M.C. Veronesi

An Apgar scoring system for routine assessment of newborn puppy viability and short-term survival prognosis.

The Apgar scoring system is an easy and reliable method for evaluating both human and animal neonates. However, its use is not widespread in veterinary medicine. The current study assessed a modified Apgar scoring system for routine evaluation of newborn puppies. Heart rate, respiratory effort, reflex irritability, motility, and mucus color have been evaluated in the score. Specifically, we used 5 min after birth Apgar score to assess newborn viability and short-term survival prognosis, as well as related characteristics, in 193 puppies from 42 litters, 65 born by spontaneous delivery, 66 by assisted delivery, and 62 by cesarean section. The percentage of puppies that were dead 2h after birth was higher in the 4 to 6 Apgar score group versus that in the 7 to 10 score group (P<0.01) and in the 0 to 3 score group versus that in the 7 to 10 score group (P<0.0001). Delivery method did not affect survival. There was a marked reduction in the number of puppies searching for the mammary gland in the 0 to 3 and 4 to 6 Apgar score groups compared with that in the 7 to 10 score group (P<0.0001); there was a difference between the 0 to 3 and the 4 to 6 score groups as well (P<0.05). Suckling/swallowing reflexes were present in fewer puppies in the 0 to 3 and 4 to 6 score groups compared with that in the 7 to 10 group (P<0.0001), with no significant differences between the 0 to 3 and the 4 to 6 score groups.

Energy balance, leptin, NEFA and IGF-I plasma concentrations and resumption of post partum ovarian activity in swedish red and white breed cows

In the purpose to provide further information in respect of the relationship between metabolism and post partum (PP) ovarian activity resumption in dairy cows, the aim of the present study was to characterize the energy balance (EB) and leptin, NEFA and IGF-I plasma levels in Swedish Red and White (SRW) cows with and without ovarian activity re-initiation within 7 weeks PP. The study was conducted on 12 primiparous SRW cows fed the same diet as total mixed ration for ad libitum intake. The EB was calculated weekly from parturition until seven weeks PP. Blood samples were collected weekly from one week before until 7 weeks after calving for leptin, NEFA and IGF-I analysis. For progesterone (P4) analysis, blood samples were collected two times per week from parturition until the end of the study. P4 profile was used in addition to the clinical examination to detect cows with and without ovarian activity resumption. The clinical and ultrasonographic examination, coupled with P4 profile analysis showed the resumption of ovarian activity within 7 weeks after calving in 8 (group A) and no ovarian resumption in 4 cows (group B). No significant differences were detected in the whole period of observation in the amount of milk production between the two groups, while the mean milk protein content was significantly lower in group B at the third week PP. The calculated EB was negative in both groups in the first three weeks after calving, but more marked in group B. NEFA and Leptin plasma levels did not show significant differences between the two groups. In conclusion, the results of the present study showed that, when low milk producing primiparous cows are concerned, no significant differences in BW loss, milk yield, EB and leptin and NEFA plasma levels between the cows with and without resumption of ovarian activity within 7 weeks post partum were seen. However, significantly higher IGF-I levels in the first two weeks after calving were found in cows with post partum ovarian activity resumption, highlighting the important role of IGF-I as sensitive signal between metabolism and reproduction.

Agreement between ultrasonographic classification of the CL and plasma progesterone concentration in dairy cows

We evaluated the agreement between ultrasonographic characteristics of the corpus luteum (CL) and plasma progesterone (P4) concentration in dairy cows. In Phase I of the study, the ovaries of 8 cows were ultrasonographically examined, and P4 was analyzed daily from estrus (Day 0) to Day 4, then at Day 7 and Day 10, and again daily from Day 17 to the onset of next estrus. In Phase 2, the ovaries of 157 randomly selected Friesian cows were examined once by ultrasonography, and blood samples collected concurrently were analyzed for plasma P4. On the basis of the P4 values, the function of CLs was classified as follows: 1) non-secretory CL when plasma P4 was lower than 1 ng/mL (n=41); 2) evolving CL when plasma P4 was between 1 and 4 ng/mL (n=55); and 3) mid-cycle CL when plasma P4 was more than 4 ng/mL (n=61). On the basis of ultrasonographic examination, 3 additional groups were established (absence of CL, evolving CL, midcycle CL). Ultrasonographic characteristics and size of Day 3 to 4 CLs and their respective plasma P4 concentrations were not distinguishable from those of CLs observed 3 to 4 d before the subsequent estrus. The degree of agreement between the two classification was 72%. The data indicate that the functional classification of CLs is difficult to determine based on ultrasonography alone.

Isolation of Canine Mammary Cells With Stem Cell Properties and Tumour‐Initiating Potential

Recent data suggest that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self-renewal or by progenitor cells that have acquired the capacity for cell self-renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long-term non-adherent culture conditions. We showed that cells obtained from spheres that display self-renewing properties, have multi-lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour-initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue.

Ultrasonographic appearance of tissue is a better indicator of CL function than CL diameter measurement in dairy cows

In this study, the ovaries of 99 randomly selected Friesian cows were examined by ultrasonography measuring the diameter and evaluating the appearance of corpora lutea (CLs) in order to assess the most reliable method for their functional classification. Concurrently, blood samples were taken and analyzed for plasma progesterone (P4) concentration. On the basis of the ultrasonographic measurement of the diameter of the CL, three groups were established: (A) CL not detected (n = 30), (B) CL psi < 20 mm (n = 22), and (C) CL psi > or = 20mm (n = 47). On the basis of the ultrasonographic appearance, three different groups were established: (A) CL not detected (n = 30), (B) evolving CL (n = 25), and (C) mid-cycle CL (n = 44). On the basis of the P4 values, CLs were functionally classified in the following three groups: (A) CL not detected when plasma P4 was lower than 1 ng/ml (n = 27), (B) evolving CL when plasma P4 was between 1 and 4 ng/ml inclusive (n = 29), and (C) mid-cycle CL when plasma P4 was more than 4 ng/ml (n = 43). The degree of agreement between plasma P4 concentrations and either ultrasonographic classification (diameter or appearance) was highly significant (P < 0.001). However, the results of the present study suggest that for the evaluation of functional classification of the CL in cows ultrasonographic appearance is more reliable than the evaluation of the diameter.

Oxytocin, vasopressin, prostaglandin F2α, luteinizing hormone, testosterone, estrone sulfate, and cortisol plasma concentrations after sexual stimulation in stallions

This experiment was designed to determine the effects of sexual stimulation on plasma concentrations of oxytocin (OT), vasopressin (VP), 15-ketodihydro-PGF(2alpha) (PG-metabolite), luteinizing hormone (LH), testosterone (T), estrone sulfate (ES), and cortisol (C) in stallions. Semen samples were collected from 14 light horse stallions (Equus caballus) of proven fertility using a Missouri model artificial vagina. Blood samples were collected at 15, 12, 9, 6, and 3 min before estrous mare exposure, at erection, at ejaculation, and at 3, 6, and 9 min after ejaculation. Afterwards, blood sampling was performed every 10 min for the following 60 min. Sexual activity determined an increase in plasma concentrations of OT, VP, C, PG-metabolite, and ES and caused no changes in LH and T concentrations. The finding of a negative correlation between C and VP at erection, and between C and T before erection and at the time of erection, could be explained by a possible inhibitory role exerted by C in the mechanism of sexual arousal described for men.

Hair cortisol level as a retrospective marker of hypothalamic-pituitary-adrenal axis activity in horse foals.

Stimulation of the hypothalamic-pituitary-adrenal (HPA) axis and elevated cortisol concentrations in fetal plasma are associated with foal maturity, viability and adaptation to independent life. However, non-invasive measurement of cortisol in hair samples has not yet been validated in horses. The current study developed a radioimmunoassay to analyse cortisol in horse hair and was used to measure cortisol hair concentration at birth and at 30 and 60 days of age as a retrospective study of HPA axis activity. Cortisol was detectable in the hair of foals from birth until 2 months, but decreased with time and varied greatly between individuals. Analysis of hair cortisol could be useful for non-invasive retrospective studies of HPA axis activity in perinatal horse.

The use of oxytocin for the reduction of cow placental retention, and subsequent endometritis

Oxytocin was administered to reduce incidence of retained placenta and uterine infections that could delay subsequent conception. Three hundred and fifty multiparous Friesian cows, each with spontaneous delivery of a single calf were divided randomly into two groups. Some (n = 175) were injected with 30 IU of oxytocin immediately after delivery and again 2-4 h later, while the remainder formed an untreated control group. The placental retention 24 h after parturition was 24.6% and 10.9% in control and treated animals respectively (P < 0.01). Endometritis occurred in 51.6% of the animals following placental retention as compared to 10.4% of those with normal expulsion of the fetal membranes (P < 0.001). A comparison of reproductive indices showed a statistically significant improvement of fertility in treated cows with the average interval from calving to conception being reduced from 124.4 d to 93.7 d (P < 0.0001).

Immunohistochemical Evaluation of the Expression of Anti-Müllerian Hormone in Mature, Immature and Neoplastic Canine Sertoli Cells

In mammals, the earliest specific protein expressed by Sertoli cells (SCs) is the anti-Müllerian hormone (AMH), which induces the regression of Müllerian ducts and is produced by SCs until the functional maturation of the testes. The aim of this study was to evaluate the expression of AMH by canine SCs during testicular maturation and neoplastic transformation. Testes from two fetuses, 18 newborn puppies, five puppies aged 43-180 days and six adult dogs, and 24 canine Sertoli cell tumours (SCTs) were studied immunohistochemically for expression of AMH. Fifteen of the 24 SCTs were classified as typical, eight as lipid-rich and one was considered malignant based on evidence of lymph node metastasis. SCs from fetuses and neonatal puppies and puppies up to 45 days old expressed AMH, while SCs from older puppies and adults were negative. All SCTs expressed AMH, suggesting that AMH expression is a useful marker of immature and neoplastic canine SCs.

Efficiency of different extenders on cooled semen collected during long and short day length seasons in Martina Franca donkey.

Artificial insemination with cooled semen is routine in equids because of its good fertility rates and relatively low costs. In several donkey breeds, especially in restricted populations, the use of cooled semen could be seen as the best way of improving reproductive performance and avoiding excessive inbreeding. Furthermore, most jennies have ovulatory estrous throughout the year, and thus, cooled semen could also be used during short day length season. The aims of this study were to evaluate the effects of different extenders on sperm quality during cooling in the Martina Franca breed, and to verify the preservation of cooled semen collected during long day length (May-June) and short day length (November-December) seasons. Three ejaculates were collected at 10-day intervals from each of six jackasses during both May-June and again in November-December time periods. Each ejaculate was cooled in INRA96 or E-Z Mixin at a low cooling rate and evaluated daily over a 120-h preservation time. The results showed a significant extender influence on preservation time in both periods. Semen diluted with INRA96 maintained a progressive motility of 36% and a straightness of 89% at 120h, whereas semen extended with E-Z Mixin had a mean progressive motility of 32% and a straightness of 81% at 48h during the May-June period. Despite having the same initial characteristics, semen collected during the short day length season had a higher rate of decline in semen quality during storage at 5 degrees C with E-Z Mixin.