Timing and Patient Position During Cuff Blood Pressure Measurement Affect Myocardial Work Parameters Measured by Echocardiography.

BACKGROUND: Although cuff blood pressure measurement is a critical parameter to calculate myocardial work noninvasively, there is no recommendation about when and how to measure it. Accordingly, we sought to evaluate the effects of the timing during the echo study and the patient's position on the scanning bed during the cuff blood pressure measurement on myocardial work parameter calculations. METHODS: One hundred one consecutive patients (44 women, 66 ± 14 years) undergoing clinically indicated echocardiography were prospectively enrolled. During the echocardiographic study, we measured the cuff blood pressure 4 times, using a fully automatic digital blood pressure monitor applied to the right and left arm in the same position throughout the study: BP1, before the start of the echo study, with the patient lying in the supine position; BP2, after positioning the patients on their left side to start the echo study; BP3, at the time of the acquisition of the 3 apical views (4- and 2-chamber and long-axis) used to measured left ventricular global longitudinal strain; and BP4, at the end of the echo study with the patient again in the supine position. RESULTS: Systolic blood pressureat BP1 was 147 ± 21 mm Hg. Between BP1 and BP2, it dropped by 17 ± 9 mm Hg (P < .05). Systolic blood pressure at BP3 was significantly lower than BP2 (130 ± 20 mm Hg vs 122 ± 18 mm Hg, P < .05), and at BP4 was significantly lower than at BP1 (-9 ± 13 mm Hg, P < .05). The average global longitudinal strain was -16% ± 3%. Accordingly, the global work index was 1,929 ± 441 mm Hg% at BP1, dropped to 1,717 ± 421 at BP2, decreased to 1,602 ± 351 mm Hg% at BP3, and increased to 1,815 ± 386 mm Hg% at BP4 (P < .001). CONCLUSIONS: The timing during the echocardiography study and the patient's position on the scanning bed are critical determinants of the measured cuff systolic blood pressure and the resulting values of myocardial work parameters.

GnRH analogue triptorelin acetate induces ovulation in jennies during oestrus.

Most of the northern hemisphere donkey breeds are faced with the risk of extinction, thus donkey reproduction is considered an emerging branch of theriogenology, and the management of artificial insemination and induction of ovulation is a crucial point in setting up preservation protocols. For four consecutive cycles, six jennies' ovarian activity was routinely monitored; an ultrasound examination was performed daily from the evidence of a follicle diameter ≥27 mm until ovulation. Upon reaching a follicular diameter ≥32 ± 2 mm (Hour 0), oestrous jennies were treated alternatively with 0.1 mg triptorelin acetate, sc, (TRIP), 0.4 mg/sc of buserelin acetate (BUS) or saline, sc, (CTRL) and examined ultrasonographically at Hours 14, 24, 38, 42, 48, 62 and every 24 h until ovulation. Induction of ovulation was considered successful if ovulation occurred from 24 to 48 h after treatment. 11/12 cycles resulted in ovulation for TRIP and 12/12 for BUS and CTRL groups, respectively. Mean ± SD ovulation time after treatment was 37.3 ± 8.3, 47.1 ± 21.0 and 66.8 ± 25.9 h for BUS, TRIP and CTRL, respectively (p = .0032). Ovulation rates between h24 and h48 were 10/12 (83.3%) for both TRIP/BUS and 2/12 (16.7%) for CTRL, respectively (p = .003). Buserelin and triptorelin-treated jennies had a 25 times higher probability to ovulate between Hours 24 and 48 than controls (p = .003), while there were no jenny and cycle effects on the ovulatory rate. The results of this study show how triptorelin successfully induced ovulation in jennies, like other GnRH analogues previously evaluated.

Endometrial expression of antimicrobial peptides as markers of subclinical endometritis in mares.

BACKGROUND: Endometritis is a major cause of subfertility in mares. Multiparous old mares are more susceptible to developing endometritis given that ageing is associated with an altered immune response and with inadequate physiological uterine clearance after breeding, which can lead to degenerative changes in the endometrium. Molecules such as antimicrobial peptides (AMPs) have been proposed as endometritis markers in the equine species. STUDY DESIGN: Cross-sectional. OBJECTIVES: To investigate the endometrial expression of defensin-beta 4B (DEFB4B), lysozyme (LYZ) and secretory leukocyte peptidase inhibitor (SLPI) genes in mares either affected or not by subclinical endometritis, due to the role of these AMPs in the immune response to bacteria and inflammatory reactions. METHODS: Endometrial biopsy for histopathological and gene expression examinations was performed on 26 mares. The inclusion criteria for the normal mare group (NM, N = 7) were 2-4 years of age, maiden status, no clinical signs of endometritis and a uterine biopsy score of I, while for mares affected by subclinical endometritis (EM, N = 19) the inclusion criteria were 10-22 years of age, barren status for 1-3 years, no clinical signs of endometritis and a uterine biopsy score between IIA and III. RESULTS: A significantly higher expression of LYZ (NM: 0.76 [1.84-0.37] vs. EM: 2.78 [5.53-1.44], p = 0.0255) and DEFB4B (NM: 0.06 [0.11-0.01] vs. EM: 0.15 [0.99-0.08], p = 0.0457) genes was found in endometritis mares versus normal mares. Statistically significant moderate positive correlations were found between the level of expression of LYZ gene and both the age (r = 0.4071, p = 0.039) and the biopsy grade (r = 0.4831, p = 0.0124) of the mares. MAIN LIMITATIONS: The study investigated a limited number of genes and mares, and the presence/location of the proteins coded by these genes was not confirmed within the endometrium by IHC. CONCLUSIONS: If the results of this study are confirmed, LYZ and DEFB4B genes can be used as markers to identify mares that are affected by subclinical endometritis.

Peak Atrial Longitudinal Strain and Risk Stratification in Moderate and Severe Aortic Stenosis.

AIMS: We sought to investigate the association with outcome of left atrial strain in a large cohort of patients with at least moderate aortic stenosis (AS). METHODS AND RESULTS: we analyzed 467 patients (mean age 80.6 ± 8.2 years; 51% men) with at least moderate AS, and sinus rhythm. The primary study endpoint was the composite of all-cause mortality and hospitalizations for heart failure. After a median follow-up of 19.2 (IQR 12.5-24.4) months, 96 events occurred. Using the ROC curve analysis, the cut-off value of peak atrial longitudinal strain (PALS) more strongly associated with outcome was < 16% [AUC 0.70 (95% CI: 0.63-0.78), p<0.001]. The Kaplan Meier curves demonstrated a higher rate of events for patients with PALS<16% (log-rank p<0.001). On multivariable analysis, PALS [aHR 0.95 (95% CI 0.91 - 0.99), p=0.017] and age were the only variables independently associated with the combined endpoint. PALS provided incremental prognostic value over left ventricular (LV) global longitudinal strain, LV ejection fraction, and right ventricular function. Subgroup analysis revealed that impaired PALS was independently associated with outcome also in the subgroups of paucisymptomatic patients [aHR 0.98 (95% CI 0.97 - 0.98), p=0.048], moderate AS [aHR 0.92, (95% CI 0.86 - 0.98), p=0.016], and low-flow AS [aHR 0.90, (95% CI 0.83 - 0.98), p=0.020]. CONCLUSION: In our patients with at least moderate AS, PALS was independently associated with outcome. In asymptomatic patients, PALS could be a potential marker of subclinical damage, leading to better risk stratification, and, potentially, to earlier treatment.

Collection of epididymal semen in the tomcat (Felix catus) by stereomicroscope-aided retrograde flushing (SARF) improves sample quality.

In small and laboratory animals epididymal spermatozoa can be collected only by the mincing technique however, blood and cell debris contamination may adversely affect the quality of the samples. This study aimed at performing the retrograde flushing technique, for the first time, in the tomcat. In the first experiment attempts to insert a blunted needle into tomcats' ductus deferentes were made and the cannulation time and the success rate were recorded. In the second experiment, insertion of the needle into the ductus was carried out under a stereomicroscope for one testis, while the other was submitted to mincing for sperm recovery and the samples obtained from paired testes were compared. In the first study, only twenty-eight percent of the cannulation attempts were successful whereas in the second experiment, 91% of the deferentes were successfully cannulated: 80% of the ducts were cannulated in less than three minutes. The stereomicroscope-aided retrograde flushing samples showed significantly higher sperm viability, motility and velocities, as compared to those resulted from mincing. The domestic cat serves as a model for more than 36 species of wild felids; the developed stereomicroscope-aided retrograde flushing technique has the potential to become the gold standard method for harvesting spermatozoa of endangered species that have a small ductus deferens.

Embryonic resorption rates at canine pregnancy diagnoses: A retrospective evaluation.

Pregnancy diagnosis in canines is generally performed during embryonic phase, between 19 and 35 days of gestation. At this stage embryonic resorptions can be observed, which, according to the literature, affects 11-26% of conceptuses and 5-43% of pregnancies. Resorption has been hypothesized as a physiological event in uterine overcrowding, however other factors may be involved, such as infectious or non-infectious diseases. This study aimed to retrospectively evaluate the incidence of embryo resorption at ultrasonographic pregnancy diagnosis in different dog breeds, and to identify the main factors determining the occurrence of the resorption sites. 95 pregnancy diagnoses were performed 21-30 days post-ovulation by ultrasound examination on 74 different animals. Breed, weight, and age of the bitches were recorded, and the reproductive anamnesis was collected from their medical records. The overall pregnancy rate was 91.6%. In 48.3% of pregnancies (42/87), at least one resorption site was visible, and embryonic resorption rate was 14.2% (61 resorption sites/431 total structures). Binary logistic regression showed a significant effect of age (P < 0.001), but not the size of the litter (P = 0.357), nor the size of the mother (P = 0.281) or any previous reproductive problems (P = 0.077). Age was significantly higher in pregnancies with resorptions than in normal ones (60.88 ± 18.24 and 40.27 ± 15.74 months, respectively, P < 0.001). The embryonic resorption rate was in line with previous findings, while the incidence of affected pregnancies was higher. Although resorptions may occur physiologically in pregnancies with large litters, a relationship between embryo resorption and litter size was not identified in our sample group, while aging increased the resorption rates. This, together with the occurrence of repeated embryonic resorptions in some bitches included in the study, suggests how resorptions could also be the result of pathological events. The underlying mechanisms and other factors that may be involved need further clarification.

Interspecific and Intraspecific Artificial Insemination in Domestic Equids.

Horses and donkeys differ phenotypically and karyotypically, although they can interbreed freely. Eight Standardbred mares and nine Amiata donkey jennies were included in the study. Semen was collected from two horses and two donkey stallions of proven fertility. A first pregnancy diagnosis was performed on day 10 after ovulation and repeated every day until embryo detection or until day 16. Irrespectively of the sire species, pregnancy rates in horse females (20/30, 66.7%) were significantly higher than in donkey females (19/70, 27.1%) (p < 0.05), while horse and donkey males did not affect pregnancy rates. Comparing overall intraspecific and interspecific AI, pregnancy rates were 25/37 (67.6%) and 14/63 (22.2%), respectively (p = 0.0001). The lowest pregnancy rate was obtained when inseminating jennies with horse stallion semen (8/49, 16.3%). No statistical differences were found when comparing embryo diameters, day at first pregnancy diagnosis, or in vitro embryo morphological quality among groups. In this study, much poorer results were obtained with jennies than with mares. Interspecific AI resulted in lower pregnancy rates than intraspecific Al, and AI to produce hinny hybrids resulted in the lowest pregnancy rate. Further studies are required to better understand the mechanism involved in such different outcomes in relation to intra- and interspecific breeding in domestic equids.

The Kisspeptin analogue C6 induces ovulation in jennies.

Kisspeptins (KPs) are the most potent stimulating neurotransmitters of GnRH release, and consequently KP administration triggers LH and/or FSH release. In small ruminants, KP or its analogs induced an LH surge followed by ovulation in both cyclic and acyclic animals, while in the mare KP only increased LH plasma levels but failed to induce ovulation. This study in jennies compares the endocrinological effects, ovulatory and pregnancy rates of the KP analog C6 and the GnRH analog buserelin acetate. The ovarian activity of nine Amiata jennies was monitored daily by transrectal ultrasound for three complete estrous cycles. Jennies in estrus were assigned, to one of three treatment groups: 50 nmol of the KP analog C6 (injected twice, 24 h apart, C6 group); 0.4 mg buserelin acetate (injected once, Bu group); and 2 mL of saline (injected once, CTRL group). Blood samples were collected at Day-1 (-24 h) Day0 (h0, before treatment), h2, h4, h6, h8, h10, h24 (before second treatment with C6), h26, h28, h30, h32, h34, h48 and every 24 h until ovulation. Jennies were inseminated once at h24 with fresh extended semen from a donkey stallion. Pregnancy diagnoses were performed 14 days after ovulation. On days 5, 10, and 14 after ovulation, for every CL the cross-sectional area (CSA) and the vascularized area (VA) were recorded by color doppler ultrasound and measured. Significantly higher plasma LH levels were found after induction between the Bu and CTRL groups at h6 and h8 (P < 0.05), while tendentially higher differences were found between the Bu/C6 groups and CTRL at h10. Five/9, 4/9, and 2/9 jennies ovulated between 24 and 48 h after induction from the Bu, C6, and CTRL groups respectively, (P > 0.05). Correlations between corpora lutea CSA and VA with serum progesterone concentration were r = 0.31, P = 0.01, r = 0.38, P = 0.01, respectively. Pregnancy rates after artificial insemination did not differ among groups (CTRL: 6/9, 66.7%; C6: 7/9, 77.8%; Bu: 6/9, 66.7%; P > 0.05). Ovulation rates after C6 treatment were comparable to that of Bu, although not different from the CTRL. Pregnancy rates were comparable to the literature in terms of fresh extended donkey semen in every group. This study suggests that stimulation of the Kp system in jennies, in contrast to findings observed in mares, induces ovulation. Further studies using higher doses and/or more animals are needed to better characterize the efficacy of C6 in jennies.

Deep-horn Artificial Insemination With Frozen Thawed Semen After re-extension in Autologous Seminal Plasma May Improve Pregnancy Rates in Jennies.

In the literature, the very low pregnancy rates after artificial insemination (AI) with frozen semen in donkeys were improved in one study after re-extension of the frozen-thawed semen in autologous seminal plasma. The aims of our study were (1) to describe in vitro post-thaw parameters of donkey jackass semen after re-extension in seminal plasma (SP) or in INRA96 and (2) to compare pregnancy rates in jennies bred with frozen-thawed semen using two different AI protocols. Semen collected from two Amiata donkey stallions, known to be fertile, was frozen in INRA96 supplemented with 2% egg yolk, and 2.2% glycerol. From the same jacks, seminal plasma was filtered, and stored at -20°C. According to the in vitro analysis, the post-thaw re-extension in SP some kinematic parameters decreased more over time. Two different AI protocols were tested: inseminating deeply into the uterine horn (DHAI) with or without thawed semen re-extension in SP. Higher pregnancy rates were obtained with re-extension in SP than when AIs were performed without SP. Further studies are needed to confirm the advantage of re-extension in autologous SP of donkey frozen-thawed semen before AI in jennies.

Variables Affecting Veterinary Students' Ability to Accurately Interpret Ovulation in Live Mare Palpation.

In a veterinary medicine curriculum, students' hands-on practice is essential but is still considered one of the major deficiencies in veterinary schools in Europe. After theoretical and basic practical training, students, under the control of experienced veterinarians (supervisors), monitored the reproductive cycle of embryo recipients by transrectal palpation and ultrasound. To evaluate the skills of students, the question "Has she ovulated?" was posed when a dominant follicle ≥ 35 mm was recorded in the previous day's examination and a score of 1 or 0 was assigned in the case of a correct or incorrect answer (test palpation), respectively. Study 1 involved the retrospective evaluation of 3,509 test palpation records of 43 students (31 females, 12 males) and showed a statistically significant positive correlation between the number of test palpations performed and the proportion of correct answers. There was a statistically significant effect of the number of test palpations performed by each student, their gender, and the season on the correct answers. When performing > 50 test palpations, a statistical difference between gender was observed (p < .05). Study 2 involved the prospective evaluation of 687 records on 52 standardbred or thoroughbred recipient mares collected from nine right-handed female students. The different mares, breed, occurrence of ovulation on the left or right ovary, and the presence of one or more large follicle(s) per ovary had no effect on the correct answers (p > .05). Individual students' performances were statistically different (p < .05), ranging from 60% to 92%.

hCG is more effective than the GnRH agonist buserelin for inducing the first ovulation of the breeding season in mares.

BACKGROUND: Human Chorionic Gonadotropin (hCG) and Gonadotropin Releasing Hormone agonists (GnRHa) are routinely used to induce ovulation in mares. However, GnRHa efficacy in transitional mares has been suggested to be low. OBJECTIVES: The aims of this study were as follows: (a) to compare the efficacy of hCG and GnRHa in inducing the first ovulation of the breeding season and (b) to evaluate the correlation between ovulatory response, uterine oedema and teasing score at the time of treatment during the early or late transitional phase. STUDY DESIGN: Randomised controlled superiority trial. METHODS: Mares in winter anoestrus were treated with sulpiride when at least two follicles reached a diameter of 25 mm. The day after the follicle reached 35 mm in diameter, mares in oestrus were treated with GnRHa buserelin (N = 29) or hCG (N = 33) and checked daily for ovulation. RESULTS: More mares (30/33, 90.1%) ovulated when the first ovulation after winter anoestrus was induced with hCG, than with GnRHa, (11/29, 38.0%) (P = .0001). Ovulation rate was lower in mares that did not show uterine oedema and full acceptance of the teaser stallion for at least three days before the treatment (32/41, 78% vs 9/21, 42.9%) P = .01. MAIN LIMITATIONS: Plasma LH and oestrogen concentrations were not performed. CONCLUSIONS: These results demonstrate that hCG was more effective than GnRHa for inducing ovulation in the first cycle after winter anoestrus. Uterine oedema and behavioural signs of oestrus, for at least three days before the treatment, were predictors for a positive response to ovulation induction.

Maternal and Neonatal Evaluation of Derived Reactive Oxygen Metabolites and Biological Antioxidant Potential in Donkey Mares and Foals.

Our aim was to measure the concentrations of derived reactive oxygen metabolite (d-ROMs) and biological antioxidant potential (BAP) of donkey mares and foals at delivery and to verify the protective role of the placenta against fetal oxidative stress. A total of 15 Amiata jennies with a physiological gestation length and delivery were included together with 17 foals (two twin foalings). After delivery, maternal and foal venous blood samples were collected along with blood from the artery. Circulating lactate and plasma d-ROMs and BAP were evaluated. The Wilcoxon test for paired data was applied to verify differences in d-ROMs and BAP values, while the Spearman test was used to evaluate correlations. A significantly higher d-ROMs concentration was found in jennies compared to their foals, and to the umbilical artery blood. The BAP was higher in jennies than in their foals, but no differences were observed in the umbilical artery blood. No difference was found between foals and their umbilical cord. Blood lactate was higher in foals than in their dams. Positive correlations were found between mares and umbilical cord for BAP and d-ROMs, and between mares and foals and umbilical cord for BAP. In conclusion, the placenta may be a protective factor for the fetus. As with equine foals, the antioxidant system of donkey foals does not seems to be effective at birth.

Prevalence and prognosis of pericardial effusion in patients affected by pectus excavatum: A case-control study.

BACKGROUND: The presence of pectus excavatum(PEX) has been occasionally associated with pericardial effusion. Aim of the present study was to compare incidence and prognosis of pericardial effusion in a group of unselected patients with PEX vs a control group. METHODS: From a prospective registry of consecutive patients who underwent chest CT for cardiovascular disease, subjects with a radiological diagnosis of PEX were retrospectively identified (cases); from the same registry patients (controls) without rib cage abnormalities were randomly selected, until a 1:2 ratio was reached. The presence of pericardial effusion at CT was quantified. Follow-up was obtained for a composite end-point: cardiac tamponade, need for pericardiocentesis, need for cardiac surgery for relapsing pericardial effusion. RESULTS: A total of 43 patients with PEX (20 females) and a control group of 86 cases (31 females) without rib cage abnormalities were identified. Pericardial effusion evaluated at CT was significatively more prevalent in patients with PEX vs control group, 37.2% vs 13.9% (p < 0.001), respectively; four patients with PEX (9.3%) had at least moderate pericardial effusion vs no subjects among the controls (p = 0.004). PEX diagnosis was significantly associated to pericardial effusion at multi-variate analysis (OR95%CI 10.91[3.47-34.29], p < 0.001). At a mean follow-up of 6.5 ± 3.4 years no pericardial events were recorded. CONCLUSION: Our findings support the higher prevalence of pericardial effusion in patients with PEX when compared to a control group. The absence of adverse pericardial events at follow-up suggest the good prognosis of these effusions, that in the appropriate clinical setting might not be considered "idiopathic".

Gross and histological findings in the canine placenta and amnion at term: What's normal, abnormal or pathological?

The canine placenta is an underexamined organ. Placental abnormalities can affect foetus development and may be responsible for a low weight of the infant at birth; however, knowledge on their clinical significance in the canine species is limited. We aimed to describe macroscopic and microscopic findings in the canine placenta and amnion at term in clinically uncomplicated pregnancies and to evaluate their relationship with birth weight of healthy puppies. During natural delivery or C-section, the birth weight of 82 puppies was recorded, 72 placentas and 66 amnions were recovered. The foetal and maternal surfaces of the placental girdle, marginal haematoma and amnion were evaluated. Each gross finding was recorded, morphometrically assessed and sampled for histological diagnosis. Furthermore, specimens of placenta and amnion were collected from representative areas and microscopic deviations from normal structure were evaluated in haematoxylin and eosin sections. Gross examination revealed 'abnormalities' in the 75.4% of the collected placentas. Necrosis was the gross change most commonly observed in the placental girdle (72.5%). Congestion (17.4%) and clotted blood/fibrinoid material (2.9%) were also observed. No gross changes of either the marginal haematoma or the amnion were recorded. Histologically, placental girdle showed necrosis (62.3%), mineralization (52.2%), congestion (36.2%) and neutrophilic infiltration (27.5%). Marginal haematoma exhibited mineralization (11.6%) and neutrophils (29%), while necrotic foci were rarely observed (4.3%). In the amnion, the most frequent alteration observed was hypertrophy of the epithelium (35.9%) followed by oedema (31.2%), mineralized foci (28.1%), fibrosis (23.4%), congestion (15.6%) and more rarely neutrophils (12.5%). Puppies' birth weight was not statistically affected by either gross or histological abnormalities. Our study revealed that macroscopic and microscopic 'abnormalities' of the placenta and amnion may be common in uncomplicated pregnancies at term; however, no implications on puppies' birth weight were observed. Deviations from 'normal' morphology of canine foetal adnexa warrant further investigation to assess their clinical implications if present.

Hastening Time to Ejaculation in Donkey Jacks Treated with the PGF2α Analog, Cloprostenol Sodium.

Due to the long courtship needed to attain excitation and erection, donkey semen collection can take up to 90 min. ProstaglandinF2α (PGF2α) has been reported to hasten the onset of erection and ejaculation in domesticated mammals, presumably by inducing smooth muscle contractions in the internal genitalia. However, while it has been anecdotally used in donkeys, it has yet to be critically evaluated. This study aimed to compare behavioral and semen parameters in Catalan, Balearic, Amiata, and Miranda jacks treated with the PGF2α analogue cloprostenol sodium immediately prior to exposure to an estrus jenny. Nineteen donkeys were assigned in a crossover design to receive cloprostenol sodium (125µg, i.m.; n = 53 collections) or saline (1 mL, i.m.; n = 53 collections). There were no differences for erection (52/53 vs. 52/53) or ejaculation (52/53 vs. 48/53) for collection attempts assigned to saline or cloprostenol sodium, respectively. Cloprostenol sodium significantly hastened treatment-to-erection and treatment-to-ejaculation times from 12.0 ± 1.6 to 6.0 ± 1.6 min and from 14.0 ± 1.4 to 9.6 ± 1.4 min, respectively. Significant effects of breed and age were observed in behavioral and parameters, but there were no effects of cloprostenol sodium administration on semen parameters. In conclusion, cloprostenol sodium administration immediately prior to semen collection hastened time to collect semen in donkeys with no detrimental effects on semen quality and can be used by practitioners to circumvent long delays in donkey semen collection.

Age-related changes of seminiferous tubule morphology, interstitial fibrosis and spermatogenesis in dogs.

There are age-related changes in testicular anatomy and physiology whereby there are modifications of sperm production and reproductive hormone functions. Effects of age on testicular microanatomy are well documented in humans, while there is limited understanding of these changes in dogs. The aim of this study was to evaluate age-related changes of seminiferous tubule morphology, interstitial fibrosis and spermatogenesis in dogs. Dogs (n = 32) were divided into four age groups: peripubertal (n = eight), relatively younger (n = seven), reproductively mature (n = seven) and relatively older (n = ten). Picrosirius Red stained sections were used for morphometrical analysis of testicular tissues, while the characteristics of seminiferous epithelium were assessed using a modified Johnsen scoring system for haematoxylin and eosin stained sections. Seminiferous epithelium and seminiferous tubule area increased from peripuberty to reproductive maturity, indicating there were changes during sexual maturation and subsequently there were decreases with further aging. There was a similar age-related trend for changes in seminiferous epithelium height with values being greatest in reproductively mature dogs; while there were no age-related differences in tubular diameter. Collagen content in the testicular interstitium gradually decreased from peripuberty to the age when dogs were reproductively mature and there were subsequent increases in relatively older dogs, thus, there was an association between the extent of testicular fibrosis and senescence. There was a decrease in spermatogenetic functions from relatively younger to older ages. Further investigations are warranted to establish mechanisms responsible for age-related changes of testicular morphology and related clinical implications.

Embryo technologies in donkeys (Equus Asinus).

In industrialized countries, the donkey population had a dramatic decrease during the last century and this has brought almost all European donkey breeds to risk. Embryo technologies have already been employed as a tool for the conservation of endangered equid species (e.g. Equus Przewalskii). Today it is possible to obtain pregnancies after the transfer of donkey embryos in synchronized recipients, even though embryo transfer is not widely used as a reproductive technique in this species. So far, very few foals are born after transfer of cryopreserved embryos. To date, no pregnancies have been reported from in vitro fertilization studies. The use of the donkey as a dairy animal, as well as the increased importance of the conservation of endangered donkey breeds, could be the engine for the development and spread of embryo technologies in this species. However, funding from national and international research agencies in this field is still scarce, especially in Europe.

Role of body condition score and adiponectin expression in the progression of canine mammary carcinomas.

Obesity has been identified as a risk factor for developing breast cancer in post-menopausal period in humans and has been suspected to be associated with a worse prognosis also in the bitch. The aims of this study were to investigate the association between body condition score (BCS) and the prognosis of canine mammary carcinomas (CMCs) and the relationships between adiponectin expression and tumour behaviour. Seventy-three bitches with tubular, tubulopapillary, solid or complex carcinomas were included in the present study. For each dog, evaluation of BCS was conducted using a nine-point BCS system and the study population was divided into normal weight (4-5/9 points; n = 42), overweight (6-7/9 points; n = 19) and obese (8-9/9 points; n = 12). Type of diet (commercial, homemade or mixed) was recorded. After surgical excision, histological type, tumour size and nodal status were assessed and adiponectin expression was determined and quantified by immunohistochemistry and morphometric analysis. CMC histotype was not correlated with BCS, while a positive correlation between BCS and histological grade (p < .01) was observed. Overweight and obese bitches combined showed a shorter cancer-specific survival than normal weighted bitches (p < .01). Bitches fed with a homemade diet had a higher BCS than dogs fed with a commercial one, although no relationship was observed between diet and  cancer-specific survival. Thirty-six CMCs scored positive for adiponectin expression (49%), but no correlation was found between the hormone expression and either CMC characteristics or prognosis. In conclusion, a higher BCS seems to be related with a higher prevalence of more aggressive CMCs and negatively affects the survival time in bitches with these mammary tumours.

Relationship between placental characteristics and puppies' birth weight in toy and small sized dog breeds.

Puppy neonatal mortality may be related to low birth weight, which has been shown in humans to be linked to placental factors. The relation between the newborn puppy and the placental characteristics has been poorly investigated in dogs. Twenty bitches, 9 toy-sized (i.e. < 5 Kg) and 11 small-sized (i.e. 5 to 10 Kg), were included in this study. During natural delivery or c-section, puppies were identified and their order of birth, sex and weight were recorded. Puppy weights were registered at birth and daily until Day 6. Placentas were weighed after removal of extraplacental adnexa, after which a photo was taken and morphometrically assessed. The Total Placental Area (TPA) and the Transfer Zone Area (TZA) were calculated and their surface expressed in mm2. Immunohistochemistry with monoclonal mouse anti CD31 antibody was used to identify fetal and maternal vessels in the placental labyrinth zone. A vascularization index (VI) was determined for each placenta and the Total Vascular Area (TVA) was estimated. Puppies' birth weight correlated positively with placental weight (P < 0.001, r = 0.689). A positive correlation was found between the puppies' birth weight and TPA (P < 0.001, r = 0.786), TZA (P < 0.001, r = 0.772), and TVA (P < 0.001, r = 0.482). A positive correlation was also found between placental weight and TPA (P < 0.001, r = 0.661), TZA (P < 0.001, r = 0.583), and TVA (P < 0.001, r = 0.333). In the small-sized breeds, the placentas of low-weight puppies were lighter and had a smaller TZA and TVA (P < 0.05). The VI was higher in the placentas of the toy-sized compared to small-sized bitches (P < 0.01). No effect of parity, litter size, or sex of the puppy was observed on birth weight or growth rates on Day 6. The growth pattern of low-weight puppies did not differ from that of the other puppies during the first 6 days of life. As in humans, placental weight, the extension of the transfer zone and placental total vascular area correlates closely with the puppies' birth weight in normal pregnancies. Our data could represent reference values for placental weight, TZA, TVA and VI in toy and small-sized dog breeds.

Pharmacokinetics of levosulpiride after single-dose administration in goats (Capra hircus) by different routes of administration.

Levosulpiride (LSP) is the l-enantiomer of sulpiride, and LSP recently replacing sulpiride in several EU countries. Several studies about LSP in humans are present in the literature, but neither pharmacodynamic nor pharmacokinetic data of LSP is present for veterinary species. The aim of this study was to assess the pharmacokinetic profile of LSP after intravenous (IV), intramuscular (IM), and oral (PO) administration in goats. Animals (n = 6) were treated with 50 mg LSP by IV, IM, and PO routes according to a randomized cross-over design (3 × 3 Latin-square). Blood samples were collected prior and up to 24 hr after LSP administration and quantified using a validated HPLC method with fluorescence detection. IV and IM administration gave similar concentration versus time curve profiles. The IM mean bioavailability was 66.97%. After PO administration, the drug plasma concentrations were detectable only in the time range 1.5-4 hr, and the bioavailability (4.73%) was low. When the AUC was related to the administered dose in mg/kg, there was a good correlation in the IV and IM groups, but very low correlation for the PO route. In conclusion, the IM and IV administrations result in very similar plasma concentrations. Oral dosing of LSP in goats is probably not viable as its oral bioavailability was very low.