Materials and Methods

The present study was conducted in the whole Italian territory. Certified canine breeders by ENCI (Italian National Kennel Club) were recruited at dog shows and through social media breeder's groups and invited by email to complete an anonymous online questionnaire on breeding management practices. The inclusion criteria for this study included dog breeders currently operating in the Italian territory who voluntarily agreed to participate, acknowledged the confidentiality and anonymity of their responses and fully completed the online questionnaire about their breeding management practices. Data collection was performed from October 2023 to March 2024. The questionnaire was administered with the aim of covering the entire Italian territory. The questionnaire consisted of 41 questions requiring short or numerical answers about where the breeding facility was located, how many breeds were kept in the facility for breeding purposes, and how the heats, pregnancies and newborn puppies were managed (how heats were monitored and with which techniques, if artificial insemination was performed, if pregnancy diagnosis and follow‐up was done, and how newborn puppies were managed until weaning). Once the questionnaires were completed, data were analysed, and incomplete questionnaires were removed from the study. As questionnaires were anonymous, breeders could not be contacted in the case of incomplete or incomprehensible information provided. Incomplete questionnaires, those with contradictory or unintelligible information, were excluded from the final analysis.

Using the sample size calculator (Qualtrics), the number of respondents was established to ensure that the sample is adequate and representative of Italian dog breeders. As no animals were manipulated for data collection, no ethical committee approval was needed.

Statistical Analysis

All data collected through the questionnaire were entered into Microsoft Excel and subsequently analysed using RStudio software (version 2025.09.0). Descriptive statistics were calculated for all categorical and continuous variables. The categorical variables were summarized as absolute and relative frequencies (percentages), whereas continuous variables were reported as means ± standard deviations (SD). The distribution of each continuous variable was assessed for normality using Shapiro–Wilk test, and the homogeneity of variances was evaluated using either Bartlett's test (for normally distributed data) or Levene's test (for non‐normally distributed data). To investigate regional differences, breeders were grouped into three geographical macro areas: North (Valle d'Aosta, Piemonte, Liguria, Lombardia, Trentino‐Alto Adige, Veneto, Friuli‐Venezia Giulia and Emilia‐Romagna), Centre (Toscana, Umbria, Marche, Lazio, Abruzzo and Molise) and South (Campania, Puglia, Basilicata, Calabria, Sicilia and Sardegna). For categorical variables, contingency tables were constructed, and the chi‐square test of independence (χ 2) was applied to assess associations between each variable and the geographical macro area. When the expected frequency in any cell was below 5, Fisher's exact test was considered for validation. Continuous variables were compared among macro areas using the Kruskal–Wallis test. When significant differences were detected, appropriate post hoc analyses were conducted using the Dunn test for pairwise comparisons with Bonferroni or Holm correction when appropriate. A p value lower than 0.05 was considered statistically significant.