Frameshift Variant in AMPD2 in Cirneco dell'Etna Dogs with Retinopathy and Tremors.

While the manifestations of many inherited retinal disorders are limited to loss of vision, others are part of a syndrome that affects multiple tissues, particularly the nervous system. Most syndromic retinal disorders are thought to be recessively inherited. Two dogs out of a litter of Cirneco dell' Etna dogs, both males, showed signs of retinal degeneration, along with tremors and signs described as either atypical seizures or paroxysmal dyskinesias, while the other two male littermates were normal. We named this oculo-neurological syndrome CONS (Cirneco oculo-neurological syndrome), and undertook homozygosity mapping and whole-genome sequencing to determine its potential genetic etiology. Notably, we detected a 1-bp deletion in chromosome 6 that was predicted to cause a frameshift and premature stop codon within the canine AMPD2 gene, which encodes adenosine monophosphate deaminase, an enzyme that converts adenosine 5'-monophosphate (AMP) to inosine 5'-monophosphate (IMP). Genotyping of the available Cirneco population suggested perfect segregation between cases and controls for the variant. Moreover, this variant was absent in canine genomic databases comprised of thousands of unaffected dogs. The AMPD2 genetic variant we identified in dogs presents with retinal manifestations, adding to the spectrum of neurological manifestations associated with AMPD2 variants in humans.

Identification of a common haplotype in carriers of rob(1;29) in 32 Italian cattle breeds.

Robertsonian translocation 1;29 (rob(1;29)), a widespread chromosomal anomaly affecting cattle fertility, appears to have originated from a common ancestor. This study utilizes routine SNP data to investigate the chromosomal region associated with rob(1;29) and confirm the presence of a shared haplotype among carriers in diverse Italian breeds. Three datasets were employed: Dataset 1 included 151 subjects from 5 beef cattle breeds genotyped with the GGP Bovine 33 k SNP chip; Dataset 2 encompassed 800 subjects from 32 Italian breeds genotyped with the Illumina 50 k SNP chip, sourced from the BOVITA dataset; Dataset 3 combined Dataset 2 with 21 karyologically tested subjects from breeds with a high carrier frequency, genotyped using the Affymetrix 65 K SNP chip. FST analysis pinpointed a distinctive genomic region on the first six Mb of BTA29, the centromeric region involved in the translocation. Haplotype comparisons within this non-recombining region revealed a common haplotype shared among all carriers, supporting the theory of a common ancestor. Principal component and haplotype analysis allowed clear differentiation of rob(1;29) homozygous and heterozygous carriers. Expanding to Dataset 2 revealed rob(1;29) carriers in unexpected breeds, all sharing the same ancestral haplotype. Notably, previously untested breeds, including Cinisara, exhibited a high carrier prevalence (nearly 50%), confirmed by karyological analysis. This study validates the presence of a shared haplotype among all identified rob(1;29) carriers, reinforcing the common ancestor theory as the origin of this translocation's spread throughout the cattle population. Furthermore, it underscores the potential of SNP data analysis as a rapid, accurate, and cost-effective tool for broad rob(1;29) screening, given the translocation's consistent nature across all analyzed breeds.

Phenotypic and Genomic Characterization of the Comune di Sicilia Goat: Towards the Conservation of an Endangered Local Breed.

The Comune di Sicilia, a local goat breed from Sicily (Italy), is currently undergoing recognition as a distinct breed. This study aims to characterize the population both phenotypically and genomically to advance its recognition process. A total of 78 subjects from two locations were enrolled, and their phenotypic data, including qualitative traits and morphometric measurements of adult animals, were recorded and statistically analyzed. The goats were genotyped using the Illumina 50 k Goat SNPchip, comparing them with 473 goats from 15 Italian breeds. Population structure, phylogenetic relationships, admixture, and genomic inbreeding were analyzed. Additionally, subjects with different morphological traits were compared using FST and runs of homozygosity, leading to the identification of potential candidate genes associated with anotia and wattle presence in goats. The Comune di Sicilia breed exhibited distinctive genomic and phenotypic features, setting it apart from other breeds in the same region. However, moderate variability, possibly influenced by selection practices, was also observed. To ensure the breed's preservation and prevent excessive inbreeding, a comprehensive approach considering both morphology and genomic background is recommended. This study contributes valuable insights into the genetic peculiarities of the Comune di Sicilia goat, supporting its recognition as a unique and valuable breed.

The Shepherd and the Hunter: A Genomic Comparison of Italian Dog Breeds.

Shepherd and hunting dogs have undergone divergent selection for specific tasks, resulting in distinct phenotypic and behavioural differences. Italy is home to numerous recognized and unrecognized breeds of both types, providing an opportunity to compare them genomically. In this study, we analysed SNP data obtained from the CanineHD BeadChip, encompassing 116 hunting dogs (representing 6 breeds) and 158 shepherd dogs (representing 9 breeds). We explored the population structure, genomic background, and phylogenetic relationships among the breeds. To compare the two groups, we employed three complementary methods for selection signature detection: FST, XP-EHH, and ROH. Our results reveal a clear differentiation between shepherd and hunting dogs as well as between gun dogs vs. hounds and guardian vs. herding shepherd dogs. The genomic regions distinguishing these groups harbour several genes associated with domestication and behavioural traits, including gregariousness (WBSRC17) and aggressiveness (CDH12 and HTT). Additionally, genes related to morphology, such as size and coat colour (ASIP and TYRP1) and texture (RSPO2), were identified. This comparative genomic analysis sheds light on the genetic underpinnings of the phenotypic and behavioural variations observed in Italian hunting and shepherd dogs.

Effects of Dietary Enrichment with Olive Cake on the Thyroid and Adrenocortical Responses in Growing Beef Calves.

Agro-industrial by-products incorporated into livestock feed formulations can positively impact feed costs and promote a circular bio-economy. Italy produces significant amounts of olive cake (OC), a by-product of olive oil extraction, with the potential for incorporation into bovine diets. However, information on its effects on endocrine responses in growing beef calves is lacking. Forty-eight Limousines randomly allocated to dietary treatment (control or 10%-OC or 15%-OC inclusion) were segregated according to sex and body weight. Serum concentrations of TSH, thyroid hormones, and cortisol were measured on day 0, day 56, and at the end of the trial on day 147. Circulating TSH, total (T3, T4) and free (fT3, fT4) iodothyronines, and cortisol concentrations were all within the normal physiological ranges, with no significant effect imparted by diet. However, the diet × time interaction was significant for T3. The cortisol, T3, T4, and fT4 registered on day 147 were higher than those of day 56, and cortisol was higher in heifers than bulls. Final body weight was positively correlated with TSH and T3 and negatively with cortisol concentration. These findings suggest that the inclusion of OC at levels up to 15% in growing/finishing beef diets had no adverse effects on the calves' thyroid and cortical status.

Genomic Analysis of the Endangered Fonni's Dog Breed: A Comparison of Genomic and Phenotypic Evaluation Scores.

The Fonni's dog is an ancient Sardinian breed for livestock and property guarding. In recent years, the number of new registrations to the breeding book has slumped and, thus, this breed risks being lost forever. This work refocuses attention to the Fonni's dog, analysing its genomic makeup and comparing different phenotypical and genetic evaluation scores. Thirty Fonni's dogs were ranked by their general accordance to the breed typicality (typicality score) and to the provisional standard by official judges (judges' score). They were genotyped with a 230K SNP BeadChip and compared with 379 dogs of 24 breeds. Genomically, the Fonni's dogs placed themselves near shepherd dogs and showed a unique genetic signature, which was used to create the genomic score. This score better correlated with typicality (ρ = 0.69, p < 0.0001) than the judges' score (ρ = 0.63, p = 0.0004), which showed little variability among the included dogs. Hair texture or colour were significantly associated in the three scores. The Fonni's dog is confirmed as a well-distinguished breed, despite being selected mainly for its work abilities. The evaluation criteria used during dog expositions can be improved to increase their variability and include elements typical of the breed. The recovery of the Fonni's dog would be possible only with a shared vision between the Italian kennel club and breeders, and the support of regional programs.

Evaluation of Chemical Contaminants in Conventional and Unconventional Ragusana Provola Cheese.

Organic contaminants belonging to various classes (plasticizers, bisphenols, pesticides, PCBs, and PAHs,) were analyzed in samples of provola cheese produced from Friesian dairy cows fed with a conventional diet (group CTR), and an unconventional diet (group BIO) enriched with olive cake (OC). The results show that for most determined contaminants, the differences between the two diets were very slight, indicating that the contamination does not depend on the olive cake integrated in the unconventional diet. The results also indicate that the minimal contamination could result from environmental contamination or the production process. It can be concluded that unconventional provola is as safe for the consumer as conventional provola.

Using Pedigree and Genomic Data toward Better Management of Inbreeding in Italian Dairy Sheep and Goat Breeds.

The inbreeding coefficient is an important parameter for livestock management. Small ruminant breeders and associations mainly rely on pedigree information, but genomic tools are gaining relevance, overcoming possible pedigree inconsistencies. This study investigates the relationship between pedigree-based and genomic inbreeding in two goat and four sheep dairy breeds. Pedigree and genomic data (medium-density SNPchip) were obtained for 3107 goats and 2511 sheep. We estimated pedigree depth (number of fully traced generations, FullGen) and inbreeding (FPED), as well as two genomic inbreeding indexes, using runs of the homozygosity (FROH) and genomic relationship matrix (FGRM). The correlation between the inbreeding coefficients was assessed. A linear regression model (LRM) was fitted for estimating FPED from FROH. After quality control on genomic data, we retained 5085 animals. Mean inbreeding values were low, with higher FROH than FPED and FGRM. Breed differences can partially depend on different managements. The correlation between FPED and FROH was the highest and directly related to pedigree depth. The best LRM was chosen for FullGen ≥4 and ≥6 for goats and sheep, respectively; after excluding animals with extreme residuals, a new refined regression equation was calculated. Since massive genotyping is not affordable to small ruminant breeders, it is important to understand the distinction and relationship between differently calculated inbreeding coefficients, also in view of the introduction of genomic enhanced breeding values. Our study highlights the importance of accurate pedigree information and, especially if not obtainable, of calculating genomic-based inbreeding coefficients. A better estimation of animals' relatedness contributes to improve animal breeding and conservation.

Changes of acute-phase proteins, glucose, and lipid metabolism during pregnancy in lactating dairy cows.

The study aimed to evaluate the effects of different stages of lactation (0 to >  300 d) and pregnancy (0 to >  180 d) on serum amyloid A (SAA), C-reactive protein (CRP), glucose, total cholesterol (TCho), and triglyceride (TG) concentrations in dairy cows of different breeds. Thus, 40 healthy multiparous cows (10 Holstein, 10 Simmental, 10 Brown, and 10 Modicana) were randomly selected, and blood samples were collected once every 60 d for 1 year. Overall, SAA and CRP serum concentrations progressively increased and became more variable along the lactation, peaking at >  240-300 d, and then decreased in the last period ( >  300 d). Along pregnancy, SAA and CRP initially increased, with the highest concentrations at >  60-120 d, and then decreased until the last phase of pregnancy ( >  180 d). However, lactation and gestation phases did not significantly affect SAA and CRP when all the cows were analyzed together. A significant and positive correlation was observed between SAA and CRP both along lactation ( r = 0.89 ; p < 0.0001 ) and pregnancy ( r = 0.91 ; p < 0.0001 ). Breeds only showed differences in CRP levels along gestation ( p = 0.0102 ), due to a peak registered at 0-60 d in Holstein cows. In pregnant cows, glucose was positively correlated with SAA ( r = 0.43 ; p = 0.0017 ) and CRP ( r = 0.42 ; p = 0.0019 ). Hence, these significant and positive relationships reflect the physiological adaptations of the dairy cows along both gestational and lactational dynamics, suggesting that these proteins may also be involved in non-pathological processes. In this perspective, this study established that the obtained response markedly varies among healthy individuals along lactation and gestation and thus that the physiological range of acute-phase proteins (APPs) is wide; this makes it difficult to use these proteins as a marker of different physiological reproductive and productive periods.

Adaptive Responses of Thyroid Hormones, Insulin, and Glucose during Pregnancy and Lactation in Dairy Cows.

The study examined the effects of different stages of pregnancy and lactation on thyroid, insulin, and glucose responses in dairy cows. In the present study, 30 dairy cows (10 Holstein, 10 Simmental, and 10 Brown) at 30 ± 20 d of lactation were randomly selected and blood samples were collected once every 60 d for one year to measure circulating thyroid stimulating hormone (TSH), total and free triiodothyronines (T3, fT3) and thyroxines (T4, fT4), insulin, and glucose. Pregnant cows showed higher T4 (p = 0.010) and insulin (p = 0.046) concentrations at >180 d than at >60−120 d of pregnancy and in nonpregnant cows. Along the lactation phase, circulating T4 concentrations showed a biphasic trend, decreasing from 0−60 d to >60−120 d phase, which showed the lowest values, and then increasing until the end of lactation (>300 d) (p = 0.016). Glucose showed the highest concentrations at the start of lactation (0−120 days) and the lowest values at the end (p = 0.008). The monitoring of thyroid hormones, insulin, and glucose changes represents an important tool to evaluate the anabolic and/or catabolic adaptation in response to functional periods in dairy cows, which can potentially predispose the cows to an excessive negative energy balance and related metabolic issues.

Selection Signatures in Italian Livestock Guardian and Herding Shepherd Dogs.

Livestock guardian (LGD) and herding shepherd (HSD) dogs have distinct morphological and behavioural characteristics, long selected by farmers and breeders, to accomplish different tasks. This study aimed to find the genomic regions that best differentiate and characterise Italian LGD and HSD. Genomic data of 158 dogs of four LGD and five HSD breeds, obtained with the 170K canine SNPchip, were collected. The two groups were compared using FST and XP-EHH analyses, identifying regions containing 29 genes. Moreover, 16 islands of runs of homozygosity were found in LGD, and 15 in HSD; 4 of them were partially shared. Among the genes found that better differentiated HSD and LGD, several were associated with dog domestication and behavioural aspects; particularly, MSRB3 and LLPH were linked to herding behaviour in previous studies. Others, DYSK, MAP2K5, and RYR, were related to body size and muscle development. Prick ears prevailed in sampled HSD, and drop ears in LGD; this explains the identification of WIF1 and MSRB3 genes. Unexpectedly, a number of genes were also associated with eye development and functionality. These results shed further light on the differences that human selection introduced in dogs aimed at different duties, even in a limited geographic area such as Italy.

Steroid hormonal endpoints in goats carrying single or twin fetuses reared in semi-extensive systems.

The study provides baseline data regarding 17- ß -estradiol (E 2 ), progesterone (P 4 ), and cortisol profile of 30 Nicastrese goats during different physiological periods. Animals were evaluated monthly from the pre-mating period (non-pregnant), during pregnancy, and from 30 to 105 d of lactation. The effects of single or twin births and the kid's sex were also considered. Serum E 2 , P 4 , and cortisol concentrations were measured using immunoenzymatic assay kits. The highest concentrations of E 2 and P 4 ( P < 0.0001 ) were found during pregnancy and their lowest values ( P < 0.0001 ) in the non-pregnant period. E 2 was negatively correlated with P 4 ( r = - 0.41 ; P < 0.01 ) during lactation. The mothers with twin kids showed the highest concentration of P 4 ( P < 0.04 ) at >  95-115 d of gestation and the lowest of E 2 ( P < 0.04 ) at >  50-70 d of lactation. Pregnant goats carrying male kid(s) presented the highest E 2 concentrations ( P < 0.02 ) at >  130-150 d of gestation. Different physiological conditions induced a temporal relationship with the endocrine profile in Nicastrese goats. Understanding the effects of single or twin fetuses on the gestation and lactation will also be helpful to improve the managemental approach for the health of mothers and their kids.

Runs of homozygosity in the Italian goat breeds: impact of management practices in low-input systems.

BACKGROUND: Climate and farming systems, several of which are considered as low-input agricultural systems, vary between goat populations from Northern and Southern Italy and have led to different management practices. These processes have impacted genome shaping in terms of inbreeding and regions under selection and resulted in differences between the northern and southern populations. Both inbreeding and signatures of selection can be pinpointed by the analysis of runs of homozygosity (ROH), which provides useful information to assist the management of this species in different rural areas. RESULTS: We analyzed the ROH distribution and inbreeding (FROH) in 902 goats from the Italian Goat Consortium2 dataset. We evaluated the differences in individual ROH number and length between goat breeds from Northern (NRD) and Central-southern (CSD) Italy. Then, we identified the signatures of selection that differentiate these two groups using three methods: ROH, ΔROH, and averaged FST. ROH analyses showed that some Italian goat breeds have a lower inbreeding coefficient, which is attributable to their management and history. ROH are longer in breeds that are undergoing non-optimal management or with small population size. In several small breeds, the ROH length classes are balanced, reflecting more accurate mating planning. The differences in climate and management between the NRD and CSD groups have resulted in different ROH lengths and numbers: the NRD populations bred in isolated valleys present more and shorter ROH segments, while the CSD populations have fewer and longer ROH, which is likely due to the fact that they have undergone more admixture events during the horizontal transhumance practice followed by a more recent standardization. We identified four genes within signatures of selection on chromosome 11 related to fertility in the NRD group, and 23 genes on chromosomes 5 and 6 related to growth in the CSD group. Finally, we identified 17 genes on chromosome 12 related to environmental adaptation and body size with high homozygosity in both groups. CONCLUSIONS: These results show how different management practices have impacted the level of genomic inbreeding in two Italian goat groups and could be useful to assist management in a low-input system while safeguarding the diversity of small populations.

Influence of Morphometry on Echocardiographic Measurements in Cavalier King Charles Spaniels: An Inverse Probability Weighting Analysis.

The development and progression of myxomatous mitral valve disease (MMVD) in Cavalier King Charles Spaniels (CKCS) are difficult to predict. Thus, the identification of dogs with a morphotype associated with more severe mitral disease at a young age is desirable. The aims of this study were to: (1) describe the physical, morphometric, and echocardiographic features of class B1 MMVD-affected Cavalier King Charles Spaniels (CKCS) according to the American College of Veterinary Internal Medicine (ACVIM) guidelines; (2) evaluate the influence of morphometric physical measurements on murmur intensity, mitral valve prolapse (MVP), regurgitant jet size, and indexed mitral valve and annulus measurements. Fifty-two MMVD-affected CKCS were included in the ACVIM class B1. This is a prospective clinical cross-sectional study. Morphometric measurements, which included the body, thorax, and head sizes of each dog, were investigated to establish the association with heart murmur intensity, valvular and annular echocardiographic measurements, MVP, and regurgitant jet size, using inverse probability weighting (IPW) analyses to adjust for confounding. The IPW analyses showed that when the head length and nose length decreased, dogs had a more severe regurgitant jet size. Furthermore, subjects with a more pronounced head stop angle had thicker anterior mitral valve leaflets. A brachycephalic morphotype, as seen in dogs similar to the King Charles Spaniel breed in terms of cephalic morphology, is associated with a more severe regurgitant jet size and with valvular characteristics that are related to the most severe forms of MMVD.

Genetic trend of the junctional epidermolysis bullosa in the German shorthaired pointer in Italy.

BACKGROUND: Epidermolysis bullosa (EB) is a hereditary heterogeneous group of mechanobullous disorders caused by mutations in several structural skin proteins observed in both humans and animals. In this work, we report the incidence and the genetic trend of the junctional epidermolysis bullosa (JEB), a major type of EB, in the Italian German Shorthaired Pointer (GSPs) population in a 10 years span. METHODS: In this study, we monitored the genetic trend of JEB in the Italian population of the GSPs from 2009 to 2018 in 750 animals. The studied mutation was the insertion (4818+207 ins 6.5 kb) of repetitive satellite DNA within intron 35 of the LAMA3 gene. RESULTS: Allele frequencies showed a reduction of the mutated (C) allele during the years, with the only exception of 2017, when 13 dogs were diagnosed as carrier for the genetic pathology. A regression logistic analysis was performed, including sex, coat colour and their interaction. Our results showed that there was a statistically significant association with coat colour. CONCLUSIONS: The simplicity and the low cost of the analysis for the detection of this pathology suggests that a deeper identification of carrier dogs will allow better breeding strategies and management, leading to a rapid JEB eradication.

Do You Think I Am Living Well? A Four-Season Hair Cortisol Analysis on Leisure Horses in Different Housing and Management Conditions.

The satisfaction of leisure horses' behavioral needs has begun to be considered a priority, linked to the awareness that horses kept in single boxes may be deprived of social contact and the possibility to perform natural behaviors. Several factors may influence horses' quality of life also in the paddock, and there are very few data on the effects of those variables on leisure horses' chronic stress, measurable in terms of activation of the hypothalamic-pituitary-adrenocortical axis. Therefore, managerial choices faced by owners and stables managers are only based on experience, common sense, and anecdotal beliefs. This study assessed and compared the chronic stress levels in leisure horses hosted in structures in the same geographic and climatic area with different daily routines to verify which management strategy could be the one that better contributes to achieving the welfare of horses. Forty-seven horses were divided into three groups homogeneous in terms of sex and age: Mixed management group (n = 12), Paddock group (n = 19), and Natural management group (n = 16). The hair cortisol concentration, a reliable marker of long-term stress, was analyzed in all the horses the same day at four time points of the year. In addition to management strategies, the influences of other variables (sex, age, coat color, and season) were evaluated. Independently from the management strategies, significantly higher hair cortisol values were detected in the autumn and summer, as well as in individuals older than 15 years. No significant differences were highlighted between the sexes or the coat colors. The comparison of the different management strategies showed that, in the summer, autumn, and winter, the hair cortisol levels were significantly lower in the Mixed management group horses than the Paddock group, highlighting that those subjects had better homeostasis. The Natural management group horses' hair cortisol levels were intermediate between the other two groups of horses in all the seasons. Spending the night in the stables would seem to positively impact the well-being of the horses. These findings, if confirmed by further studies, may be helpful in enhancing horse welfare and assisting in managerial choice decision-making.

Thyroid and Lipidic Profiles in Nicastrese Goats (Capra hircus) during Pregnancy and Postpartum Period.

This study aimed to determine the thyroid and lipid profiles in 30 Nicastrese goats, along different physiological periods: before mating (nonpregnant goats), during the whole pregnancy (pregnant goats), and during postpartum and early lactation (milking goats). Blood samples were collected monthly from March 2020 to January 2021. Serum thyroid-stimulating hormone (TSH), total and free triiodothyronine (T3, fT3), and thyroxine (T4, fT4) concentrations were measured using immunoenzymatic assay kits and serum lipid panels (triglyceride (TG) and total cholesterol (tCho)) by enzymatic colorimetric method; very-low-density lipoprotein cholesterol (VLDL Cho) was calculated. Pregnant and milking goats showed the lower T3 (p < 0.0002) and T4 (p < 0.0005) concentrations, with lower BCS (p < 0.001) only in pregnant ones. Milking goats showed tCho (p < 0.006) concentrations lower than nonpregnant ones, and TG and VLDL Cho (p < 0.001) lower than both pregnant and nonpregnant goats. T4:T3 and T3:fT3 were significantly and positively correlated in both pregnancy and lactation. Under similar environmental, nutrition, and management conditions, different physiological phases play a significant role in the thyroid and lipid profiles in Nicastrese goats. These endocrine and metabolic resources could contribute to the knowledge useful for the salvage of this endangered, local, native breed.

The climatic and genetic heritage of Italian goat breeds with genomic SNP data.

Local adaptation of animals to the environment can abruptly become a burden when faced with rapid climatic changes such as those foreseen for the Italian peninsula over the next 70 years. Our study investigates the genetic structure of the Italian goat populations and links it with the environment and how genetics might evolve over the next 50 years. We used one of the largest national datasets including > 1000 goats from 33 populations across the Italian peninsula collected by the Italian Goat Consortium and genotyped with over 50 k markers. Our results showed that Italian goats can be discriminated in three groups reflective of the Italian geography and its geo-political situation preceding the country unification around two centuries ago. We leveraged the remarkable genetic and geographical diversity of the Italian goat populations and performed landscape genomics analysis to disentangle the relationship between genotype and environment, finding 64 SNPs intercepting genomic regions linked to growth, circadian rhythm, fertility, and inflammatory response. Lastly, we calculated the hypothetical future genotypic frequencies of the most relevant SNPs identified through landscape genomics to evaluate their long-term effect on the genetic structure of the Italian goat populations. Our results provide an insight into the past and the future of the Italian local goat populations, helping the institutions in defining new conservation strategy plans that could preserve their diversity and their link to local realities challenged by climate change.

A Genomic Study of Myxomatous Mitral Valve Disease in Cavalier King Charles Spaniels.

Cavalier King Charles spaniels (CKCSs) show the earliest onset and the highest incidence of myxomatous mitral valve disease (MMVD). Previous studies have suggested a polygenic inheritance of the disease in this breed and revealed an association with regions on canine chromosomes 13 and 14. Following clinical and echocardiographic examinations, 33 not-directly-related CKCSs were selected and classified as cases (n = 16) if MMVD was present before 5 years of age or as controls (n = 17) if no or very mild MMVD was present after 5 years of age. DNA was extracted from whole blood and genotyped with a Canine 230K SNP BeadChip instrument. Cases and controls were compared with three complementary genomic analyses (Wright's fixation index-FST, cross-population extended haplotype homozygosity-XP-EHH, and runs of homozygosity-ROH) to identify differences in terms of heterozygosity and regions of homozygosity. The top 1% single-nucleotide polymorphisms (SNPs) were selected and mapped, and the genes were thoroughly investigated. Ten consensus genes were found localized on chromosomes 3-11-14-19, partially confirming previous studies. The HEPACAM2, CDK6, and FAH genes, related to the transforming growth factor ß (TGF-ß) pathway and heart development, also emerged in the ROH analysis. In conclusion, this work expands the knowledge of the genetic basis of MMVD by identifying genes involved in the early onset of MMVD in CKCSs.