Short- and Long-Term Storage of Non-Domesticated European Mouflon (Ovis aries musimon) Cumulus-Oocyte Complexes Recovered in Field Conditions.

Reproductive biotechnologies can be used as a supporting tool, through gamete conservation and in vitro embryo production, in the preservation of invaluable and irreplaceable animal genetic resources. In the present study, immature mouflon cumulus-oocyte complexes (COCs) collected from ovariectomized female ovaries underwent short- or long-term conservation (24 h maintained in Earle's/Hank's (EH) medium or vitrification) under field conditions and afterwards transported to the laboratory where they were cultured for in vitro maturation (IVM) and assessed for oocyte meiotic competence and bioenergetic-oxidative status. Utilization of both storage techniques led to COC morphology preservation, as well as cumulus expansion and oocyte meiotic resumption after the IVM procedure. Quantitative bioenergetic-oxidative parameters were reduced in vitrified oocytes compared with EH ones. Immature COC storage needs to be optimized in both domesticated and non-domesticated sheep as a part of the strategy to avoid the loss of valuable genotypes of these animal species.

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.

The Effect of Cushioned Centrifugation, with and without Enzymatic Reduction of Viscosity, on the Motility Pattern and Kinematic Parameters of Dromedary Camel Bull Spermatozoa.

In order to contribute to the development of semen processing procedures in camelids, the aims of the present study were to evaluate (i) the effect of 35% seminal plasma incubation on dromedary camel epididymal sperm motility and kinematic parameters, (ii) the effects of centrifugation, with cushion fluid and enzymatic reduction of viscosity (Papain + E64) during ejaculate processing, on the motility and kinematic parameters of dromedary camel ejaculates. The incubation with seminal plasma significantly reduced the percentage of progressively motile spermatozoa as well as the proportion of medium progressive spermatozoa whilst increasing the percentage of non-progressive spermatozoa. The centrifugation procedure improved the sperms' kinematic parameters, and the highest values were observed for samples centrifugated with cushion fluid. The samples treated with Papain + E64 showed a significant increase in both total and medium progressive spermatozoa, along with a reduction of non-progressive spermatozoa (p < 0.05). The results of this investigation show that a simple, cheap, and effective procedure, such as cushioned centrifugation, could improve the motility patterns of dromedary camel spermatozoa; in combination with enzymatic reduction of viscosity, this method leads to the best results in terms of recovery rates and sperms' kinematic parameters.

New Strategies for Conservation of Gentile di Puglia Sheep Breed, an Autochthonous Capital of Millennial Tradition in Southern Italy.

Gentile di Puglia (GdP) is an autochthonous sheep breed of Southern Italy included among ovine breeds threatened by genetic erosion and extinction risk, which have been given attention by local and international institutions, thus emphasizing the need for germplasm conservation actions. In the present study, two assisted reproduction approaches, finalized for GdP conservation, were performed: (1) on-farm reproductive efficiency evaluation, expressed as pregnancy rate (PR), twin pregnancy rate (tPR), and body condition score (BCS), for three consecutive breeding cycles and (2) pre-pubertal lambs' immature cumulus-oocyte complex (COC) retrieval, vitrification, in vitro maturation (IVM), and assessment of meiotic stage and bioenergetic-oxidative status compared with those of other Italian and European commercial breeds. PR and tPR were progressively reduced over time. In all clinical examination times, BCS was significantly lower in nonpregnant ewes compared with pregnant ones. Fresh GdP pre-pubertal lamb COCs achieved meiotic maturation and showed healthy bioenergetic-oxidative status after IVM. Vitrification reduced the oocyte maturation rate in all groups. However, mature oocytes retained their cytoplasmic maturity, expressed as a mitochondria distribution pattern and activity, indicating promising developmental competence. In conclusion, clinical- and biotechnological-assisted reproduction approaches can support conservation strategies of GdP and other local sheep breeds in Southern Italy.

Update on canine anorchia: A review.

Abnormalities of the external genitals are an important issue in dog breeding because of the unfavourable qualities and characteristics of breeds, resulting in consistent economic losses. Despite their significance, little scientific attention has been given to these problems. Although there are several reviews on cryptorchidism in dogs, none have described anorchia. Testicular agenesis is a rare reproductive disorder with a congenital origin. Moreover, no author has described the diagnostic procedure for making a definitive diagnosis of anorchia in dogs. It is important to have a well-structured diagnostic scheme to help practical veterinarians make a confirmatory diagnosis. This review article aims to provide an update on canine anorchia diagnosis based on the poor research studies published in recent years. We have also contributed to the pathogenesis of this disorder using human medicine studies. Finally, the review includes therapeutic hypotheses that can be expanded in future studies.

Investigating and Modelling an Engineered Millifluidic In Vitro Oocyte Maturation System Reproducing the Physiological Ovary Environment in the Sheep Model.

In conventional assisted reproductive technologies (ARTs), oocytes are in vitro cultured in static conditions. Instead, dynamic systems could better mimic the physiological in vivo environment. In this study, a millifluidic in vitro oocyte maturation (mIVM) system, in a transparent bioreactor integrated with 3D printed supports, was investigated and modeled thanks to computational fluid dynamic (CFD) and oxygen convection-reaction-diffusion (CRD) models. Cumulus-oocyte complexes (COCs) from slaughtered lambs were cultured for 24 h under static (controls) or dynamic IVM in absence (native) or presence of 3D-printed devices with different shapes and assembly modes, with/without alginate filling. Nuclear chromatin configuration, mitochondria distribution patterns, and activity of in vitro matured oocytes were assessed. The native dynamic mIVM significantly reduced the maturation rate compared to the static group (p < 0.001) and metaphase II (MII) oocytes showed impaired mitochondria distribution (p < 0.05) and activity (p < 0.001). When COCs were included in a combination of concave+ring support, particularly with alginate filling, oocyte maturation and mitochondria pattern were preserved, and bioenergetic/oxidative status was improved (p < 0.05) compared to controls. Results were supported by computational models demonstrating that, in mIVM in biocompatible inserts, COCs were protected from shear stresses while ensuring physiological oxygen diffusion replicating the one occurring in vivo from capillaries.

The role of bicarbonate in the modulation of capacitation, spontaneous acrosome reaction and motility of equine fresh and frozen spermatozoa.

In this study, we defined the composition of the culture medium that yield a significant percentage of alive and functional equine spermatozoa during enough time before artificial insemination. The effects of sodium bicarbonate were analyzed in fresh and frozen semen in respect to sperm viability, capacitation, spontaneous acrosome reaction and several kinetic parameters such as total motility, progressive motility, VCL, VSL, ALH, BCF, LIN. Moreover, employing Bayk-6844 and Nifedipine, the involvement of L-type voltage-gated calcium channels in the modulation of intracellular calcium concentrations was investigated. Results evidenced an immediate effect of sodium bicarbonate in reducing fresh sperm viability and in increasing capacitation and spontaneous acrosome reaction of fresh and frozen spermatozoa. Furthermore, it affected total and progressive motility of fresh and frozen semen. Because of the sudden effects of the compound, we think that a buffer lacking sodium bicarbonate is more suitable to preserve the viability and the functional state of equine spermatozoa required to undergo at the right time those modifications necessary for fertilization. We also demonstrated that intracellular calcium modifications induced by Bayk-8644 and Nifedipine are not involved in signals related to vitality, capacitation, spontaneous acrosome reaction and motility.

The ultrasound-guided funicular block in cats undergoing orchiectomy: ropivacaine injection into the spermatic cord to improve intra and postoperative analgesia.

BACKGROUND: The orchiectomy in cats is a common surgical procedure with medium level of pain and for this reason requires intra and postoperative analgesia management. The aim of this study was to compare intra and postoperative pain in two groups of cats undergoing orchiectomy. Sixty healthy cats were randomly assigned in two groups (n = 30) to receive pre surgery ropivacaine hydrochloride (0.2 mL/kg at 0.5%) (R Group) or NaCl 0.9% (C group) into the spermatic cord. The intraoperative evaluation was carried out using the cardiorespiratory stability parameters and eventually administration of rescue analgesia. A rescue analgesia (fentanyl 2 µg/kg) was administered during orchiectomy in case of considerable increase of blood pressure, heart rate or respiratory rate. The postoperative evaluation was been done using scores following a UNESP-Botucatu multimodal scale for 6 h post-surgery. RESULTS: As result, cats in R group responded better to surgical procedure, maintaining lower postoperative pain scores than C group. CONCLUSIONS: The ultrasound-guided funicular block used in this study, as already demonstrated in dogs, is a good method to protect the cats from surgical pain and ensure a good level of surgical analgesia.

Analgesic comparison between topical irrigation (splash block) versus injection of lidocaine on the ovarian pedicle in canine ovariectomy.

OBJECTIVES: The aim of this study was to compare the analgesic efficacy of topical irrigation versus injection of lidocaine on the ovarian pedicle to provide analgesia in bitches ovariectomy. In the current study were monitored: increased blood pressure, heart rate and respiratory rate to identify an acute intraoperative nociceptive response. These parameters were registered at six times during the surgical procedure: grasping of the ovary (time 1), dissection of the mesosalpinx (time 2), tightening of the first loop ligature (time 3), tightening of the second loop ligature (time 4), transection of the ovarian pedicle (time 5) and release of the ovary (time 6). MATERIAL AND METHODS: Forty healthy bitches were randomly assigned in two groups (n = 20) to receive topical irrigation (splash block) of 2% lidocaine (C group) on both ovarian pedicle (2 mg/kg each), or an equal volume of lidocaine was injected at the same sites (R group). RESULTS: The results of the present study suggest that splash block may provide intraoperative analgesic effects equivalent to injection in the ovarian pedicle in dogs that have undergone ovariectomy. The lidocaine improved surgical analgesia during canine ovariectomy in both groups, and this action is not affected by the inoculation technique. CLINICAL SIGNIFICANCE: Pain management in veterinary patients is a crucial component of appropriate patient care. Therefore, the need for achieving safer anaesthesia for surgical intervention is gaining much attention. Ovariectomy is a common surgical procedure in bitches with medium level of pain. This study concluded that considering its relative simplicity, low cost, and safety, both techniques could be used in daily clinical practice.

Use of Contrast-Enhanced Ultrasound of the Testes after Non-Surgical Sterilization of Male Dogs with CaCl2 in Alcohol.

Sterilization by intratesticular injection of chemical agents is a non-surgical alternative to neutering male companion animals. We used contrast-enhanced ultrasound (CEUS) to monitor vascular alterations to testes immediately after the intratesticular injection of CaCL2 in alcohol. We evaluated the CEUS features of normal and damaged testes in 20 dogs after the intratesticular injection of CaCl2. The CEUS evaluation was performed at the site of the chemical agent inoculation. In treated testes, qualitative CEUS showed a lower intensity enhancement of the parenchyma than pre-treatment normal testes with a predominantly anechoic pattern and only a few hyperechoic vascular focal spots. Quantitative CEUS showed significantly lower values of time-intensity curve (TIC) parameters, including signal intensity (Peak: 4.72 ± 2.1), regional blood volume (RBV: 134.3 ± 63.7), and regional blood flow (RBF: 4.36 ± 2.18) than normal testes (p < 0.001). Sonographic findings from CEUS showed hypovascularization of the canine testicular parenchyma caused by the hardening agent. This diagnostic technique helps clinicians define testicular vascular alterations achieved by chemical castration more efficiently. Nevertheless, more studies are required to apply this methodology to more subjects with a broader weight range and stray dogs.

Effects of low-dose X-ray medical diagnostics on female gonads: Insights from large animal oocytes and human ovaries as complementary models.

Diagnostic imaging has significantly grown over the last thirty years as indispensable support for diagnostic, prognostic, therapeutic and monitoring procedures of human diseases. This study explored the effects of low-dose X-ray medical diagnostics exposure on female fertility. To aim this, cumulus-oocyte complexes (COCs) recovered from the ovaries of juvenile sheep and human ovaries were used as complementary models for in vitro studies. In the sheep model, the effects of low-dose X-rays on oocyte viability and developmental competence were evaluated. In human ovaries originated from two age group (21-25 and 33-36 years old) subjects with gender dysphoria, X-rays effects on tissue morphology, follicular density and expression of apoptosis-related (NOXA, PUMA, Bcl2, Bak, γH2AX) and cell cycle-related genes (p21 and ki67) were investigated. It was noted that in sheep, the minimum dose of 10 mGy did not influence most of examined parameters at oocyte and embryo levels, whereas 50 and 100 mGy X-ray exposure reduced oocyte bioenergetic/oxidative activity but without any visible effects on oocyte and embryo development. In addition, blastocyst bioenergetic/oxidative status was reduced with all used doses. Overall data on human ovaries showed that low-dose X-rays, similarly as in sheep, did not alter any of examined parameters. However, in women belonging to the 33-36 year group, significantly reduced follicular density was observed after exposure to 50 and 100 mGy, and increased NOXA and Bax expression after exposure at 50 mGy. In conclusion, used low-doses of X-ray exposure, which resemble doses used in medical diagnostics, produce weak damaging effects on female fertility with increased susceptibility in advanced age.

Effects of two different management systems on hormonal, behavioral, and semen quality in male dromedary camels.

Effects of two different management systems on male dromedary camel hormones, behaviors, and semen parameters were documented. Camels (n=6) were tested under two management systems: (i) housed in single boxes with 1-h freedom (H23); (ii) exposed to females for 17 h (from 3.30 p.m. to 8.30 a.m.) and then housed (ConExF). Blood was collected every morning; camel behavior was recorded twice a day: (i) from 7:00 to 8:00 a.m. to determine the short effects; (ii) from 2:00 to 3:00 p.m. to determine the long effects. Each camel underwent a female parade and semen collection thrice a week; sexual behavior, libido, and semen parameters were assessed. Testosterone and cortisol concentrations were higher in ConExF than H23. Compared to the H23 group, ConExF group spent more time walking, standing tripods, and looking outside their pen/box but they spent less time eating, ruminating, resting, standing, and showing stereotypical behaviors. In the morning, ConExF group spent more time walking, ruminating, and showing typical sexual behaviors compared to themselves during afternoon time and the H23 group. However, in the afternoon time, ConExF camels put more time their heads outside the box through the window and showed higher frequencies of stereotypies, probably due to a higher level of frustration. While the sexual behavioral score was higher and ejaculates showed a higher fraction of milky white and white-colored semen in ConExF than H23 group, their libido was similar. Overall, 17 h of exposure led to an increase in testosterone and cortisol levels, enhancing sexual behavior and semen color, but leading to frustration.

Seminal plasma Alters surface Glycoprofile of dromedary camel cryopreserved epididymal spermatozoa.

The high viscosity of Camelidae semen continues to present a major impediment for its application in assisted reproduction technology. The exposure of epididymal spermatozoa (ES) to seminal plasma (SP) may provide an approach to enhance the development of assisted reproductive techniques in these important domestic species. Since the sperm glycocalyx plays a key role in reproduction we aimed to evaluate whether SP exposure modifies the surface glycosylation patterns of cryopreserved dromedary ES. Epididymal sperm was collected through retrograde flushing of the cauda epididymidis that were obtained from orchidectomized mature dromedary bulls. The collected samples were then cryopreserved after dilution with a tris citrate clarified egg yolk extender, with and without the supplementation of 15% SP. Post-thaw carbohydrate surface profiles of both control and SP-treated spermatozoa were analyzed using 15 fluorescent lectins. Morpho-functional properties were also investigated via computer assisted sperm analysis. Lectin-binding analysis of the glycocalyx in control sperm revealed the presence of (1) N-glycans terminating with lactosamine (Con A, PHA-L, and RCA120), in both acrosomal and tail regions. Whilst (2) α2,3-/α2,6-linked sialic acids (MALII, SNA), and O-linked glycans terminating with a single N-acetylgalactosamine residue (Tn antigen) (HPA, SBA) along with galactoseß1,3N-acetylgalactosamine (T antigen) (PNA) were observed in the acrosomal cap. The expression of both N-acetylglucosamine (sWGA and GSA II) and terminalαgalactose (GSA I-B4) residues was also noted in the acrosomal cap region of control sperm. Compared with controls, SP treated samples displayed: 1) the appearance of bisected di-triantennary complex-type N-glycans (PHA-E), terminating with lactosamine, as well as an increase of O-glycans terminating with Tn and T antigens in both the acrosomal and tail regions; 2) an increase in glycans containing α2,6-linked sialic acid, N-acetylglucosamine, and αgalactose in the tail region. The cytoplasmic droplets of both control and seminal plasma-treated sperm bound Con A, PHA-E, PHA-L, RCA120, HPA, PNA, sWGA, GSA I-B4, and GSA II. These results indicate that SP treatment affects the glycan composition of the dromedary camel ES glycocalyx. More comprehensive studies are required in order to evaluate the fertilization capacity of SP-treated ES in order to facilitate its application in dromedary camel assisted reproduction technology.

Ochratoxin A affects oocyte maturation and subsequent embryo developmental dynamics in the juvenile sheep model.

The genotoxic and nephrotoxic mycotoxin Ochratoxin A (OTA) has also been reported to have adverse effects on oocyte maturation and embryo development. Previous studies on the effects of OTA on female fertility have used micromolar concentrations, but no information is available to date on effects in a more relevant nanomolar range. This study used a juvenile sheep model to evaluate the effects of oocyte exposure to low levels of OTA on maturation, fertilization, and embryo development. Further, it was investigated whether different mechanisms of action of OTA could be responsible for varying toxic effects at different levels of exposure. Cumulus-oocyte-complexes (COCs) were exposed to 10 µmol/L-0.1 nmol/L OTA during in vitro maturation and evaluated for cumulus viability, oocyte maturation, and bioenergetic/oxidative status. COCs were subjected to in vitro fertilization, embryo culture, and embryo quality assessment via morphology, viability, bioenergetic/oxidative status, and time-lapse monitoring. At micromolar concentrations, OTA induced cytotoxic effects, by reducing cumulus expansion and oocyte maturation. OTA altered temporospatial dynamics of zygote pronuclear formation and embryo morphokinetics. Blastocysts, even morphologically normal, were found to undergo collapse events, which were probably related to boosted blastocyst mitochondrial activity. At nanomolar concentrations, OTA did not affect COC morpho-functional parameters, but impaired oocyte ability to prevent polyspermy and increased blastocyst apoptosis. In conclusion, in the female germ cell, cytotoxic nonspecific effects characterize OTA-induced toxicity at high exposure levels, whereas fine tuning-mode effects, not associated with altered cell viability and integrity, characterize OTA toxic action at low levels.

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.

Retrograde flushing collection and freezing of dromedary camel epididymal spermatozoa with seminal plasma.

The objectives of this study were to describe the parameters of dromedary camel epididymal spermatozoa collected by retrograde flushing (RF) technique and to evaluate the freezability of the collected sperm, diluted with and without the supplementation of seminal plasma (SP). Two experiments were conducted: in Experiment 1, ES were recovered within 6-8 h after castration; selected samples were diluted with a Tris-citrate egg-yolk glycerolated buffer and frozen. In Experiment 2, epididymides were stored for 24 h at 4 °C before RF and semen samples were frozen after dilution with a Tris-lactose egg-yolk glycerolated extender with and without 15% SP. In Experiment 1, eight semen samples were obtained from ten epididymides with a mean of 500 × 106 total spermatozoa recovered, per flushed epididymis. Mean post-thaw motility and progressive motility were 75 and 17%, respectively. In Experiment 2, 15 samples were collected, out of the 18 epididymides (mean number of collected spermatozoa: 700 × 106), and 13 of these samples were of excellent quality. Post-thaw parameters were not satisfactory but the supplementation of the freezing medium with 15% SP improved the progressive motility and kinematic parameters of the spermatozoa.

Centrifugation Force and Time Alter CASA Parameters and Oxidative Status of Cryopreserved Stallion Sperm.

Conventional sperm selection techniques used in ARTs rely on centrifugation steps. To date, the different studies reported on the effects of centrifugation on stallion sperm motility provided contrasting results and do not include effects on mitochondrial functionality and different oxidative parameters. The effects of different centrifugation protocols (300 ×g for 5', 300 ×g for 10', 1500 ×g for 5' and 1500 ×g for 10' vs no centrifugation) on motility and oxidative status in cryopreserved stallion sperm, were analyzed. After centrifugation, almost all motility parameters were significantly altered, as observed by computer-assisted sperm analysis. A polarographic assay of oxygen consumption showed a progressive decrease in mitochondria respiration from the gentlest to the strongest protocol. By laser scanning confocal microscopy, significant reduction of mitochondrial membrane potential, at any tested protocol, and time-dependent effects, at the same centrifugal force, were found. Increased DNA fragmentation index at any tested protocol and time-dependent effects at the same centrifugal force were found, whereas increased protein carbonylation was observed only at the strongest centrifugal force. These results provide more comprehensive understandings on centrifugation-induced effects on cryopreserved stallion sperm and suggest that, even at a weak force for a short time, centrifugation impairs different aspects of equine sperm metabolism and functionality.

The mycotoxin beauvericin induces oocyte mitochondrial dysfunction and affects embryo development in the juvenile sheep.

Beauvericin (BEA) is a mycotoxin produced by Beauveria bassiana and Fusarium species recently reported as toxic on porcine oocyte maturation and embryo development. The aim of this study was to assess, in the juvenile sheep, whether its effects are due to alterations of oocyte and/or embryo bioenergetic/oxidative status. Cumulus-oocyte-complexes (COCs) were exposed to BEA during in vitro maturation (IVM), evaluated for cumulus cell (CC) apoptosis, oocyte maturation and bioenergetic/oxidative status or subjected to in vitro fertilization (IVF) and embryo culture (IVEC). Oocyte nuclear maturation and embryo development were assessed after Hoechst staining and CC apoptosis was analysed by terminal deoxynucleotidyl transferase-mediated dUTP nick-End labeling assay and chromatin morphology after Hoechst staining by epifluorescence microscopy. Oocyte and blastocyst bioenergetic/oxidative status were assessed by confocal microscopy after mitochondria and reactive oxygen species labelling with specific probes. BEA showed various toxic effects, that is, short-term effects on somatic and germinal compartment of the COC (CCs and the oocyte) and long-term carry-over effects on developing embryos. In detail, at 5 µM, it significantly reduced oocyte maturation and immature oocytes showed increased late-stage (Type C) CC apoptosis and DNA fragmentation while matured oocytes showed unaffected CC viability but abnormal mitochondrial distribution patterns. At lower tested concentrations (3-0.5 µM), BEA did not affect oocyte maturation, but matured oocytes showed reduced mitochondrial activity. At low concentrations, BEA impaired embryo developmental capacity and blastocyst quality after IVF and IVEC. In conclusion, in the juvenile sheep, COC exposure to BEA induces CC apoptosis and oocyte mitochondrial dysfunction with negative impact on embryo development.

Effects of intratesticular vs intraepididymal calcium chloride sterilant on testicular morphology and fertility in dogs.

BACKGROUND: Both stray and free-roaming owned dogs contribute to the serious global dog overpopulation problem. Many dog owners are unwilling to have their pet castrated for various reasons, including a reluctance to have their dog's behavior changed. A non-surgical method of sterilizing both stray and owned dogs would help to prevent unwanted litters. Previous studies have shown that intratesticular injection of calcium chloride dihydrate (CaCl2) in alcohol is a promising and cost-effective alternative to surgery for stray dogs, with testosterone significantly decreased and sexual activity eliminated. The aim of this study was to compare the use of a solution of 20% CaCl2 in 95% ethanol injected into the testicles or into the head of the epididymis. METHODS: A total of 148 dogs divided into 4 groups (2 experimental and 2 control) were respectively injected with CaCl2 or saline solution into the testicle or epididymal head (ultrasound-guided). The animals were examined at 0, 3, 6, and 9 months for sperm quality, concentration of testosterone in serum, and side effects; at 0 and 5 months with contrast-enhanced ultrasound (CEUS) to enhance the morphological aspects/alteration of the testicular parenchyma or epididymis; and at 9 months when all were castrated for histological examination. RESULTS: All dogs treated with CaCl2 became sterile with azoospermia achieved over the 9-month study. The concentration of testosterone in serum significantly decreased following intratesticular treatment with CaCl2. No adverse effects were noted. CONCLUSIONS: A single, bilateral intratesticular injection of 20% CaCl2 in 95% ethanol was confirmed to be a reliable method for induction of sterilization in male dogs. The approach showed long-term efficacy and may reduce sexual behavior, with the additional benefits of low-cost and ease of use, making this nonsurgical method appropriate for use in stray dogs. Sterility was also achieved if injected in the head of the epididymis but no significant decrease in serum concentration of testosterone occurred. Moreover, performing the intraepididymal injection into the epididymal head was as time consuming as orchiectomy. This approach may be optimal for use in owned dogs where anatomical integrity and testosterone maintenance is preferred by the owner.

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.