Neonatal-Maternal Bodyweight Ratio at Birth: An Indicator for First-Week Survival Prognosis in Canine Newborns.

Objectives. In dogs, neonatal bodyweight (NBW) varies at birth. Size-specific reference values for NBW are essential to correctly manage newborns. The present study aimed to define a new tool to evaluate NBW that could be routinely applied in canine neonatology, promptly identifying puppies requiring more care. Methods. The records of 89 litters were retrospectively evaluated. Data were grouped according to maternal bodyweight (MBW) in five categories: toy (≤5 kg), small (5.1-9.9 kg), medium (10-24.9 kg), large (25-39.9 kg), and giant (≥40 kg) dogs. At birth, the NBW of 412 puppies, alongside their litter size (LS) and gender (G), were recorded. Neonatal mortality within the first week was considered. The influence of MBW, LS, and G on NBW and the effect of NBW on the 1-week outcome were evaluated using ANOVA (p < 0.05). The ratio between NBW and MBW expressed as a percentage (N-MBW%), was assessed for each puppy. Through Receiver-Operating Characteristic (ROC) curves, N-MBW% thresholds between puppies alive or dead at one week of age were calculated in each group. Results. The LS was different among groups, except for small vs. medium dogs. In small litters, N-MBW% was lower in medium-, large- and giant-sized dogs than in toy and small dogs. In medium and large litters, N-MBW% differed among the five groups. Male and female N-MBW% differed among the five groups. Only in large and giant sizes did LS affect N-MBW%, which was lower in a large LS. The N-MBW% was higher in healthy puppies than in those who died within one week. The ROC-derived N-MBW% thresholds were as follows: 4.5% in toy-, 3.2% in small-, 1.5% in medium-, 1.2% in large-, and 1% in giant dogs. Clinical significance. The ROC-derived N-MBW% thresholds can be used to cautiously identify puppies at increased risk for one-week mortality and may be helpful in clinical practice, representing a suitable parameter to give a prognosis on 1-week survival immediately at birth.

Highlights on the Canine Prostatic Specific Esterase (CPSE): A diagnostic and screening tool in veterinary andrology.

In the last years, the need to look for an accurate and precise diagnosis of prostatic diseases in dogs has grown. Among other diagnostic tools, the seric CPSE has been studied and identified as a valid and specific biomarker for prostatic disorders, since it can result significantly more elevated in dogs affected by several prostatic abnormalities, such as benign prostatic hyperplasia, bacterial prostatitis and prostatic carcinoma. Therefore, dosing CPSE in serum represents a new diagnostic and screening tool. Dosing CPSE in everyday clinical practice has three objectives: (a) the diagnosis of benign prostatic hyperplasia; (b) the preventive screening of prostatic disorders in healthy dogs; (c) the medical follow-up in subjects with prostatic disorders during and after medical therapy. Neither circadian rhythms nor transrectal palpation performed during the andrological examination do affect CPSE. A sexual rest of at least 24 hr before dosing CPSE is recommended as it is affected by ejaculation.

Pre- and Probiotics to Increase the Immune Power of Colostrum in Dogs.

Wide differences in Ig concentration in canine colostrum have been reported. Thus, some litters can be at risk of passive immune transfer failure. Present study evaluated if supplementation with MOS, FOS, E. faecium and L. acidophilus along pregnancy increases colostrum quality. Twenty Great Dane bitches were divided into 4 groups. Control group (CG) received standard diet, only. Diet was supplemented with pre- and probiotics in other 3 study groups during: the last (1WG), last 2 (2WG), and last 4 (4WG) weeks of pregnancy, until parturition. Serum samples were collected at estrous (T0), supplementation beginning (T1), and parturition (T2). Colostrum was collected at C-section end. The IgG, IgM, and IgA were assayed on both matrices. In serum, IgG were higher at T0 than at parturition in all study groups and they significantly lowered from T0 to T1 in all groups. In colostrum, IgG and IgM were significantly higher in 4WG, while IgA already increased in 2WG group. Four-week pre- and probiotic supplementation resulted in the best immune properties of colostrum, as by the higher IgG, IgM, and IgA colostrum levels found in 4WG. Further studies would verify the exact mechanisms involved: pre-partum IgG mammary accumulation and B-cells GALT proliferation and mammary transfer. Further trials would verify whether these beneficial effects of pre- and probiotics on colostrum also lead to improved clinical conditions and immunological functions of newborns and puppies.

Pregnancy Effect on Echocardiographic Parameters in Great Dane Bitches.

Pregnancy is associated with adaptation of the left ventricular (LV) function. Due to differences between breeds in baseline echocardiographic values and specific predispositions for myocardial diseases, breed-specific echocardiographic parameters may be helpful to evaluate whether the systolic function varies during pregnancy. This study enrolled nine healthy Great Dane bitches with uncomplicated pregnancy. Echocardiographic M-mode and B-mode data were collected before ovulation and within 7 days of the predicted parturition term. Evaluated parameters were: LV dimension in diastole (LVd) and systole (LVs), end-diastolic (EDVI) and end-systolic (ESVI) volumes indexed to body surface area (BSA), end-diastolic (EDV) and end-systolic (ESV), end-point-septal-separation (EPSS), left atrium to aortic root ratio (LA/Ao), sphericity index (SI), ejection fraction (EF), fractional shortening (FS), stroke volume (SV), heart rate (HR), and cardiac output (CO). The ANOVA showed a statistical effect of the age of gestation (p < 0.01) on the increase of diastolic dimensions and functional parameters and on the decrease of systolic dimensions. The CO increase parallels the rise in SV and HR (p < 0.01). No statistical differences were observed for EPSS, LA/Ao, and SI. The changes in cardiac chambers and function are likely to reflect maternal adaptation to allow the fetal development in uncomplicated pregnancy. The present study provides specific echocardiographic values in uncomplicated pregnancy of Great Danes, showing that the systolic function is enhanced and that the increase in preload, observed during gestation, is the likely mechanism.

A Biotic Support During Pregnancy to Strengthen the Gastrointestinal Performance in Puppies.

Up to 60% of neonates can be affected by gastroenteritis due to specific pathogens or aspecific polymicrobial interactions. The present study evaluated if a dietary supplementation with MOS, FOS, E. faecium and L. acidophilus in pregnancy may reduce gastroenteritis in puppies. Fifteen Great Danes were divided in 3 groups. The control group (CG) ate a standard diet. In 2 study groups, the diet was supplemented with pre- and probiotics during the last (1WG) and the last 4 pregnancy weeks (4WG). Up to 9 weeks, puppies were checked daily to identify first- or second- presentation gastroenteritis. Data were processed by χ2 (P < 0.05). First-presentation gastroenteritis was more frequent in CG than in 1WG than in 4WG. Second-presentation gastroenteritis was more frequent in CG than in 1 and 4WG. Puppies from pre- and probiotics supplemented bitches were less prone to gastroenteritis. 1 or 4WG equally reduced second-presentation gastroenteritis in puppies, but 4WG was better than 1WG on first-presentation gastroenteritis. By entero-mammary link, supplemented bitches produced higher immune quality colostrum, thus puppies faced immunitary challenges better; moreover, maternal microbiota, positively altered by supplementation, was transferred to newborns, becoming more resistant to gastroenteritis. This information can be useful in clinical practice with the goal of preventing gastroenteritis in puppies and reducing its prevalence and severity.

Urinalysis in Great Dane Puppies from Birth to 28 Days of Age.

Urinalysis, a common test in infants, could represent a suitable non-invasive clinical tool in puppies. In dog neonates, urine is easily collected by stimulating the somato-vesccal reflex. Information on urine characteristics during the neonatal period is missing. Beside instrumental laboratory analyses, the dipstick was proven useful for rapid urinalysis to evaluate specific gravity (SG), pH, leukocytes, nitrites, glucose, proteins, ketones, urobilinogen, bilirubin, and blood. The present study aimed to describe urinalysis features by the dipstick test and refractometer along the neonatal period. Urine samples (n = 624) were collected by manual stimulation from 48 healthy Great Danes, daily from birth to seven days, then twice a week until 28 days, to assess age-related changes (ANOVA, p < 0.05) and the possible effects of gender and litter (T-test, p < 0.05). The SG and pH significantly changed during the neonatal period. Other parameters did not vary significantly in relation to age. No significant differences were observed either among litters or between genders. The present study confirmed that canine kidneys are able to concentrate urine from the second week of age when the urinary SG started to be similar to adults, while pH still increased towards the typical values of adults at 28 days. Significant glucosuria and proteinuria were never detected. Dipstick urinalysis represents a useful first-line complementary tool in newborns clinical examination, providing information about systemic homeostasis.

Ejaculation Effect on Canine Prostatic Specific Esterase Serum Concentration.

Canine prostatic diseases are usually asymptomatic in their onset and often identified in advanced stages. Canine prostatic specific esterase (CPSE) represents an early serum marker for prostatic diseases, also in asymptomatic dogs. The present study aimed to identify the effects of ejaculation on serum CPSE. Twenty adult intact male dogs were enrolled. Blood samples were collected to measure CPSE concentrations before (T0), immediately after (T1), and 24 h post (T2) ejaculation. Data were compared within and between groups by ANOVA (p < 0.05). Dogs were divided in two equal groups: A (healthy: CPSE ≤ 52.3 ng/mL at T0) and B (suspected for prostatic disorders: CPSE > 52.3 ng/mL or diagnosed with symptoms of prostatic diseases: CPSE > 90 ng/mL). CPSE was shown to be statistically higher in group B than A at any time point. In both groups, CPSE showed a significant increase at T1, and no significant differences between T0 and T2. This study demonstrates a definite effect of ejaculation on CPSE concentration. Twenty-four hours post-ejaculation, CPSE returns to basal values. Such physiological effects of ejaculation should be considered when planning analyses of CPSE concentrations, i.e., by respecting a proper sexual rest.

Effects of Alfaxalone or Propofol on Giant-Breed Dog Neonates Viability During Elective Caesarean Sections.

Attention must be paid to C-section anesthesia effects on mother and offspring. Alfaxalone induction results in improved puppy viability when compared to propofol. The present study aims to evaluate effects of alfaxalone or propofol induction for elective C-section on newborns, expressed as Apgar score and fetal fluids cortisol concentration. Anesthesia was induced with alfaxalone 3 mg/kg iv in 5 bitches (group A), and propofol 4 mg/kg iv in another 5 (group P), maintained with isoflurane. Amniotic and allantoic fluids were collected to determine cortisol concentration. Apgar score, litter size, newborn gender, birth-weight, maternal age, and parity were recorded. ANOVA, U Mann-Whitney test and ANCOVA assessed the effects of drugs on the Apgar score and fetal fluids cortisol. Thirty-six puppies were randomly selected for the study: 16 from group A and 20 from group P. Only the Apgar score significantly differed between groups. ANCOVA confirmed a significantly higher Apgar score in group A underlining the influence of fetal fluids cortisol concentrations, both resulting in covariates. Present results confirm the effect of anesthesia on the Apgar score of newborns, which is significantly higher for alfaxalone than propofol, suggesting the use of fetal fluids cortisol as a covariate. These findings could be a starting point for further investigations when less viable puppies are detected or expected, such as during an emergency C-section.

Fertility outcome after medically treated pyometra in dogs.

Cystic endometrial hyperplasia-pyometra complex (CEH/P) is a challenge in canine reproduction. Present study aimed to assess fertility after medical treatment. One-hundred-seventy-four bitches affected by CEH/P received aglepristone on days 1, 2, 8, then every 7 days until blood progesterone < 1.2 ng/mL; cloprostenol was administered on days 3 to 5. Records were grouped according to bodyweight (BW): small (< 10 kg, n = 33), medium (10 ≥ BW < 25 kg, n = 44), large (25 ≥ BW < 40 kg, n = 52), and giant bitches (BW ≥ 40 kg, n = 45). Age; success rate; aglepristone treatments number; relapse, pregnancy rates; diagnosis-relapse, -first, -last litter intervals; litters number after treatment, and LS were analyzed by ANOVA. Overall age was 5.14 ± 1.75 years, without difference among groups. Treatment was 100% successful, without difference in treatments number (4.75 ± 1.18), relapse (15/174, 8.62%) and pregnancy (129/140 litters, 92.14%) rates, intervals diagnosis-relapse (409.63 ± 254.9 days) or -last litter (418.62 ± 129.03 days). The interval diagnosis-first litter was significantly shorter (163.52 ± 51.47 days) and longer (225.17 ± 90.97 days) in small and giant bitches, respectively. Overall, 1.47 ± 0.65 litters were born after treatment. Expected LS was achieved in each group, as shown by ΔLS (actual-expected LS by breed, overall -0.40 ± 1.62) without differences among groups. Concluding, CEH/P affects younger dogs than previously described. Relapses were rarer than previously reported. Medical treatment with aglepristone+cloprostenol is effective and safe, preserving subsequent fertility, as demonstrated by negligible changes in LS.

Assessment of the optimal age for a preventive ultrasonographic screening of the uterine health in bitches.

Studies about prevalence of uterine pathologies in bitches are scarce. Although correlation between age and uterine disorders was documented, the most suitable age for a preventive sonographic screening has not been proposed yet. Present study aimed to estimate the eligible age for an ultrasonographic screening of uterine abnormalities in dogs. Data regarding ultrasound examination and clinical records of non-pregnant intact females were retrospectively analysed. The age of each bitch was expressed as age ratio (actual/maximum age expected for the respective breed). The cut-off age ratio was determined by a ROC curve for overall uterine abnormalities. Frequencies of different abnormalities below and over the cut-off derived from the ROC curve were calculated and statistically analysed by chi-Square and OR. Prevalence of three categories of ultrasonographic findings was as follows: cystic endometrial hyperplasia (CEH) 18%; uterine collections (UC) 10.5%; masses (M) 1.3%. By the cut-off age ratio (0.325), derived by ROC curve (AUC = 0.91; SP 84.23%; SE 79.2%; PPV 83.4%; NPV 80%), 228 cases were divided into two subgroups: bitches over (exposed group: n.83) and below cut-off (control group: n.145). All abnormalities resulted more frequent in exposed group: OR was 24.96 (p < 0.0001: 71.1% over vs. 9% below cut-off) for overall abnormalities; 13.68 (p < 0.0001: 40.9% vs. 4.8%) for CEH; 6.13 (p < 0.002: 21.7% vs. 4.1%) for UC; 12.65 (p = 0.09: 3.6% vs. 0%) for M. Cystic endometrial hyperplasia represents the most common finding in adult bitches, followed by UC. A preventive sonographic screening for uterine abnormalities should start from 33% of expected longevity to preventively select animals requiring further evaluations.

The Effect of Dietary Supplementation of Vitamin E, Selenium, Zinc, Folic Acid, and N-3 Polyunsaturated Fatty Acids on Sperm Motility and Membrane Properties in Dogs.

Sub-fertility represents a common challenge in canine reproduction. Different protocols, supplementing daily given quantities of micronutrients, were investigated to improve poor sperm concentration and/or function, which represent breeding major constraining factors in the canine species. Little information is available for dogs concerning the effect of a daily supplementation with a complex of vitamin E, zinc, selenium, folic acid, and n-3 polyunsaturated fatty acids (PUFA) on semen quality. Thus, the present study investigated this effect on semen motility and sperm membrane properties. Serial semen analyses from fourteen healthy normospermic dogs, fed with the same commercial diet, were performed on Days 0 (T0), 30 (T30), 60 (T60), and 90 (T90). Seven dogs were randomly included in the treatment (T) group, receiving a supplementation of vitamin E, zinc, selenium, folic acid, n-3 PUFA; and seven other subjects composed the control (C) group. Total Sperm Count (TSC), Computer-Assisted Sperm Analysis (CASA) indexes, mortality, and functional membrane integrity were assessed. The ANOVA compared results between groups and sampling times (p < 0.05). From T60, the supplementation significantly improved TSC, progressive motility, functional membrane integrity, and decreased mortality. Present results lead us to consider ameliorative effects of a two-month healthy diet supplementation on canine spermatozoa. The positive effects of the described balanced integration of micronutrients on sperm motility and prevention of oxidative stress should be considered, especially when decreased seminal parameters may result from inadequate intake, reduced absorption, increased losses or demand, or to attenuate the impact of age.

Effect of delivery management on first-week neonatal outcome: How to improve it in Great Danes.

A different incidence of neonatal complications has been reported in several species according to the duration of the various labour stages and the type of delivery. Trauma associated with dystocia may result in the release of neonatal catecholamines and cortisol that negatively affect neonatal metabolism, thus survival. Aim of the present study was to assess how the first-week neonatal outcome is affected by a different management of parturition. Clinical records of 46 Great Dane litters from 8 different kennels were evaluated retrospectively. Data were grouped in 3 categories: vaginal delivery (VD), emergency C-section (ES), planned C-section (PS). The risk of stillbirth, early neonatal mortality and perinatal mortality was calculated at puppy and at litter level in each group. Data were statistically compared by the ANOVA and Chi-Square test (p < 0.05). Seventy-eight puppies were naturally delivered (VD), 120 experienced dystocia at birth that required emergency C-section (ES) and 105 came from planned C-section (PS). No statistical difference in neonatal bodyweight and litter size was observed among groups. In PS, both stillbirth and neonatal mortality resulted significantly lower than in VD and ES, resulting in the lowest observed perinatal mortality. The number of litters with stillbirth was significantly higher in ES than VD and PS. The number of litters with neonatal mortality resulted significantly lower in PS than VD and ES. The overall perinatal mortality in VD resulted statistically influenced from both stillbirth and neonatal mortality, while it resulted strongly affected by the high stillbirth in ES. No difference was observed for PS perinatal mortality. Present results, reporting a high incidence of dystocia as well as high perinatal and neonatal mortality, lead us to evaluate the opportunity to plan an elective C-section in Great Dane bitches, in order to minimize perinatal mortality.

Advances in Prostatic Diagnostics in Dogs: The Role of Canine Prostatic Specific Esterase in the Early Diagnosis of Prostatic Disorders.

In last years, following the increased canine life expectancy and the rising attention pet-owners devote to their animals, several authors have carried on investigations concerning new techniques to early identify canine prostatic disorders that might affect the dog's quality of life. Prostatic disorders often have an asymptomatic onset and their early diagnosis is difficult: hence, they are usually identified at an advanced stage, only. Traditionally, the diagnosis of prostatic disorders is based on noninvasive tools, such as transrectal and abdominal palpation, seminal or prostatic fluid evaluation, and urinalysis and imaging. On the other hand, a definite diagnosis of prostatic abnormalities could be achieved through prostatic parenchyma Fine Needle Aspiration (FNA) or biopsy. However, these investigations are performed rarely because of their invasiveness. Thus, several authors investigated canine serum biomarkers in order to achieve an earlier diagnostic timing and to apply therapeutic strategies for better outcomes. The Canine Prostatic Specific Esterase (CPSE) has been identified as a suitable biomarker to be included in a prostate health screening program, following the model of prostate-specific antigen (PSA) in human medicine. A higher CPSE in dogs suffering from several prostatic diseases, such as benign prostatic hyperplasia, bacterial prostatitis, or prostatic carcinoma, was reported in literature. Thanks to the potential usefulness in clinical practice, further studies should investigate the potential role of CPSE in monitoring the medical treatment success in the male reproductive system. Moreover, the spreading availability of serum biomarkers, easily carried out on blood samples in clinical practice, could assure a more accurate evaluation of the actual prevalence of prostatic disorders. The CPSE is actually recognized as a promising diagnostic tool for the detection of prostatic disorders in a "prostate health screening program," in order to properly select those patients requiring further more accurate and expensive diagnostic investigations.

Ejaculation effect on blood testosterone and prostatic pulsed-wave Doppler ultrasound in dogs.

Local vessels ultrasonography evaluates prostatic physio-pathologic states. Testosterone promotes tissue and vascular growth. Knowing variables on prostatic vasculature is crucial to correctly apply Pulsed-Wave exam. The study aims to assess how ejaculation and blood testosterone affect Pulsed-Wave indexes. Serial blood testosterone dosages and Pulsed-Wave exams were performed in 20 dogs, immediately before (T0) and after (T1) ejaculation and 6 hr later (T2). Arteria prostatica cranialis, Arteriola capsularis, Arteriola trabecularis and Arteriola parenchimalis were evaluated and mean Pulsatility and Resistivity Index, Systolic-Peak, End-Diastolic and Mean Velocity calculated. Data were grouped by time and vessel (ANOVA, p ≤ 0.05). At T1, Resistivity Index significantly lowered in A. prostatica cranialis, A. trabecularis and A. parenchimalis but grew in A. capsularis; Pulsatility Index had the same pattern, but not significant in A. parenchimalis; Systolic Peak Velocity, End-Diastolic Velocity, Mean Velocity significantly rose in A. capsularis and A. trabecularis. No indexes differed at T0 and T2. Testosterone did not differ at T0 (10.93 ± 7.05 ng/ml), T1 (12.71 ± 7.29) and T2 (10.54 ± 6.63). Results stated the risen prostatic vascular flow postejaculation, affecting Pulsed-Wave. Due to semi-rigid capsule, impairing vasodilation of other vessels, only A. capsularis indexes increased. Intimal cushions of A. prostatica cranialis kept velocities fixed; A. capsularis and A. trabecularis lack of intimal cushions, thus velocities grew. In A. parenchimalis, precapillary sphincters opening allows increased flow redistribution in vasodilated parenchymal bed, keeping velocities fixed. As testosterone, not affected by ejaculation, did not peak, vascular changes are not due to testosterone itself. These physiological effects of ejaculation suggest proper sexual rest before Pulsed-Wave exam planned to explore suspected prostatic neovascularization.

Colour Flow Mapping examination: An useful screening test for the early diagnosis of ductus venosus patency in canine newborns.

Ductus venosus (DV) closure plays a key role in hepatic circulation adaptation to postnatal metabolic function, and DV patency might develop a congenital portosystemic shunt (CPSS). The noninvasive Color Flow Mapping (CFM) examination, a validated method to diagnose CPSS in adult dogs, is routinely performed to assess DV closure after birth in humans. This study aimed to describe the feasibility of the ultrasonographic evaluation of the DV after birth and to determine its closure time in healthy Great Dane neonates. Patency of DV in serial Color Flow Mapping (CFM) examinations and bodyweight (BW) were recorded on Days 0-3-6-9 in 24 neonates that were classified as having patent (PDV) or closed ductusvenosus (CDV) basing on CFM signal presence/absence. Since the 3rd day, DV diameter was recorded. Data were analysed by ANOVA (p < 0.05). All dogs resulted healthy 1 year later. The number of PDV and CDV puppies at birth was not different on Day 3 (24 and 0 vs. 22 and 2, PDV and CDV, respectively), whereas it resulted different on Days 6 (24 and 0 vs. 14 and 10) and 9 (24 and 0 vs. 0 and 24); on Day 3, it was different compared to Days 6 and 9; on Day 6, it was different from Day 9. Reduction of DV diameter resulted positively related to neonatal BW growth. The CFM evaluation of DV closure after birth in Great Dane puppies represents a feasible technique. Present results suggest the time of functional closure in normal neonates within 9 days after birth. Thus, CFM examination, as an early screening test for DV patency evaluation, performed 10 days after birth, may identify suspicious dogs at risk that would require further investigations. Further studies are needed to deepen the role of a delayed closure in low bodyweight and preterm puppies.