Discussion

To the author's knowledge, this is the first descriptive study that documents canine breeders’ approach to reproductive performance in a Southern European country.

This nationwide survey of 251 Italian dog breeders offers a comprehensive snapshot of current reproductive and early‐life management practices across Northern, Central and Southern Italy. Overall, breeders reported mostly consistent approaches to oestrus monitoring, pregnancy diagnosis and puppy care, with regional differences mainly in how intensely and when they conduct monitoring rather than in their core practices.

Breeder and Breed Distribution

Although there are no official data on the number of breeders in Italy in 2024, an estimate based on ENCI data suggests that there are approximately 1700 registered kennels. Dividing Italy into three macro areas, showing the number of kennels and the most representative breeds in the study, made it easier to interpret the data, providing a satisfactory overall view (Figures 1 and 2).

Heat Detection Techniques

For both veterinarians and breeders, the identification of the optimal day of mating is crucial to achieve a higher pregnancy rate. This study highlights that all three macro areas show a substantially overlapping trend in preferences in the use of the different techniques employed to identify the optimal moment for mating of the female. The canine oestrous period is longer than in other species, and clinical symptoms during the oestrus cycle stages are non‐specific. The whole heat (pro‐oestrus and oestrus) can last 3–4 weeks, whereas the best time for mating is only 3–4 days (Concannon and Rendano 1983). The female can accept the male even in pro‐oestrus, thus causing the fertilization of the gametes to fail. In this study, data show that the use of the male to monitor oestrus is falling out of use throughout Italy. In fact, all the breeders answered use progesterone to monitor the bitch. Determining the LH peak requires several blood samples each day until its detection, whereas P4 increases progressively during heat (Hase et al. 2000). Together with the longer oestrus cycle, another issue in canine reproduction is that oocytes are immature just after ovulation and need around 48–72 h for complete maturation and for being fertilizable (Chastant‐Maillard et al. 2011; Reynaud et al. 2005). Ovulation detection is crucial for good reproductive management. Therefore, P4 currently seems to be the most accurate and easily detectable hormone to estimate ovulation. However, there is still no single P4 concentration value that definitively determines ovulation in bitches, as P4 concentrations vary significantly depending on the measurement method used (Gloria et al. 2018; Nöthling and De Cramer 2018; Zuercher et al. 2021). The near‐universal P4 monitoring in our cohort study (96.4%) underscores the role of hormonal assays in breeding management even if around 70% of breeders do not know the detection method used. Although this may not be critical since it is the veterinarian's responsibility to understand the analytical system to correctly identify ovulation, it is interesting to note that the average P4 concentration when the breeder decided to mate the bitch or perform artificial insemination was around 12.8 ± 4.00 ng/mL. Even if most of breeders did not know what machine were their vets using for P4 measurement, these results are in accordance of the usage of a chemiluminescence or fluorescence assay, as ovulation tends to be detected between 4 and 10 ng/mL (Brugger et al. 2011; Gloria et al. 2018; Milani et al. 2022; Rota et al. 2016; Tal et al. 2020). The high prevalence detected on P4 assays reflects the actual clinical recommendations that emphasize P4 as the key marker for ovulation detection for insemination optimization. On the other side, comparative evaluations of P4 machines (RIA vs. chemiluminescence/FEIA/CLIA) showed that modern CLIA/chemiluminescent assays correlate well with good‐standard methods, support high reliability when used appropriately (Milani et al. 2022) but may require assay‐specific thresholds for clinical decision‐making (Nöthling and De Cramer 2018, 2019; Tal et al. 2020). This variability underscores why progesterone values alone should not be interpreted without knowledge of the specific assay used, a point of practical relevance for both clinicians and breeders. On the basis of questionnaire responses, vaginal cytology was performed and evaluated by the veterinarian in approximately by half of respondents with modest regional variations starting between the second and third day of vulvar discharge with an average of two vaginal smears (Figures 3 and 4). More than 50% of breeders using vaginal cytology as a test for oestrus staging consider this technique valuable during oestrus monitoring (Figure 4). Vaginal cytology remains an inexpensive and fast in‐clinic technique for oestrus monitoring, often used together with P4 measurements. Despite breeders’ generally positive perception of this technique, vaginal cytology interpretation is performed by the veterinarian and requires specific training; the literature reports poor inter‐observer agreement on vaginal cytology cell‐type classification (Moxon et al. 2010; Reckers et al. 2022) suggesting that breeders’ favourable view of this tool may not fully reflect its technical reliability, and that it should always be complemented by hormonal data. In fact, the proportion of breeders using vaginal cytology in our study is consistent with reports that many practitioners and breeders combine cytology and P4 for optimal timing (Barstow et al. 2018; Fáy et al. 2003; Roos‐Pichenot and Zakošek Pipan 2025). Vaginoscopy was the least used technique, as almost 90% of breeders considered it useless. As physiological background, both techniques, vaginal cytology and vaginoscopy, might be useful to estimate the oestrus cycle phase: The rise in oestradiol during pro‐oestrus increases vaginal mucosa swelling and crenulation, and a progressive increase in keratinized cells occurs as oestrus approaches. Once the oestrus phase is reached, more than 90% of epithelial cells are superficial keratinized cells, most with pyknotic nuclei (Bouchard et al. 1991). Nevertheless, both vaginoscopy and vaginal cytology, whether used alone or combined, are unable to determine the exact moment of ovulation. The comparison of mating practices between regions (Figure 5) does not show marked differences among the macro areas considered. It must be considered that up to 80% of breeding failures in the canine population are attributed to incorrect timing of mating or insemination (Roos‐Pichenot and Zakošek Pipan 2025). Breeders’ educational programmes are often lacking across Europe, and no harmonized regulatory approach to breeder training as yet been established.

Mating and Type of Semen Preferences

Over all the responses received, natural mating remained the predominant method across all the macro areas studied (>55%). VI was used by only a third of the respondents, whereas TCI was uncommon and surgical insemination was absent in the South of Italy and rare in central and northern Italy (Figure 6). These distributions mirror wider clinical practice where natural mating and VI with fresh semen remain common, whereas TCI and surgical intrauterine techniques are commonly used primarily for chilled or frozen semen (Romagnoli and Lopate 2014; Mason 2018).

The strong preference for fresh semen across regions (>90%) is consistent with established evidence showing higher conception rates with fresh semen compared with frozen or chilled semen when insemination is optimally timed (Figure 7). The low use of frozen semen reported by breeders in our sample may reflect their concern about the additional costs and logistical demands associated with TCI, which is generally required to achieve adequate conception rates with frozen‐thawed semen (Cicirelli et al. 2021; Hollinshead and Hanlon 2017; Linde‐Forsberg et al. 1999; Silva et al. 1995; Thomassen et al. 2006). Although well‐timed TCI performed by trained clinicians can yield satisfactory pregnancy rates even with frozen semen, the higher cost and need for specialized expertise likely discourage breeders from adopting this approach on a routine basis. These technical and logistical demands might likely explain the sustained preference for semen among breeders (Joonè 2024; Linde‐Forsberg et al. 1999). Nevertheless, several reports indicate higher pregnancy rates with well‐timed TCI using frozen semen in specific settings and breeds, highlighting that TCI provides an effective, non‐surgical intrauterine option when performed by trained clinicians (Pretzer et al. 2006). Nonetheless, logistical cost and training barriers limit TCI adoption at the breeder level. The limited demand for frozen and chilled semen suggests that the use of imported genetic material remains relatively low in Italy. This fact is of particular interest as, over time, this restricted genetic exchange may contribute to reduced genetic diversity within breeds, increasing the risk of inbreeding and limiting the introduction of desirable reproductive or health‐related traits.

Puppy Management Practices

Pregnancy diagnosis was performed on average at the age of 26.55 ± 4.84 days from expected ovulation (Figure 4) with ultrasonography widely adopted (>90%), consistent with its recognized role in early, non‐invasive pregnancy confirmation (Lopate 2018). Most of the breeders (87%) demanded the x‐ray puppy count, with Southern breeders performing x‐rays slightly earlier on average, reflecting different scheduling practices or a tendency for early imaging to plan whelping logistics, but both ranges fall within the commonly accepted window for reliable foetal skeletal visualization (Concannon and Rendano 1983). The timing differences, while statistically significant, were modest, and their clinical relevance should be interpreted with caution. In fact, earlier radiographs may yield less accurate foetal counts if the gestational age is inadequate. Interestingly, the questionnaire responses showed that breeders in Central and Southern Italy reported performing fewer diagnostic tests overall than breeders in Northern Italy. This difference may reflect regional variations in access to specialized reproductive services (Santos et al. 2021).

Routine weighing of puppies to monitor growth was common, but frequency varied regionally: Northern breeders weighed puppies more often compared to Central and Southern (Figures 8 and 9). Daily increase in weight is significant for puppy survival, especially in low‐weight puppies at birth, and to detect early failure to thrive, guide supplementary feeding and improve survival (Mugnier et al. 2023). Thus, regular weighing is clue to monitor puppies’ growth and detect early problems. Daily weighing should be performed during the first 2 weeks of puppies’ life and every 2–3 days until approximately the month of age (Alves 2020; Kirk 2001). Overall, puppies with a birth weight under 25% of the average litter birth weight have a 12 times higher mortality rate during the first week of life (Mila et al. 2015; Mugnier et al. 2019). In addition, significant weight variations during the first 48 h of life (a loss of ≥4% of birth weight) have been reported to increase puppies’ mortality on the first 3 weeks of age (Mila et al. 2015). Weaning of puppies was performed on average at 28 days of life, and the use of milk replacer as a supplement was relatively uncommon and showed limited regional variation. This suggests that most breeders rely on maternal lactation and reserve milk replacer for indicated cases, aligning with neonatal care recommendations that caution against unnecessary supplementation.