Staffordshire Terriers, Border Collies, French Bulldogs, and Cavalier King Charles Spaniel were the most appreciated all of which showed a high value of volatility. This study found evidence for the value of the screening program implemented in Boxers; fashions and trends influence dog owners' choices more than the worries of health problems in a breed. Effective breeding programs are needed in order to control the diffusion of CHD.sub.s without impoverishing the genetic pool.

To evaluate if the functional grading system (Cambridge classification) of brachycephalic obstructive airways syndrome (BOAS) and the temperament score can be useful tools in predicting the feasibility of echocardiographic examination in lateral recumbency. The hypothesis is that the temperament of the dog, rather than the severity of BOAS alone, can exacerbate respiratory symptoms (dyspnea, stertor, stridor and/or cyanosis) during lateral containment. Prospective cross-sectional study. Twenty-nine French Bulldogs were included and classified according to the Cambridge classification for the BOAS and to the Maddern score for the temperament. Receiver operating characteristic analysis was used to evaluate the sensitivity (Se) and specificity (Sp) of the Cambridge classification, of the temperament score and their sum to predict the feasibility of the echocardiography in lateral recumbency without dyspnea/cyanosis. 8 females (27.59%) and 21 (72.41%) males French Bulldogs of 3 years (IQR25-75 1-4), and 12.45 kg (IQR25-7511.5-13.25) were included. The Cambridge classification alone was not predictive for the possibility of performing the echocardiography in lateral recumbency, unlike temperament score and the sum of the two classification indices. The diagnostic accuracy of Cambridge classification (AUC 0.81, Se 50%, Sp 100%), temperament score (AUC 0.73, Se 75%, Sp 69%), and their sum (AUC 0.83, Se 75%, Sp 85%) cut-offs was moderate for each score. The dog's temperament, and therefore its susceptibility to stress, rather than the severity of BOAS (Cambridge classification) alone, is a good predictor of the possibility of performing the echocardiographic examination in standing instead of lateral recumbency.

Specific microRNAs expressions may accurately characterize different stages of canine myxomatous mitral valve disease. This preliminary pilot study aimed to (1) describe the clinical and echocardiographic parameters of Cavalier King Charles Spaniels affected by myxomatous mitral valve disease at different American College of Veterinary Internal Medicine (ACVIM) stages (B1, B2 and C) and healthy control group (ACVIM A), comparing the parameters collected during the first examination (T0) and the end of the follow-up (T1); (2) assess the association between the values of echocardiographic parameters at T1 and the expression profile of miR-30b-5p at T0. Thirty-five Cavalier King Charles Spaniels (median age 4.29 years and median weight 9 Kg) in different ACVIM stages were included (7 A, 19 B1, 6 B2 and 3 C). Inverse probability weighting analysis was performed to estimate the association of the exposure variable (miR-30b-5p) with the outcome variables (clinical and echocardiographic variables). Time was included as variable. The results pointed out that high levels of plasma miR-30b-5p corresponded to lower values of left ventricular end-diastolic diameter normalized for body weight, end-diastolic and end-systolic volumes indexed for body weight, and left atrium-to aortic root ratio. Hence, higher miR-30b-5p expressions were associated with milder forms of mitral valve disease in our study population. In contrast, the results obtained for the intensity of heart murmur, the mitral regurgitation severity, and the Mitral INsufficiency Echocardiographic score) were not statistically significant. A relationship between high abundance of miR-30b-5p and myxomatous mitral valve disease that appear echocardiographically more stable over time has been demonstrated. In conclusion, Cavalier King Charles Spaniels affected by myxomatous mitral valve disease that at the first cardiologic evaluation showed an upregulation of miR-30b-5p are expected to experience lesser variations on their echocardiographic examination between T0 and T1.

Cavalier King Charles Spaniels (CKCS) are predisposed to a hereditable form of myxomatous mitral valve disease (MMVD) with a high incidence and a high risk of disease progression. Gender is recognized as a risk factor for the development of the disease in the breed. However, patient’s morphological traits associated with disease progression have not been identified. The aim of this study was to analyze the association between preclinical MMVD progression and morphometric features in CKCSs. This was a retrospective, time-to-event, longitudinal study. Medical records of 302 CKCSs were reviewed from April 2019 to January 2023. The final sample size counted 52 adult (≥ 1 year) MMVD-affected CKCSs classified as American College of Veterinary Internal Medicine (ACVIM) stage B1, with a minimum 24-month follow-up period and ≥ 2 follow-up examinations. At baseline examination, each dog underwent echocardiography and morphometric evaluation of the body, the head and the thorax; dogs were re-examined at 6- or 12-month intervals, and progression to ACVIM stage B2 was registered as the primary outcome. The median follow-up time was 1270.50 days (interquartile range (IQR, 25 th and 75 th ): 994.75–1525.50 days). In the time-to-event analysis, 17 (32.7%) dogs reached the endpoint, with a median time of 1548 days. Results from the Cox multiple regression analysis with inverse probability weighting (IPW) analysis showed that the following morphometric variables had an association ( p < 0.05) with MMVD progression: thoracic length (hazard ratio (HR): 1.97, p = 0.002), thoracic circumference (HR: 0.61, p = 0.03), and thoracic index (HR: 0.86, p = 0.049). Data obtained from this study suggest that an association between mitral valve disease progression and thoracic morphology exists in MMVD-affected CKCSs classified as ACVIM stage B1. Morphological features should be considered along with other known risk factors for the breed when assessing the risk of MMVD progression in CKCSs.

Non-epidermolytic ichthyosis (NEI) is a hereditary skin disorder affecting several dog breeds, most notably the Golden Retriever. It is primarily caused by a loss-of-function variant in the PNPLA1 gene, while a second, less common form is associated with a deletion in the ABHD5 gene. This retrospective study aimed to assess the prevalence and temporal trends of both mutations in Golden Retrievers tested in Italy between 2017 and September 2025. A total of 508 genetic tests were analyzed, including 463 dogs tested for the PNPLA1 mutation, 42 for the ABHD5 deletion, and 3 for both variants. DNA was extracted from blood or buccal samples and analyzed by real-time PCR followed by confirmatory Sanger sequencing. Among the PNPLA1 tested dogs, 42% were clears (wt/wt), 37% carriers (wt/mut), and 21% affected (mut/mut), with calculated allele frequencies of 60% wild-type and 40% mutant. A significant temporal decline in mutant allele frequency was observed, accompanied by an increasing number of animals tested over time, suggesting growing interest in genetic screening and its impact on selective breeding. Conversely, all dogs tested for the ABHD5 deletion were wild-type, supporting its rarity in the breed. Overall, these findings confirm that PNPLA1 -related ichthyosis remains one of the most prevalent hereditary disorders in Golden Retrievers, although its frequency is decreasing. The results emphasize the effectiveness of genetic testing in disease prevention and highlight the importance of continued monitoring to maintain genetic health within the breed.

Myxomatous mitral valve disease (MMVD) in Cavalier King Charles Spaniels (CKCSs) can be diagnosed at a young age. Early and sensitive biomarkers related to the disease are currently lacking. MicroRNAs (miRNAs) are involved in the onset and progression of canine MMVD. This study aimed to profile the salivary miRNAs associated with MMVD severity in CKCSs and to identify their target genes. This was a pilot prospective, cross-sectional study. Saliva was collected from twenty-five CKCSs belonging to the American College of Veterinary Internal Medicine (ACVIM) stages A (n. 6), B1 (n. 13), and B2 (n. 6), and salivary miRNAs’ expression was profiled by Next Generation Sequencing. miRNAs’ target genes were identified using bioinformatic tools. Results showed that 25 miRNAs were differentially expressed (DE) between ACVIM A and B1 dogs; 35 DE-miRNAs were modulated in ACVIM B2 compared to B1 CKCSs; 4 miRNAs were up-regulated in ACVIM stage B1 patients compared to both ACVIM stages A and B2. These findings confirm that salivary miRNAs can be successfully quantified in CKCSs and that their expression differs across MMVD severity groups. The DE-miRNAs were associated with signalling pathways related to cellular metabolism, survival, and early inflammatory remodeling. Given the exploratory and cross-sectional design, these miRNAs should be considered candidate, non-invasive indicators associated with disease severity rather than established biomarkers. Further longitudinal and validation studies are needed to assess their diagnostic potential.

Taurine deficiency predisposes to the development of nutritional dilated cardiomyopathy and is widespread in dogs fed with non-traditional diets. However, Golden retrievers show lower plasma taurine concentration and an impaired systolic function compared to breeds of the same size and morphotype. For these reasons, it can be difficult to classify a subject from a cardiological point of view, with the risk of considering as pathological characteristics that can be completely normal in this breed. This is a cross-sectional multicenter study. The aims were 1) to identify breed-specific range of serum taurine concentration, 2) to describe a correlation between serum taurine concentration and echocardiographic parameters of systolic function in clinically healthy Golden retrievers fed with traditional diet, 3) to identify a correlation between thyroid hormones, serum taurine concentration and echocardiographic indices. Sixty clinically healthy Golden retrievers (33% males, 67% females) were included. Fifty-three dogs were fed with traditional diets and their range of serum taurine concentration was 398.2 (31.8–430) nmol/ml. Serum taurine concentration was found to be negatively correlated to systolic internal diameter of the left ventricle and systolic and diastolic left ventricular indices and volumes obtained with different methods, whereas was positively correlated to the left ventricle ejection and shortening fractions but difference was not statistically significative. A weak but significant correlation between serum taurine and T4 was demonstrated. Serum taurine median values in dogs with normal systolic function were higher than in dogs with impaired systolic function. A cut-off of serum taurine concentration of 140.6 nmol/ml had a moderate sensitivity and specificity in the identification of an impaired left ventricular systolic function (AUC 0.6, Se 78%, Sp 44%). This study showed that the median serum taurine concentration was significantly lower in dogs with impaired systolic function. Therefore, echocardiographic monitoring is recommended in all dogs with serum taurine concentration lower than 140.6 nmol/ml.

Standard transthoracic echocardiography is considered the non-invasive gold standard for the diagnosis of most cardiac diseases. Defining reproducibility, repeatability, and reliability of this exam is imperative to reduce errors in clinical evaluations. The present study aimed at: (1) evaluating the reproducibility and repeatability of 15 echocardiographic parameters in dogs by analyzing measurements obtained from several operators with different levels of experience and comparing them to the ones obtained from two board-certified operators (gold standards - GSs); (2) assessing whether different formative paths have an influence on the variability of the echocardiographic measurements. Fifty-one operators have been included in this study, along with two diplomates of the European College of Veterinary Internal Medicine - Cardiology. Ten dogs were enrolled, 5 Golden Retrievers and 5 Cavalier King Charles Spaniels. Echocardiographic examination was performed on each dog by one GS and several operators on the same day. Results show the highest deviation from the GS and a poor inter-operator reproducibility for the M-mode measurements of the interventricular septum and the left ventricular free wall. Differently, M-mode-obtained internal diameters of the left ventricle in systole and diastole, and measurements of the aortic annulus and root show moderate to excellent intra- and inter-operator reliability and a good concordance with the GSs, demonstrating that all the operators correctly assess left ventricular systolic function and dilation, and evaluate the aortic valve. Furthermore, a specialist clinical activity, more than the acquired theoretical knowledge, affects the reliability of the echocardiographic examination by reducing the difference from the GS’ measurements.

PURPOSE: The purpose of the study is to compare the performances of two passenger cars: an electric vehicle (EV) and an internal combustion engine vehicle (ICEV) paying particular attention to the production of electricity that will charge the EV. Even if many similar comparative life cycle assessments (LCAs) exist (Nordelöf et al. J Life Cycle Assess 19(11):1866–18990, 2014), only few have focused their attention on evaluating which is the kind of electricity that will recharge EV batteries (Hawkins et al. Int J Life Cycle Assess 17(8):997–1014, 2012). METHODS: Despite its relevance, many EV LCA studies have used a ready-to-use dataset to evaluate the power mix that supplies electricity to EV. The present paper tries to better define the power mix that recharges EV batteries in Italy according to the national power system and the national electric market rules. A 2013 and a 2030 scenario have been developed in order to understand effects in short and middle term. Life cycle inventory of electricity for EV has been estimated modifying available datasets according to official Italian data on power plants’ efficiency and emission rates. Finally, also for the ICEV use phase, existing dataset have been modified for fuel consumption and regulated emission according to the National Inventory Report results (ISPRA 2014) RESULTS AND DISCUSSION: In both 2013 and 2030 scenarios, the power mix that in Italy supplies energy to EV is dominated by fossil fuel power plants. Nevertheless, due to the fact that more than the 60 % of this energy is produced in efficient combined cycle gas turbine power plants, EV performs better than ICEV in almost all the impact categories considered except for human toxicity and eutrophication, the only two impact categories in which the EV car, mainly due to battery manufactory, presents more relevant potential impacts. ICEV impacts are always dominated by well to wheel phases (use phase and fuel production). EV car and battery manufacturing have higher impacts for all categories than ICEV car manufacturing. CONCLUSIONS: The study demonstrates that electricity supplied in Italy to EV today is, and will probably be in 2030, mainly produced by fossil fuel power plants. Nevertheless, the EV proves to be able to reduce, with respect to ICEV, those impacts it is supposed to reduce: air acidification, photochemical oxidant formation, and also greenhouse gases. Trade-offs are, as foreseeable, eutrophication and human toxicity due to EV car and battery manufacturing.