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There are no refereed reports on vitrification of domestic cat embryos derived from in vitro-matured oocytes and transferred using a nonsurgical embryo transfer technique. The aim of this study was to verify the effects of vitrification on the in vitroand in vivodevelopmental ability of in vitro-produced (IVP) cat blastocysts. Oocytes recovered from minced ovaries were matured, fertilized, and cultured in vitroas previously reported (Merlo B et al.2005 Theriogenology 63, 2032–2039). On Day 7 of in vitroculture (IVC), blastocysts were selected and vitrified in straws (Cristal ET 0.25 mL, 133 mm, IMV-Technologies, Paillette Crista, France). For vitrification (modified from Campos-Chillòn LF et al.2006 Theriogenology 65, 1200–1214), the embryos were transferred in 1 mL of V1 [ethylene glycol 3.5 min HEPES synthetic oviductal fluid (HSOF)] for 3 min, and then in 10 μL of V2 (ethylene glycol 7 m, galactose 0.5 m, Ficoll 70 18% in HSOF) for 20 s. Finally, the embryos were loaded in straws preloaded with 190 μL of dilution solution (galactose 0.5 min HSOF). Straws were heat sealed and immediately plunged into liquid nitrogen. Vitrified embryos were warmed in air for 10 s, and then in a waterbath at 37°C for 30 s. For developmental ability and in vitroevaluation, 27 embryos were warmed and immediately examined: 25 re-expanded, 2 did not re-expand, and 1 had damaged zona pellucida. Re-expanded embryos were cultured in SOF plus amino acids, 16 mg mL-1BSA, and 5% fetal bovine serum at 38.5°C in 5% O2, 5% CO2, 90% N2. After 24 h of IVC, only 4 blastocysts were expanded, and after 48 h, embryos were clearly degenerated or shrunk. in vivodevelopmental ability was tested by nonsurgical embryo transfer of 8 vitrified-warmed embryos and 6 IVP fresh embryos into 2 natural estrus queens, injected with 200 IU of hCG i.m. (Day 0) for induction of ovulation. Ovulation was confirmed by plasmatic progesterone assay on Day 5. Nonsurgical embryo transfer was made on Day 8 using the catheter proposed by Zambelli et al.2001 for transcervical insemination in the cat. The catheter was connected to a 1-mL syringe and loaded with the embryos. Then, it was inserted in the vagina and transrectally guided into the uterus, where the embryos were deposited. To assess pregnancy status, abdominal ultrasonography was done on recipients on Day 13, 25, and 40. On Day 13, an embryonic vesicle was observed in both queens, although a smaller diameter than expected was detected in the recipient of the vitrified embryos. On Day 25, a viable embryo was detected only in the recipient of fresh IVP embryos. On Day 40, the gestational chamber was still present but no sign of a viable embryo was detected. Further studies are in progress to improve the nominal incidence of pregnancy and frequency of embryo survival after vitrification. Nevertheless, the preliminary results obtained using an AI catheter for nonsurgical embryo transfer are encouraging, and the improvement of the technique could make it reliable in the cat.Supported by Animal Stem Cells Laboratory, Regione Emilia Romagna, PRRIITT Project Number M-404AIWTSV. [ABSTRACT FROM AUTHOR]

Hematopoietic neoplasms are common in dogs; however, their association with pregnancy has not been previously reported in veterinary medicine. This rare occurrence presents a variety of diagnostic, therapeutic, prognostic, and ethical challenges. We report a case of a 3-year-old pregnant Bernese Mountain Dog diagnosed with multicentric aggressive unclassified hematopoietic cancer associated with paraneoplastic hypercalcemia during pregnancy. The dog died 7 days after diagnosis, and at Day 36 of pregnancy before any treatment decision could be made. Post-mortem evaluation, including histology, immunohistochemistry, and clonality analysis, led to the diagnosis of an unclassified hematopoietic cancer affecting the uterus and placenta, with no evidence of fetal involvement. The placenta likely acted as a barrier, preventing neoplastic involvement of the fetuses. Alternatively, the pregnancy might have been too early for the hematopoietic neoplasm to affect the labyrinth zone of the placenta and the fetuses. The dramatic disease progression could be explained by compromised cell-mediated immunity during pregnancy. This immunodeficient state is induced by embryonic, maternal, and hormonal factors, which suppress the response to mitogens to prevent rejection of the placenta and the conceptuses. Thus, pregnant dogs might exhibit increased vulnerability to cancer and infectious diseases that rely on cell-mediated immunity for host defense., (© 2024 American Society for Veterinary Clinical Pathology.)

Objectives: Several anaesthetic protocols for caesarean section (c-section) have been described in dogs; however, anaesthesiological studies in cats undergoing c-section are lacking. The aim of this study was to assess the viability and outcome of kittens born from queens undergoing emergency c-section and receiving propofol or alfaxalone for anaesthetic induction and isoflurane for maintenance of general anaesthesia., Methods: All cats admitted to the Veterinary Teaching Hospital of the University of Bologna between January 2014 and December 2022 for dystocia that had undergone an emergency c-section were analysed in this retrospective study. The queens received propofol 2-6 mg/kg IV (group P) or alfaxalone 1-3 mg/kg IV (group A) administered slowly; general anaesthesia was maintained with isoflurane. Survival of all the kittens was evaluated at birth and after 24 h; in addition, heart rate, respiratory rate and mucous membrane colour were evaluated in the kittens at birth., Results: Of the 14 cats included in the study, eight received alfaxalone (group A) and six received propofol (group P) for induction of the general anaesthesia. A total of 50 kittens were born by c-section: 30 kittens in group A and 20 in group P. The overall survival of the kittens was 90% at birth: 96.7% (29/30) for group A and 80% (16/20) for group P. The 24 h survival rate was 93.1% for group A and 87.5% for group P., Conclusions and Relevance: The results of the present study demonstrated that in cats undergoing c-section, both alfaxalone and propofol are feasible for the induction of general anaesthesia., Competing Interests: Conflict of interestThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Assisted reproduction techniques play a significant role in veterinary medicine, and although they are widely used in domestic animals, they are also becoming increasingly relevant in clinical practice for wild felids, especially in the conservation efforts for endangered species. In this study, the result of two semen collection techniques, urethral catheterization after pharmacological induction (Ur.Ca.P.I.) and electroejaculation, are described, aiming to provide new practical information about sperm collection using the Ur.Ca.P.I. technique and electroejaculation in tigers and leopards, describing the authors' experience and presenting new data and observations. The following descriptive study included two subjects of Panthera tigris species and two of Panthera pardus . These subjects, after general anesthesia, underwent sperm collection initially with Ur.Ca.P.I. and, subsequently, with electroejaculation. Sampling was made possible in both species thanks to the use of electroejaculation. Sperm volumes in leopards ranged from 0.3 to 0.5 mL and in tigers from 0.5 to 2.177 mL. Sperm concentration in leopards ranged from 136 × 10 6 to 280 × 10 6 sperm/mL, and in tigers, from 21.5 × 10 6 to 354 × 10 6 sperm/mL. Urethral catheterization gave positive results in leopards, with sperm volumes ranging from 25 up to 150 µL and a concentration ranging from 110 × 10 6 up to 1082 × 10 6 sperm/mL. In tigers, unlike in leopards, the use of the Ur.Ca.P.I. technique encountered difficulties that did not allow satisfactory results to be obtained. Therefore, it would be useful to test the feasibility of urethral catheterization on a larger group of individuals in order to have more meaningful feedback. Finally, because electroejaculation always allowed semen collection in tigers, with a higher sperm quality than samples collected by Ur.Ca.P.I., we currently consider it the technique of choice for the collection of semen material in this species.

This study presents an innovative method for synthesizing β-amino carbonylated compounds, specifically 2-[phenyl(phenylamino)methyl] cyclohexanone, achieving high conversions and diastereomeric ratios. Using trypsin or α-chymotrypsin in both free and immobilized forms on titanate nanotubes (NtsTi), synthesized through alkaline hydrothermal methods, successful immobilization yields were attained. Notably, α-chymotrypsin, when free, displayed a diastereoselective synthesis of the anti-isomer with 97 % conversion and 16 : 84 (syn : anti) diastereomeric ratio, which slightly decreased upon immobilization on NtsTi. Trypsin, in its free form, exhibited diastereoselective recognition of the syn-isomer, while immobilization on NtsTi (trypsin/NtsTi) led to an inversion of diastereomeric ratio. Both trypsin/NtsTi and α-chymotrypsin/NtsTi demonstrated significant catalytic efficiency over five cycles. In conclusion, NtsTi serves as an effective support for trypsin and α-chymotrypsin immobilization, presenting promising prospects for diastereoselective synthesis and potential industrial applications. Furthermore, it offers promising prospects for the diastereoselective synthesis of 2-[phenyl(phenylamino)methyl] cyclohexanone through multicomponent Mannich reaction and future industrial application., (© 2024 Wiley-VCH GmbH.)

The development of endoscopic transcervical catheterization (ETC) in the queen increases the interest in handling fresh and cryopreserved feline semen. The ETC requires a small volume of the insemination dose with a high concentration, not easily reached with the actual frozen technique in this species. Centrifugation is widely used to concentrate spermatozoa for several purposes, but this process is detrimental to spermatozoa. This study verified the effects of conventional and cushioned centrifugation on fresh and cryopreserved feline spermatozoa. To this, semen was collected from 20 toms, grouped in seven pools and diluted. After dilution, the pools were divided into two aliquots, the first used for centrifugation on fresh semen, and the second, after freezing, on cryopreserved semen. Centrifugation regimens were: conventional at 500×g, conventional at 1000×g, and cushioned (iodixanol) at 1000×g. The sperm recovery rate was calculated for the three centrifugation regimens, and sperm kinematics, membrane and acrosome integrity, and plasma membrane stability on viable spermatozoa were assessed as endpoints. The data reported in this study showed that the centrifugation at 500×g resulted in negligible effects on both fresh and cryopreserved spermatozoa, but the lower recovery rate (62.4 ± 3.1 % and 60.2 ± 1.6 %, respectively) underlines the loss of a large proportion of spermatozoa, unfavourable in a species with small total sperm ejaculated. On the other hand, the centrifugation at 1000×g improved the recovery rate (86.9 ± 4.3 % and 89.8 ± 2.4 % in fresh and cryopreserved samples, respectively), but was more deleterious for feline spermatozoa, especially in cryopreserved samples (i.e. total motility of 40.7 ± 5.4 % compared with 57.2 ± 9.8 % in cryopreserved uncentrifuged samples, P < 0.05), resulting in artificial insemination doses of lower quality. The recovery rate in cushioned centrifugation appeared less efficient, likely due to the small volume of feline samples, which makes difficult the separation of sperm pellet and cushioned fluid. Interestingly, in cryopreserved samples centrifuged at 1000×g the number of viable spermatozoa with membrane destabilization (31.3 ± 3.2 %) was greater than uncentrifuged (4.1 ± 0.7 %, P < 0.05) and those centrifuged at 500×g (9.8 ± 1.3 %, P < 0.05), suggesting modifications induced by the cryopreservation amplifies centrifugation sublethal damage on feline spermatozoa. Cushioned centrifugation on cryopreserved samples showed kinematics similar to uncentrifuged samples, but higher viable spermatozoa with membrane destabilization (37.4 ± 3.4 % vs 4.1 ± 0.7 %; P < 0.05). In felines, g-force is crucial for sperm recovery rate during centrifugation, with better results at 1000×g; on the other hand, greater g-forces could have a significant impact on the quality of feline insemination dose, especially in cryopreserved samples., Competing Interests: Declarations of competing interest None., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Data from three cases of uterus masculinus were retrieved from 2014 to 2018. Two out of three cases presented clinical signs compatible with systemic infection, as observed in bitches with pyometra. Ultrasound examination revealed a tubular fluid-filled structure with a thin irregular wall located cranially to the prostate and in continuity with the cranial part of the gland. In two cases, two other tubular fluid-filled structures were visualized in the caudal part of the abdominal cavity, ventrally to the prostate gland and urinary bladder. After surgical removal of these, histological examination revealed the presence of a uterine structure morphologically similar to the female counterpart. Various types of epithelial cell lining were found, including simple columnar, simple stratified and squamous epithelium associated with glands in the underlying stroma. Immunohistochemistry to anti-Müllerian hormone (AMH) produced a positive result on glands, while multifocal expression was found in the lining epithelium. AMH seems involved in the pathogenesis of uterus masculinus , but its role is not fully understood. Thorough clinical and ultrasonographical examinations, followed by a histological confirmation, are necessary to properly diagnose uterus masculinus in dogs.

For a long time, the main way to acquire the skills necessary for good veterinary practice has been the traditional apprenticeship model (observe, assist, and perform under supervision). However, in the last years, more creative and innovative teaching models have been adopted by academic institutions and, parallelly, the opportunities to gain hands-on experience for clinical and surgical procedures are becoming more limited. For that reason, the introduction of the models can provide a potential solution to the ethical and legal implications related to the use of live animals for educational purposes and the biosafety risks deriving from the manipulation of human/animal cadavers. The activity on this topic at DIMEVET and, in particular, the experience about teaching reproduction includes in-house production and use of models for learning skills both for degree (fifth and third years of course) and post-graduate courses. Our models are designed on the basis of two fundamental aspect: the student level and the teacher target. The aim of this study was to evaluate, for the first time in literature, how much the use of simulators impacts on practical teaching in degrees and post graduate courses at DIMEVET, considering different learners with different levels of knowledge and skills. Namely, effectiveness, adequacy and quality of simulators have been checked assessing students' learning experience and teacher's opinion by specific satisfaction questionnaires and considering the attendance at the training sessions and the results of a final examination (PHY group) and an in vivo test (POST group). The rate of satisfaction was high among the three groups considered and the participants that had used the models had a higher success, both in the final examination and in vivo test. Data collected have been useful in order to show an improvement in teaching at DIMEVET in the field of reproduction, despite the relatively short years of experience using simulation. With this study we demonstrate that the adequacy of the models is not correlated to how simulators is built, whether with basic materials or not faithfully in terms of anatomical appearance, but it is important that it is realistic in terms of psychomotor procedure. In conclusion, the use of models permits the student to gain the hand-eye coordination and dexterity necessary to perform certain skills and the models proposed are meeting the desired educational goals., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Elsevier Inc. All rights reserved.)

In small animal practice, prostatic diseases are increasingly encountered. All dogs may experience prostatic disease, but there should be particular attention to breeding dogs, as prostatic disease may decrease semen quality and fertility. The most common prostatic disease is benign prostatic hyperplasia (BPH), a non-cancerous enlargement of the gland affecting intact adult dogs, part of an aging process, including both an increase in cell numbers (hyperplasia) and in cell size (hypertrophy). Acute and chronic prostatitis, prostatic abscess, prostatic neoplasia and prostate squamous metaplasia also occur in dogs, in order of frequency. These diseases often lack pathognomonic clinical signs; therefore, a thorough clinical examination and a correct diagnostic protocol are essential to determine the most appropriate treatment or prophylaxis. Frequently dogs with BPH are asymptomatic, but when clinical signs are present, the most common is a serous to sanguineous urethral discharge. BPH therapy includes various options and is usually recommended when mild-severe signs are present or if clinical signs disturb the dog. In most dogs with this disorder, it is possible to maintain fertility by avoiding castration and choosing alternative therapeutic approaches., Competing Interests: Declarations of interest None., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Practical Relevance: Despite substantial advances in assisted reproductive techniques having been recently reported in cats, the use of these is limited and routine application is still far from being a reality in veterinary clinics. Nevertheless, there is an increasing demand from domestic cat breeders for artificial insemination (AI) techniques that are already commonly used in dogs. Where natural breeding is not possible in tom cats and queens of high breeding value, AI could offer a solution. Clinical challenges: AI in cats is more difficult than in other species - both in terms of semen collection/handling and oestrous cycle management given that ovulation must be induced., Aim: For practitioners wishing to perform AI in queens, there are challenges to overcome, and a good understanding of the techniques and procedures involved is pivotal. This review aims to contribute to improved knowledge by providing an overview of AI protocols, encompassing choice of breeding animals, procedures for semen collection, oestrus and ovulation induction, AI techniques and equipment., Equipment and Technical Skills: Depending on the animals involved and the specific AI technique chosen, essential equipment may include an artificial vagina, electroejaculator, endoscope (sialendoscope, which can be fairly expensive) and special catheters for transcervical insemination. Other instrumentation and materials needed are typically readily available in a veterinary clinic. In general, no particular skills are needed to perform the procedures described in this review, with the exception of endoscopic transcervical catheterisation, where the ability to use an endoscope is required., Evidence Base: The information and advice/recommendations provided are based on specific feline research and reviews published in scientific peer-reviewed journals, animal reproduction textbooks, and presentations at national and international congresses. The authors also drew on their own clinical experience with regard to the choice of protocols and procedures presented in this review.

Perineal hernia refers to the herniation of pelvic and abdominal viscera into the subcutaneous perineal region through a pelvic diaphragm weakness: a concomitant prostatic disease is observed in 25-59% of cases. Prostatectomy involves the removal of the prostate, either partially (partial prostatectomy) or completely (total prostatectomy). In case of complicated perineal hernia, staged procedures are recommended: celiotomy in order to perform colopexy, vasopexy, cystopexy, and/or to treat the prostatic disease, and perineal access in order to repair the perineal hernia. Very few reports relate prostatectomy using a perineal approach and, to the extent of the author's knowledge, this technique has not been thoroughly investigated in the literature. The aim of this article is to retrospectively describe the total perineal prostatectomy in dogs presenting perineal hernia with concomitant prostatic diseases which required the removal of the gland. The experience in six dogs (three dogs with the prostate within hernial contents and three dogs with intrapelvic prostate) is reported as well as advantages, disadvantages, and limitations of the surgical procedure. In the authors' clinical practice, total perineal prostatectomy has been a useful surgical approach to canine prostatic diseases, proven to be safe, well tolerated, and effective.

Total or partial vulvo-vaginectomy or vaginectomy are not routinely performed due to the complexity of the techniques and because they are considered radical treatments. Little information can be found in the literature, as the same technique is often named in a different way by different authors, confusing the reader. Therefore, the aim of this essay is to describe five different surgical techniques: partial vaginectomy, complete vaginectomy, partial vestibule-vaginectomy, vulvo-vestibule-vaginectomy and vulvo-vestibulectomy. All techniques are described on the basis of the correct identification of the anatomical nomenclature related to structures involved in surgery, in order to give a more precise and unambiguous description and execution of surgical techniques. Moreover, possible intraoperative and perioperative complications and the authors' clinical experience in 33 dogs are described. All techniques are well tolerated and could be curative in case of benign or malignant tumours that have not yet metastasized and palliative in other cases. Moreover, they are also useful for therapeutic purposes for chronic vaginitis, severe vaginal cysts or congenital abnormalities. It is our opinion that having five different available techniques to approach vaginal disease is useful to perform the best surgery according to the clinical findings, patient's characteristics, technique invasiveness and whether it is palliative or not.

We report an outbreak of canine distemper virus (CDV) among stone martens (Martes foina) in Italy. After being rescued in Northern Italy between April and June 2018, six subjects were kept in a wildlife and exotic animal rescue center in Bologna province. Subjects have been monitored for 15 months in captivity. Within this time-lapse, two subjects died, while among the remaining four, only one showed clinical symptoms referable to distemper. Surviving subjects have been regularly tested for CDV by means of reverse transcriptase-PCR from conjunctival and oropharyngeal swabs for eleven months. The identified viruses belonged to the Wildlife-Europe CDV genetic subgroup. Neutralizing antibodies were detected at the end of the eleven months, when all subjects tested reverse transcriptase-PCR negative. Our findings confirm the circulation of the Wildlife-Europe CDV genetic subgroup (Europe 1/South America 1 lineage) within the Italian wildlife, and improve knowledge on viral infection in stone martens., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Postoperative complications impact on early and long-term patients' outcome. Appropriate perioperative fluid management is pivotal in this context; however, the most effective perioperative fluid management is still unclear. The enhanced recovery after surgery pathways recommend a perioperative zero-balance, whereas recent findings suggest a more liberal approach could be beneficial. We conducted this trial to address the impact of restrictive vs. liberal fluid approaches on overall postoperative complications and mortality., Methods: Systematic review and meta-analysis, including randomised controlled trials (RCTs). We performed a systematic literature search using MEDLINE (via Ovid), EMBASE (via Ovid) and the Cochrane Controlled Clinical trials register databases, published from 1 January 2000 to 31 December 2019. We included RCTs enrolling adult patients undergoing elective abdominal surgery and comparing the use of restrictive/liberal approaches enrolling at least 15 patients in each subgroup. Studies involving cardiac, non-elective surgery, paediatric or obstetric surgeries were excluded., Results: After full-text examination, the metanalysis finally included 18 studies and 5567 patients randomised to restrictive (2786 patients; 50.0%) or liberal approaches (2780 patients; 50.0%). We found no difference in the occurrence of severe postoperative complications between restrictive and liberal subgroups [risk difference (95% CI) = 0.009 (- 0.02; 0.04); p value = 0.62; I 2 (95% CI) = 38.6% (0-66.9%)]. This result was confirmed also in the subgroup of five studies having a low overall risk of bias. The liberal approach was associated with lower overall renal major events, as compared to the restrictive [risk difference (95% CI) = 0.06 (0.02-0.09); p value  = 0.001]. We found no difference in either early (p value  = 0.33) or late (p value  = 0.22) postoperative mortality between restrictive and liberal subgroups CONCLUSIONS: In major abdominal elective surgery perioperative, the choice between liberal or restrictive approach did not affect overall major postoperative complications or mortality. In a subgroup analysis, a liberal as compared to a restrictive perioperative fluid policy was associated with lower overall complication renal major events, as compared to the restrictive., Trial Registration: CRD42020218059; Registration: February 2020, https://www.crd.york.ac.uk/prospero/display&#95;record.php?RecordID=218059 .

Placenta is essential for the development of the fetus, and its impaired function can lead to a negative outcome (i.e., neonatal mortality). In dogs, investigations on placenta histology and neonatal outcome in healthy bitches are lacking, and a contribution is provided in this study to emphasize the use of placenta histology in practice. Fifty-one placentas from 11 litters were collected during cesarean section, classified according to the litter size (large (L) or small (S)) and the outcome, this latter as healthy (Group 1) or dead within 7 days (Group 2). The placenta/puppy weight ratio (PPR) was calculated, and specimens were formalin-fixed and paraffin-wax embedded, and on the resulting histological slides, capillary density (CD) was quantified. Among necrosis, calcification, and intravascular leucocytes, only the presence of multifocal-confluent necrosis (significantly more frequent in Group 2) was associated with a higher risk of death within 7 days (odds ratio = 30.7). Mixed logistic regression ruled out the effect on death both of a bitch and cesarean type (programmed vs. emergency). PPR and CD values were associated with litter size; large litters had lower PPR ( p < 0.01) and higher CD ( p < 0.05) than small litters. The relationship between PPR and CD was negative and significant ( p < 0.01). Necrosis was a frequent finding in canine placentas, but only when multifocal-confluent was it associated with a poor outcome. The litter size influenced PPR (lower in L) and CD (higher in L), and this is likely due to the plasticity of placenta adaptation.

Background: Appropriate perioperative fluid management is of pivotal importance to reduce postoperative complications, which impact on early and long-term patient outcome. The so-called perioperative goal-directed therapy (GDT) approach aims at customizing perioperative fluid management on the individual patients' hemodynamic response. Whether or not the overall amount of perioperative volume infused in the context of GDT could influence postoperative surgical outcomes is unclear., Methods: We conducted a systematic review and meta-analysis of randomized controlled trials (RCTs) comparing the efficacy of GDT approach between study population and control group in reducing postoperative complications and perioperative mortality, using MEDLINE, EMBASE and the Cochrane Controlled Clinical trials register. The enrolled studies were grouped considering the amount infused intraoperatively and during the first 24 h after the admission in the critical care unit (perioperative fluid)., Results: The metanalysis included 21 RCTs enrolling 2729 patients with a median amount of perioperative fluid infusion of 4500 ml. In the studies reporting an overall amount below or above this threshold, the differences in postoperative complications were not statically significant between controls and GDT subgroup [43.4% vs. 34.2%, p value = 0.23 and 54.8% vs. 39.8%; p value = 0.09, respectively]. Overall, GDT reduced the overall rate of postoperative complications, as compared to controls [pooled risk difference (95% CI) = - 0.10 (- 0.14, - 0.07); Chi 2  = 30.97; p value < 0.0001], but not to a reduction of perioperative mortality [pooled risk difference (95%CI) = - 0.016 (- 0.0334; 0.0014); p value = 0.07]. Considering the rate of organ-related postoperative events, GDT did not reduce neither renal (p value = 0.52) nor cardiovascular (p value = 0.86) or pulmonary (p value = 0.14) or neurological (p value = 0.44) or infective (p value = 0.12) complications., Conclusions: Irrespectively to the amount of perioperative fluid administered, GDT strategy reduces postoperative complications, but not perioperative mortality., Trial Registration: CRD42020168866; Registration: February 2020 https://www.crd.york.ac.uk/prospero/display&#95;record.php?RecordID=168866.

Introduction: This study analyzes the usefulness of the CHA2DS2-VASc score for mortality prediction in patients with acute coronary syndromes (ACSs) and evaluates if the addition of renal functional status could improve its predictive accuracy., Methods: CHA2DS2-VASc score was calculated by using both the original scoring system and adding renal functional status using 3 alternative renal dysfunction definitions (CHA2DS2-VASc-R1: eGFR <60 mL/min/1.73 mq = 1 point; CHA2DS2-VASc-R2: eGFR <60 mL/min/1.73 mq = 2 points; and CHA2DS2-VASc-R3: eGFR <60 mL/min/1.73 mq = 1 point, <30 mL/min/1.73 mq = 2 points). Inhospital mortality (IHM) and post-discharge mortality (PDM) were recorded, and discrimination of the various risk models was evaluated. Finally, the net reclassification index (NRI) was calculated to compare the mortality risk classification of the modified risk models with that of the original score., Results: Nine hundred and eight ACS patients (median age 68 years, 30% female, 51% ST-elevation) composed the study population. Of the 871 patients discharged, 865 (99%) completed a 12-month follow-up. The IHM rate was 4.1%. The CHA2DS2-VASc score demonstrated a good discriminative performance for IHM (C-statistic 0.75). Although all the eGFR-modified risk models showed higher C-statistics than the original model, a statistically significant difference was observed only for CHA2DS2-VASc-R3. The PDM rate was 4.5%. The CHA2DS2-VASc C-statistic for PDM was 0.75, and all the modified risk models showed significantly higher C-statistics values than the original model. The NRI analysis showed similar results., Conclusions: CHA2DS2-VASc score demonstrated a good predictive accuracy for IHM and PDM in ACS patients. The addition of renal dysfunction to the original score has the potential to improve identification of patients at the risk of death., (© 2021 S. Karger AG, Basel.)

High dose medetomidine 0.13 mg/kg can be used for semen collection in cats with variable results in terms of quantity and quality. Therefore, a variation in terms of distribution and elimination among patients has been hypothesised. The aim of the study was to characterise the pharmacokinetics of medetomidine (0.13 mg/kg) administered intramuscularly (IM) in healthy male cats. Eighteen male cats undergoing castration were included, and medetomidine (0.13 mg/kg) was administered IM. Venous blood samples were collected at 20, 30, 40, 50, 60, 75 and 90 minutes after medetomidine administration. Before orchiectomy, at T20, sperm collection was attempted. Plasma medetomidine concentrations were determined by liquid chromatography/mass spectrometry analysis. Semen collection was successful in 15/18 cats. The medetomidine plasma concentration following the IM administration of a bolus was best described using a non-compartment model. Time of maximum concentration was observed at 40 minutes (range 20-90); maximum concentration was 32.8 ng/mL (range 26.8-51.2). The median apparent clearance was 11.9 mL/kg/minute (range 0.7-43.8). In conclusion, medetomidine administered IM at 0.13 mg/kg reached its peak plasma concentration slowly and with variability among patients. In addition, it was characterised by low total body clearance probably due to the cardiovascular alterations associated with medetomidine administration.

Egg yolk (EY) is conventionally used to reduce sperm cryodamage, however, there has not be evaluation of whether there is a dose-dependent effect with inclusion of EY in semen extender. To enhance the knowledge about the protective effect of EY during cryopreservation of dog semen, a specific study was designed to evaluate the dose-dependent protection of the EY against osmotic and cryogenic damage of dog sperm. In the first experiment, sperm stored in an extender that contained graded EY concentrations (0 %, 5 %, 10 %, and 20 %) were diluted with hypo- or hyper-osmotic solutions (final osmolality of 75, 150, 300, 500, 1000 mOsm/kg). Results from sperm kinetic, membrane integrity (MI), mitochondrial activity, and normal morphology evaluations indicated osmotic stress has especially marked effects on the kinetic capacity of spermatozoa, however, there were no direct effects on mitochondrial activity. In both hypo- and hyper-osmotic conditions, EY had a protective effect regardless of concentration. In the second experiment, semen samples were diluted in extenders at increasing EY concentrations (0 %, 5 %, 10 %, and 20 %) and cryopreserved. Effects on sperm kinetics, membrane and acrosome integrity and mitochondrial membrane potential indicated there was improved sperm viability after thawing when the EY concentration was 5 % and 10 %, and lesser viability when it was 20 %. These results indicate, for the first time, that EY reduces osmotic and cryogenic damage when used at 5 % or 10 % concentrations, and that these concentrations can be used to protect dog spermatozoa more effectively than the conventionally used concentration (20 %)., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In small animal practice, prostatic diseases are increasingly encountered. All dogs may experience prostatic disease, but particular care should be addressed to breeding dogs, in which prostatic affection may lead to decrease in semen quality and fertility. The most common prostatic disease is the benign prostatic hyperplasia (BPH) followed by prostatitis, prostatic neoplasia and prostate squamous metaplasia. These diseases do not have pathognomonic symptoms, therefore, making a correct diagnosis may not be easy. An accurate clinical examination and a correct diagnostic protocol are essential in order to begin the most appropriate treatment, and also to do a good prophylaxis where it is possible. BPH therapy is usually recommended when mild-severe signs are present or if symptoms disturb the patient. New therapeutic approaches, both medical and surgical, allow to maintain fertility in most animals with prostatic disorders. Prostate cancer is relatively infrequent. Elective therapy is the surgical one, but it is considered palliative and can result in important post-operative complications. The aim of this paper is to lay down the most appropriate diagnostic process describing the aetiologies of prostatic disease, their symptoms, the right investigative tools and therapy., (© 2019 Blackwell Verlag GmbH.)

Objective: To measure plasma methadone concentrations in bitches and the umbilical cords of their puppies after systemic or epidural administration., Study Design: Prospective, randomized, clinical study., Animals: A total of 27 healthy pregnant female dogs undergoing caesarean section, 4.3 ± 2.3 years of age and weighing 19.9 ± 13.2 kg., Methods: The dogs were randomly divided into three groups: 1) intramuscular methadone (0.3 mg kg -1 ) (group MET; n = 9); 2) epidural methadone (0.1 mg kg -1 ) (group METEPI; n = 9); and 3) epidural lidocaine (4.4 mg kg -1 ) [group CON (control group); n = 9]. Ten minutes before induction, methadone was administered intramuscularly to the group MET dogs. Anaesthesia was induced with propofol and maintained with isoflurane. Cardiovascular and respiratory parameters were monitored throughout the anaesthesia. After induction, epidural anaesthesia was administered to dogs in groups METEPI and CON. Before any treatment (T0) and, as soon as the last foetus was removed from the uterus (T1), venous blood samples were collected from each dog into heparinized tubes; the umbilical cords were collected and stored at -80 °C until pharmacological analysis was carried out. The samples were analysed using ultra performance liquid chromatography., Results: The cardiorespiratory parameters of the bitches and of the puppies at birth, and the Apgar scores did not differ significantly between groups. At T1 both the median maternal methadone plasma concentration and the median methadone umbilical cord concentration were higher in group MET compared to group METEPI (p = 0.0018 and p = 0.004, respectively). The maternal plasma concentration was higher than the concentration in the umbilical cords (p = 0.05) in group METEPI but not in group MET (p = 0.25)., Conclusions and Clinical Relevance: Epidural methadone (0.1 mg kg -1 ) administered to bitches undergoing caesarean section is associated with lower umbilical cord methadone concentrations as compared with intramuscularly administered methadone at higher dosages (0.3 mg kg -1 )., (Copyright © 2018 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Dog sperm cryopreservation is gaining importance both in breeding dogs for commercial purposes and for pet animals. Anyway, cryopreservation of mammalian spermatozoa, including dog ones, induces some negative effect on sperm fertility, leading to a lower use of this technique and limiting its widespread use. Therefore, studies to improve the quality of canine semen after cryopreservation could have a relevant impact on both the scientific advancement and the clinical practice. The aim of the present work was to investigate the putative ameliorative effect of Epigallochatechin-3-gallate (EGCG) addition to post thawing medium on dog sperm motility, mitochondrial activity, acrosome integrity and on zona-binding ability (zona binding assay). Spermatozoa were thawed in Tris-fructose-citrate medium supplemented with EGCG (0, 25 and 50 μM) and sperm motility, mitochondrial activity and acrosome integrity were assayed at 0.5, 1.5, 3 and 6 h after post thawing incubation at 37 °C. An aliquot of semen from each treatment group after 1.5 h post thawing incubation was washed and used to perform heterologous (using porcine oocytes) or homologous zona binding assay. The results obtained showed that no significant effect is exerted by EGCG on sperm parameters analysed neither at 0.5, 1.5, 3 or 6 h after thawing excepting for the reduction of the percentage of live cells with active mitochondria at the higher dose at 6 h; furthermore, both homologous or heterologous zona binding ability, was not influenced by EGCG. In conclusion, EGCG supplementation to thawing medium does not improve dog sperm quality or zona binding capacity., (Copyright © 2018 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)

Objective: To investigate the pharmacokinetics of buprenorphine and its main active metabolite, norbuprenorphine, after administration of an intravenous loading dose followed by constant rate infusion (CRI) in dogs., Study Design: Prospective, clinical study., Animals: A total of seven healthy dogs undergoing elective ovariectomy., Methods: Buprenorphine was administered as a loading dose (intravenous bolus of 15 μg kg -1 ) followed by CRI (2.5 μg kg -1 hour -1 for 6 hours). Moreover, intraoperative analgesia was supplemented by an intramuscular carprofen (4 mg kg -1 ) injection, administered prior to surgery, and by lidocaine, administrated through subcutaneous infiltration and through a splash on the ovarian vascular pedicle during surgery. Pain and sedation were scored for all animals throughout the 24-hour study period and rescue analgesia was administered when a visual analogue scale score was > 40 mm. Blood samples were collected from a jugular catheter at regular intervals, and plasma concentrations of buprenorphine and norbuprenorphine were determined by a validated liquid chromatography-tandem mass spectrometry method., Results: Buprenorphine showed a two-compartment kinetic profile. Maximum concentration was 23.92 ± 8.64 ng mL -1 at 1 minute (maximum time); elimination half-life was 41.87 ± 17.35 minutes; area under the curve was 486.68 ± 125.66 minutes ng -1 mL -1 ; clearance was 33.61 ± 13.01 mL minute -1 kg -1 , and volume of distribution at steady state was 1.77 ± 0.50 L kg -1 . In no case was rescue analgesia required. Norbuprenorphine resulted below the lower limit of quantification in almost all samples., Conclusions and Clinical Relevance: The results suggest that a buprenorphine CRI can be a useful tool for providing analgesia in postoperative patients, considering its minor side effects and the advantages of a CRI compared to frequent boluses. The negligible contribution of norbuprenorphine to the therapeutic effect was confirmed., (Copyright © 2017 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Blood lactate concentration is known to be a good prognostic indicator associated with the severity of illness and the patient's outcome both in human and veterinary medicine. It also plays a significant role in the assessment of the newborn, being a good indicator of fetal hypoxia and the ideal predictor of morbidity at term in babies. In veterinary neonatal medicine, hyperlactatemia is considered a valid prognostic marker in critically ill foals; moreover, blood lactate measurement has been proposed for the evaluation of newborn viability and the assessment of fetal distress during delivery in dogs. Unfortunately, only a few studies have been published concerning the canine species. The present work examines 67 brachycephalic newborn dogs and their mothers, with the aim to evaluate the time-dependent changes of blood lactate and glucose concentration during the first 24 h after vaginal or caesarean delivery both in puppies and bitches. To our knowledge, this is the first published study examining the time-dependent changes of these parameters in the bitch after parturition. Within the studied population of puppies, non-surviving was significantly associated with a higher lactatemia and a lower APGAR score. Blood lactate was high at birth then progressively decreased during the first 24 h of life and a lack of normalization of blood lactate levels within this time interval was suggestive for a poor prognosis for the newborn dogs; moreover, the decrease appeared to be slower after vaginal delivery. Lactatemia also showed a positive correlation with glycemia at birth. Concerning the bitches examined, blood lactate was found to be significantly higher after vaginal delivery than after caesarean section; the normalization occurred within 24 h after parturition. Blood glucose level was significantly higher at 2 h from delivery both in the group of bitches submitted to caesarean section and in those undergoing natural whelping but no statistical correlation was found between maternal glycemia and lactatemia. The results of the present study highlighted that the monitoring of lactatemia during the first 24 h of life, in association with the assessment of the APGAR score at birth, can be an useful prognostic tool helping to identify the most severely distressed puppies and to provide them an adequate support., (Copyright © 2017. Published by Elsevier Inc.)

Objectives: This study aimed to assess non-invasively the cardiovascular effects of high-dose medetomidine on healthy male cats undergoing semen collection., Methods: Haemodynamic evaluations were assessed on the basis of clinical examination, systolic arterial pressure (SAP) and transthoracic echocardiographic examination. Eight client owned, male domestic shorthair cats were sedated with a bolus of medetomidine intramuscularly (IM; 0.13 mg/kg), and semen collection was performed. A second transthoracic echocardiographic examination and SAP measurement were carried out 15 mins after sedation. At the end of the examination, the patients received a bolus of atipamezole (0.3 mg/kg) IM., Results: The cats were deeply sedated, relaxed and laterally recumbent during the entire procedure. No rhythm abnormalities were observed during the examinations and no significant increase in SAP was recorded. Heart rate dropped from 200 ± 33 to 92 ± 13.1 beats per min after sedation. There was a significant increase in left ventricular dimensions and the left atrial area. The parameters of left ventricular systolic function were reduced, as were systemic and pulmonary cardiac outputs. Peak diastolic wave velocities were significantly reduced, while isovolumic contraction and relaxation time of the left ventricle were prolonged. Aortic valve insufficiency was recorded for all cats, while mitral valve insufficiency was noted in five cats. None of the subjects developed systolic anterior motion of the mitral valve., Conclusions and Relevance: The protocol allowed us to collect good semen samples in healthy cats. However, high-dose medetomidine induces significant haemodynamic effects on the feline heart, mainly due to a reduced heart rate, an increased cardiac preload and impaired systolic function. The animals recovered from the anaesthesia, after antagonist administration, without showing any clinically relevant consequences., (© ISFM and AAFP 2015.)

The aim of this study was to evaluate the influence that different protocols of urethral catheterization after pharmacological induction (Ur.Ca.P.I.) may have on the semen quality of the domestic cat. The study has been divided into two experiments: one in which different dosages of medetomidine administrated are evaluated and the second one in which the timing of the catheterization after pharmacological induction is tested. In the first experiment, 18 cats were sedated with the recommended dosage of medetomidine (130 μg/kg i.m.) while the other 18 were sedated with a lower dose of the same drug (50 μg/kg i.m.). In the second experiment, three groups were implemented, each containing 25 subjects. In group 1, the semen collection was performed immediately once the pharmacological effect of the drug was reached; in group 2, the semen collection was performed three times every 5 min after the pharmacological effect was reached; finally, in group 3, Ur.Ca.P.I. was performed 20 min after the pharmacological effect was reached. All the different protocols permitted sperm collection, nevertheless the first experiment showed a better quality in terms of volume, concentration, total number of spermatozoa (p < 0.01) and quality of the movement (motility p < 0.05 and forward progressive motility p < 0.01), using a high medetomidine dosage rather than 50 μg/kg i.m. In the second experiment, forward motility was statistically higher (p < 0.01) in the first group and total volume was higher (p < 0.01) in the second and third group, while other parameters were statistically not different. Results suggest that a single catheterization immediately after the onset of the pharmacological effect leads to a good-quality semen with the lowest possibility of damaging the urethra and that a sedation with 130 μg/kg of medetomidine leads to a better quality sperm collection than 50 μg/kg does., (© 2015 Blackwell Verlag GmbH.)

The present study consists of two distinct parts, experiment 1 and experiment 2. In experiment 1, 13 anestrous queens were treated with a 4.7-mg deslorelin subcutaneous implant to assess its effectiveness in inducing estrus in the domestic cat. Deslorelin is currently used for the reversible suppression of ovarian and testicular activity in dogs and cats and for estrus induction in the bitch. Estrus induction is also reported in the queen but never reported with a targeted study. All the queens showed a positive response to the induction protocol, and estrus was detected within an average of 5.0 ± 2.2 days after the implant placement in 13 out of 13 subjects (100%). Seven of 13 queens exhibited behavioral manifestations of estrus, and the mean number of follicles detected at ultrasound examination was 4.8 ± 1.6 per subject. In experiment 2, three of the queens previously treated with deslorelin for estrus induction were submitted to artificial insemination through endoscopic transcervical catheterization, a new nonsurgical technique for intrauterine sperm deposition. All of them (100%) were pregnant after insemination and they gave birth to healthy litters. The study, as a whole, proves the effectiveness of the 4.7-mg deslorelin subcutaneous implants in inducing estrus in the domestic cat and is, to our knowledge, the first study assessing fertility of the induced estruses. Moreover, it shows the effectiveness of endoscopic transcervical catheterization for artificial insemination in the queen., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The aim of this study was to define a protocol to store dog sperm before and after sorting to obtain an insemination dose sufficient to allow the conception by artificial insemination. Experiment 1 and 2 were performed to evaluate the more appropriate extender for preserving at room temperature dog sperm before and after sorting. Four extenders were tested: (1) Tris-fructose-citrate (TFC), (2) Tris-glucose-citrate (TGC), (3) modified Tyrode's albumin lactate pyruvate medium (mTALP), and (4) third fraction of the ejaculate (after centrifugation at 5000× g for 10 minutes; III FRAC). Experiment 3 and 4 were performed to evaluate the ability of dog semen to withstand sex sorting and freezing/thawing. Modified Tyrode's albumin lactate pyruvate medium was the best extender for canine sperm storage at room temperature (20 °C-25 °C) before (total motility: TFC, 8.3 ± 1.7; TGC, 50.0 ± 11.5; mTALP, 70.0 ± 0.1; III FRAC, 25.0 ± 1 0.4; P < 0.05) and after sorting (total motility: TFC, 7.3 ± 1.5; TGC, 10.3 ± 1.5; mTALP, 33.3 ± 6.7; III FRAC, 8.7 ± 5.8; P < 0.05), even if at 24-hour sorted sperm quality was impaired in all extenders tested herein. Sperm quality decreased after sorting (total motility: control, 92.5 ± 0.9; sorted, 52.9 ± 6.0; P < 0.05) and, especially, after freezing/thawing (total motility: frozen control, 25.7 ± 4.1; frozen sorted, 2.4 ± 1.2; P < 0.05). In conclusion, mTALP is an appropriate medium for canine sperm storage before and soon after sorting (hours), but a long storage period of sexed sperm at room temperature is not adequate. Cryopreservation greatly impaired sperm quality, and further studies are needed to optimize the freezing protocol for sexed dog sperm., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Feline-assisted reproduction is still not routinely performed in veterinary practice, although there is an increasing interest on the subject by cat breeders. In recent years, many techniques for artificial insemination in the domestic cat have been developed with regard to the intrauterine deposition of sperm through the catheterization of the cervix. Transcervical catheterization has been described also for diagnostic and therapeutic purposes. This article provides the first description of a new method for cervical catheterization, under the direct visualization of the cervix, using a rigid endoscope and a new specially designed transcervical catheter. The procedure was performed on 14 queens with a success rate of 85.71%., (© 2014 Blackwell Verlag GmbH.)

The discovery of heme-induced respiration in Lactococcus lactis has radically improved the industrial processes used for the biomass production of this species. Here, we show that inhibition of the lactate dehydrogenase activity of L. lactis during growth under respiration-permissive conditions can stimulate aerobic respiration, thereby increasing not only growth efficiency but also the robustness of this organism.

Benign prostatic hyperplasia (BPH) is a spontaneous and age-related condition in humans and intact male dogs. A symptom index for BPH in men was created by the American Urological Association. In this study, it has been developed and statistically validated as a model to assign an objective score to canine BPH severity based on clinical signs observed and/or subjectively reported to the veterinarian by dog owners. The medical records of the Animal Reproduction Unit of University of Bologna (Italy) were used to select dogs with a clinical diagnosis of BPH. A data set was built up, and the animals were included in the statistical analysis as dependent variables. A score of 1-3 was assigned to the disease severity of each case based on signs annotated, graded using a scale ranging from 1 to 4. Signs of BHP were entered as predictors while disease severity as dependent variable to generate the predictive model. The model was finally used to re-classify each case of the data set, and the percentage of corrected predictions calculated. Overall, 373 subjects were entered in the model. Between them, 243, 107 and 23 animals have been represented based on medical records with a BPH severity score of 1, 2 and 3, respectively. The model correctly predicted the response variable in 97.3% of the cases. In this study, a BPH symptom index was created for the first time in dogs, which may be useful to standardize BPH severity with an objective score and to evaluate the necessity, the kind and the effectiveness of treatment., (© 2012 Blackwell Verlag GmbH.)