Does Placental Efficiency and Vascularization Affect Puppy Health? A Study in Boxer and Dobermann Dogs.

BACKGROUND: The anatomical and functional characteristics of the placenta influence the adaptive ability of the fetus to the extrauterine environment. Placental efficiency, measured as the gram of neonate produced by each gram of placenta, summarizes these characteristics. In the present study, placental efficiency and its impact on the 7-day postpartum life of the puppies were studied in canine large breeds. METHODS: Placental efficiency (PE) was computed using chorioallantois weight (WPE) and surface (SPE) efficiency for puppies born from natural delivery or elective cesarean section. Capillary density was also histologically determined. Neonate viability was estimated by the APGAR score and the daily weight gain (DWG) was recorded on day 7 after delivery. RESULTS: from 15 large-breed bitches, 69 live puppies were born by natural delivery (24 puppies) and elective cesarean section (45 puppies). Cluster analysis detected a group of neonates for which reduced placental efficiency (8 for the WPE, 9 for the SPE) was identified, despite a placental weight and surface within the mean and no difference in capillary density. In this group, the DWG was lower and the mortality within 7 days was higher. CONCLUSIONS: the results suggest that placental efficiency could be an additional tool for the evaluation of a puppy in the first 7 days after delivery.

Cushioned and high-speed centrifugation improve sperm recovery rate but affect the quality of fresh and cryopreserved feline spermatozoa.

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.

Ultrasound-Based Technologies for the Evaluation of Testicles in the Dog: Keystones and Breakthroughs.

Ultrasonography is a valuable diagnostic tool extensively used in the andrology of human and domestic animals, including dogs. This review aims to provide an overview of various technologies based on ultrasound, from the basic B-Mode ultrasonography to the more recent advancements, such as contrast-enhanced ultrasonography (CEUS) and ultrasound elastography (UEl), all of which are utilized in the evaluation of canine testicles. The review outlines the principles behind each of these technologies and discusses their application in assessing normal and abnormal testicular conditions. B-mode canine testicular ultrasonography primarily focuses on detecting focal lesions but has limitations in terms of objectivity. Other technologies, including Doppler ultrasonography, B-Flow, and CEUS, allow for the characterization of vascular patterns, which could be further measured using specific applications like spectral Doppler or quantitative CEUS. Additionally, ultrasound elastography enables the assessment of parenchyma stiffness both qualitatively and quantitatively. These ultrasound-based technologies play a crucial role in andrology by providing valuable information for evaluating testicular function and integrity, aiding in the identification of pathological conditions that may impact the health and quality of life of male dogs.

Validation of the volumetric flow cytometry for bovine sperm concentration.

Sperm concentration is a stronghold of the andrological evaluation and the production of insemination doses. The use of haemocytometers, although considered the gold standard, is difficult to apply in field conditions because it is subjective and time-consuming. The present study was designed to validate the volumetric flow cytometry (volFC) in order to estimate bovine sperm concentration, comparing it with the performances of haemocytometer, NucleoCounter, and flow cytometry with the use of fluorospheres. Compared with other methods, volFC appeared less affected by large dilution of the sample, with similar concentrations calculated in the range of dilution 1:200-1:800. Using volFc the population detected on the basis of morphological criteria and fluorescence of DNA better represents the real concentration of sperm in the sample. The volFC showed high repeatability compared with the haemocytometer (coefficient of variation 1.85% and 4.52%, respectively) and stable performances with cryopreserved samples, with negligible effects of the medium components. The present study showed that volFC is as accurate and precise as other techniques to estimate sperm concentration in bovine fresh and frozen semen, but it is less affected by operative conditions, such as sample dilution. The possibility to quantify sperm functional subpopulations by volFC could potentially implement the study of the relationship between sperm attributes and fertility.

Stiffness estimated by strain elastography reflects canine testicular spermatogenesis and histology.

Ultrasound elastography was proposed for the evaluation of testicular focal lesions, but no studies verified the agreement between the whole histological architecture of the testis and the stiffness measured by elastography. The present study explored the use of strain elastography in the evaluation of testis with normal or abnormal spermatogenesis, classified based on epididymal sperm attributes, and the consistency between elastographic parameters and the testicular histological feature. Strain elastography was performed during the routine andrological examination in 22 dogs presented for elective orchiectomy. Epididymal sperm attributes and testicular histology were analyzed after orchiectomy. Based on the epididymal sperm characteristics, testes were classified according to normal or abnormal spermatogenesis, and strain elastographic attributes were compared between groups. Possible correlations between strain elastography and histological features were also explored. Consistent with the literature in humans, testes with abnormal spermatogenesis were stiffer (mean strain elastographic index 3.6 ± 0.6) compared with normal testes (mean strain elastographic index 1.9 ± 0.2; P < 0.01). The strain elastographic index was negatively correlated with the area occupied by seminiferous tubules (Pearson's rho = -0.716; P = 0.0003), the mean diameter (Pearson's rho = -0.742; P = 0.0002), and thickness of the seminiferous tubule (Pearson's rho = -0.728; P = 0.0002). Surprisingly, no correlations were found between the area occupied by connective tissue in histological sections and elastographic attributes, suggesting that the increased stiffness was not related to the increased amount of connective tissue. This study demonstrated that strain elastography could be used to support the andrological examination, but measurements should be acquired in specific regions to be reliable.

Strain and Shear-Wave Elastography and Their Relationship to Histopathological Features of Canine Mammary Nodular Lesions.

Mammary gland tumours have a significant impact on the health of dogs, requiring diagnostic tools to support clinicians to develop appropriate therapeutic strategies. Sonoelastography is an emerging technology that is able to define the stiffness of the tissue and has promising applications in the evaluation of mammary gland lesions. In the present study, strain elastography (STE) and shear-wave (SWE) elastography were compared in 38 mammary nodular lesions for their ability to define the histopathological features of canine mammary lesions. Among the techniques, SWE showed better repeatability (intraclass correlation coefficient: 0.876), whereas STE was found to be only acceptable (intraclass correlation coefficient: 0.456). Mammary nodular lesions showed a wide range of tissue stiffening with a similar mean value for STE and SWE in benign (4 ± 0.3 and 115.4 ± 12.6 kPa, respectively) and malignant lesions (3.8 ± 0.1 and 115.5 ± 4.5 kPa, respectively). A significant correlation was found between lesion fibrosis and STE (STE-I: r = 0.513, p < 0.001; STE-R: r = 0.591, p < 0.001) or SWE-S (r = 0.769; p < 0.001). In conclusion, SWE was reliable and correlated with fibrosis and was similar for both benign and malignant lesions, suggesting that other collateral diagnostic techniques should be considered in conjunction with SWE to characterize mammary nodular lesions in dogs.

Evaluation of Brix Refractometry to Estimate Immunoglobulin G Content in Buffalo Colostrum and Neonatal Calf Serum.

Brix refractometry has been widely demonstrated to be a useful tool for monitoring colostrum management program and passive immunity transfer (PIT) in Bovines, but its suitability has never been verified in Buffalo. Therefore, the objective of this study was to evaluate the utility of a simple and rapid tool such as a digital Brix refractometer to estimate colostrum quality and for evaluating the success of passive transfer of immunoglobulin G (IgG) in Buffalo calves. The optimal cut points levels for Brix Refractometry for distinguishing good- and poor-quality colostrum and for assessing the adequacy of passive immunity transfer in calves were determined. For this aim, 26 first-milking maternal colostrum (MC) were collected from first-calf heifers. Blood samples were obtained from their calves at birth (T0) and 72 hours after (T3). Colostrum and Serum IgG content were determined by indirect enzyme-linked immunosorbent assay (ELISA), whereas total protein (TP, g/dL) and percentage Brix (%Brix) by means of a digital Brix refractometer. The mean colostrum IgG was 64.9 ± 29.3 mg/mL. The mean serum %Brix at T3 was 9.6 ± 0.9 %. The mean serum IgG content at T3 was 11.1 ± 2.0 mg/mL. Pearson's correlation coefficient (rp) was determined between Brix and ELISA measurements: colostrum %Brix showed a significant correlation with serum %Brix (rp = 0.82, p < 0.001); serum %Brix was highly correlated with serum TP (STP, g/dL) (rp = 0.98, p < 0.001) and serum IgG (mg/mL) (rp = 0.85, p < 0.001). A cut point of 18% Brix to estimate samples of MC ≥ 50 mg/mL from first-calf heifers was more appropriate for the buffalo. A cut point of 8.4% Brix resulted in the greatest percentage of calf serum samples being correctly classified. Based on our findings, a digital Brix refractometer could be a useful tool to monitor colostrum quality and to estimate PIT in Buffalo calves.

Kisspeptin Receptor on the Sperm Surface Reflects Epididymal Maturation in the Dog.

Alongside the well-known central modulatory role, the Kisspeptin system, comprising Kiss1, its cleavage products (Kisspeptins), and Kisspeptin receptor (Kiss1R), was found to regulate gonadal functions in vertebrates; however, its functional role in the male gamete and its localization during maturation have been poorly understood. The present study analyzed Kisspeptin system in dog testis and spermatozoa recovered from different segments of the epididymis, with focus on Kiss1R on sperm surface alongside the maturation during epididymal transit, demonstrated by modification in sperm kinetic, morphology, and protamination. The proteins Kiss1 and Kiss1R were detected in dog testis. The receptor Kiss1R only was detected in total protein extracts from epididymis spermatozoa, whereas dot blot revealed Kiss1 immunoreactivity in the epidydimal fluid. An increase of the Kiss1R protein on sperm surface along the length of the epididymis, with spermatozoa in the tail showing plasma membrane integrity and Kiss1R protein (p < 0.05 vs. epididymis head and body) was observed by flow cytometry and further confirmed by epifluorescence microscopy and Western blot carried on sperm membrane preparations. In parallel, during the transit in the epididymis spermatozoa significantly modified their ability to move and the pattern of motility; a progressive increase in protaminization also occurred. In conclusion, Kisspeptin system was detected in dog testis and spermatozoa. Kiss1R trafficking toward plasma membrane along the length of the epididymis and Kiss1 in epididymal fluid suggested a new functional role of the Kisspeptin system in sperm maturation and storage.

Environmental temperature and relative humidity differently affect the sperm characteristics in Brown Swiss and Belgian Blue bulls.

In a temperature-increasing scenario, due to global warming, the individual thermic resilience of the male assumes a crucial role in the reproductive efficiency of a male since the thermic stress, such as the inability of the male to reduce body or regional temperature on a physiological level, impairs testicular function. In this study, the effect of the environmental conditions on the fresh semen quality, in terms of volume, concentration, total sperm in the ejaculate, total motility, normal morphology, membrane integrity, and discarding rate, were compared longitudinally in Belgian Blue (BB) and Brown Swiss (BS) bulls. The environmental conditions, summarized in the mean temperature-humidity index (THI), were calculated on the day of collection, as well as 7 days (epididymal maturation), 35 days (late spermatogenesis), and 70 days (early spermatogenesis) before the collection, to reflect spermatogenesis time. Our findings showed that limited seasonal effects were present in the semen quality of BS bulls. On the other hand, in BB bulls lower semen quality was found between July and November, with a different timing depending on the seminal parameter. This effect of the season on BB semen parameters appears to be related to the THI. The data presented in this study shows that the temperature and humidity, summarized in THI, could affect the semen quality of the bull on breed basis, given that volume, concentration, total sperm in the ejaculate, total motility, membrane integrity, and sperm normal morphology were significantly reduced by an increasing THI in the Belgian Blue bulls, but not in Brown Swiss bulls.

Maternal and Neonatal Behaviour in Italian Mediterranean Buffaloes.

The aim of this study was to describe the neonatal and maternal behaviour of Italian Mediterranean buffaloes. Thirty primiparous buffaloes were moved into individual pens 12.5 (±2.5) days before calving. Maternal and neonatal behaviours were recorded for 48 h after calving and the analysis was performed in continuous sampling with the software BORIS. Calves' clinical evaluations (temperature, weight, and heart and respiratory rates) were performed at different time intervals and correlated with behavioural data from the dam. Data were analysed with parametric and non-parametric methods after controlling their distribution. The maternal behavioural pattern found highlighted buffaloes' priorities during the post-partum period: firstly, they stand and start grooming to ensure proper care for the calf; it is only after this that they dedicate time to maintenance behaviours (feeding and lying). The dams mainly groomed the calf during the first six hours after calving (average time in the 1-6-h interval: 7.7 ± 2.5 min., F = (2.5, 60.2) = 75.0; p < 0.001) to ensure the formation of the mother-infant bond; thereafter, the behaviour decreased over time. As reported in the literature, inexperienced mothers could sometimes delay the calf's first suckling with aggressive or rejection behaviours. In this regard, 16 buffalo dams showed at least one maternal rejection behaviour, which was found to negatively correlate with calves' daily weight gain (DWG) at 14 (rs = -0.5, p = 0.02) and 21 days (rs = -0.7, p < 0.001). The calves took on average 212.0 ± 110.0 min to suckle, and this behaviour was mainly shown during the first six hours. Overall, suckling behaviour was correlated with standing: (rs = 0.6, p < 0.001) and walking (rs = 0.9, p < 0.001). The calves' live weight and DWG were consistently higher than the values reported in the literature. Our results present a detailed description of maternal and neonatal behaviour in the early post-partum period in Italian Mediterranean buffaloes. We also found that maternal rejection behaviours can negatively influence the calves' growth. Finally, we think that such results can improve the management of buffaloes during the period around parturition.

Osmotic tolerance of rabbit spermatozoa is affected by extender composition and temperature.

Sperm osmotic adaptability to anisosmotic conditions is important for sperm epididymal maturation, motility activation at ejaculation, and female tract colonization, or for conducting technological procedures such as cryopreservation. Several factors affect this adaptability, including the fluid composition that contributes to water flow dynamics, and the temperature at which osmotic stress is initiated. This study was designed to investigate the effect of medium composition (electrolyte- or sugar-based extender) and temperature (25 and 5 °C) on rabbit sperm adaptability to anisosmotic conditions. Rabbit spermatozoa, therefore, were diluted at both temperatures (25 and 5 °C) in electrolyte- or sugar-based media at increasing osmotic conditions (100 to 1,000 mOsm/kg), and values for sperm variables (sperm kinetics, membrane integrity, mitochondrial membrane potential) were estimated as endpoints. Sperm kinetics seemed to be more sensitive to osmotic stress than membrane integrity or mitochondrial function. The effect of moderate hypoosmotic stress did not differ when there was use of sugar- and electrolyte-based extenders at 25 °C (P > 0.05). In hyper-tonic conditions at 25 °C, the sugar-based extender was more effective in protecting sperm membrane integrity and mitochondrial function (P < 0.05). The lesser temperature made the differences more relevant because of the detrimental effect of hyperosmotic stress was more evident in the electrolyte-based extender at 5 °C (P < 0.05). The results from this study indicated rabbit spermatozoa have different adaptability to anisosmotic conditions induced by sugar- and electrolyte-based media and that the temperature at which the osmotic stress is initiated affects the cellular response.

Pulse-wave Doppler pulsatility and resistive indexes of the testicular artery increase in canine testis with abnormal spermatogenesis.

Pulse-wave Doppler (PWD) ultrasonography of the testicular artery was recently used to compare parameters in fertile and infertile males, or males with normal and abnormal semen quality, while no studies verified the concomitant histological feature of the testis. The experimental hypothesis of the present study was that the blood flow parameters measured by PWD at different levels of the testicular artery reflect sperm attributes evaluated on spermatozoa collected from the tail of the epididymis and testicular histological characteristics. Pulse-wave Doppler parameters were recorded on both testes from 16 dogs of unknown fertility. After routine castration, epididymal spermatozoa were collected and evaluated for sperm total and progressive motility, membrane integrity, and sperm morphology. Testes were submitted to histological and immunohistochemical evaluations to quantify microscopic attributes and connective tissue and vascular area extent, respectively. The findings showed that the increased PWD indexes were suggestive of testicular histological alteration since negative correlations were found between pulsatility index and resistive index, measured in the marginal testicular artery, and histological attributes such as the seminiferous epithelial thickness (R = -0.528, P = 0.016 and R = -0.554, P = 0.005, respectively) and the tubular area/total area ratio (R = -0.494, P = 0.024 and R = -0.563, P = 0.004, respectively). Some histological characteristics, such as tubular diameter, seminiferous epithelium thickness, and the tubular area/total area ratio of the testis are correlated with the percentage of primary abnormalities (R = -0.874, P = 0.001, and R = -0.847, P = 0.001, and R = - 0.865, P = 0.001, respectively). In testes with increased sperm primary abnormalities, higher pulsatility index was found compared with testes with normal spermatogenesis in supratesticular artery (1.07 ± 0.06 vs 0.82 ± 0.23, P < 0.05), marginal artery (0.73 ± 0.04 vs 0.61 ± 0.07, P < 0.05), and intratesticular artery (0.67 ± 0.03 vs 0.43 ± 0.1, P < 0.05). Similarly, the resistive index was found significantly higher in the same vessel tracts in testes with abnormal and normal spermatogenesis (0.62 ± 0.02, 0.53 ± 0.02, and 0.46 ± 0.02 vs 0.51 ± 0.06, 0.44 ± 0.04, and 0.32 ± 0.06, respectively; P < 0.05). Our data demonstrated for the first time the relationship between PWD indexes and the contextual testicular architecture, suggesting that the pulsatility and resistive indexes could be an effective tool in andrology. In light of different histological alterations in the testis, caution is warranted in the definition of thresholds for normal and abnormal testicular blood flow indexes.

Venous blood gas parameters in healthy Mediterranean buffalo calves in the first 72 hours of life.

In the first hours after calving the neonate radically modifies its anatomy and physiology to adapt to extrauterine life. These modifications, however, make the calf prone to respiratory and metabolic dysfunction. Thus, stillbirth, defined as the death of the neonate within 48 h after birth, represents a relevant issue for buffalo farming. Of the clinical procedures, the blood gas analysis of venous samples provides useful information regarding the acid-base status of the neonate, allowing the optimal management of potential illness calves. However, the blood gas parameters have never been studied in the buffalo calf in the first 72 h of life. Thus, this study aimed to report blood gas parameters at calving and 1, 12, 24, and 72 h after birth in healthy buffalo neonates born after normal parturition. The study involved 20 healthy buffalo calves, born from normal pregnancy and parturition. The parturition and the first hours after were monitored by closed-circuit television cameras, and within 1 min after calving the buffalo neonate received a clinical evaluation (APGAR, temperature), venous blood sampling for blood gas (pH, partial pressure carbon dioxide - pCO2, partial pressure oxygen - pO2, base excess, bicarbonate concentration, oxygen saturation, total carbon dioxide - tCO2, and lactate), and glycemia analysis. The same evaluations were performed after 1, 12, 24, and 72 h. The present study showed a moderate mixed acidosis at calving, similar to that reported in the bovine neonate, likely due to the high venous blood pCO2 and lactate. This acidemia resolved spontaneously within 12 h as a reduction of pCO2 (P ≤ 0.05). Lactate reduced significantly (P ≤ 0.05) only at 72 h, similarly to the trend reported previously for the bovine calf. Blood lactate concentration was correlated with glycemia at calving and 1 h (r = 0.677, P = 0.014), and with stage 2 duration (r = 0.647, P = 0.006). These findings support a possible mechanism of glucose generation induced by catecholamines coupled to anaerobic metabolism soon after calving. After calving, all the blood gas parameters changed rapidly reaching values comparable with those reported in the literature for older buffalo calves from 72 h after calving. No correlations were found between APGAR score and venous blood gas parameters, likely due to the inclusion criteria that selected only health buffalo calves born after normal parturition. The findings suggested that the venous blood gas analysis could be considered supportive rather than an alternative to the clinical evaluation of the buffalo calf.

Is the protective effect of egg yolk against osmotic and cryogenic damage on dog spermatozoa dose-dependent?

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 %).

Abundances of autophagy-related protein LC3B in granulosa cells, cumulus cells, and oocytes during atresia of pig antral follicles.

In mammals, apoptosis has been accepted as the type of programmed cell death (PCD) that occurs in ovarian follicles undergoing atresia. Results of recent studies, however, indicate autophagy may be an alternative mechanism involved in follicle depletion through independent or tandem actions with apoptosis. Western blotting and immunofluorescence procedures were used in the present study to investigate the abundances of LC3B protein in freshly collected granulosa cells (GCs), cumulus cells (CCs), and oocytes to evaluate whether autophagy is an important process of antral follicle atresia in sexually mature sows. Furthermore, apoptosis was analyzed using annexin V and TUNEL assays in the same cellular cohorts to evaluate the correlation between the two processes. Immunostaining results indicate autophagy was induced in the majority of GCs, CCs, and oocytes from early and advanced stage atretic follicles. The quantitative results of western blot analysis indicate there is a progressive increase (P <  0.05) in abundance of autophagy-related protein (LC3B-II) in these cells compared with cells in non-atretic follicles. Furthermore, there is confirmation that apoptosis occurs in the GCs of atretic follicles, thus indicating that in pigs apoptosis and autophagy are processes in GCs that regulate PCD and as a consequence antral follicle depletion. There was a greater abundance of LC3B-II in CCs and oocytes of atretic follicles, while apoptosis was not detected. It, therefore, is suggested that in these cells the two processes function independently, with autophagy having a cytoprotective rather than PCD mechanism of action.

Cryopreservation of turkey spermatozoa without permeant cryoprotectants.

In avian species, cryopreservation of semen is necessary for developing sperm cryobanks. It is very difficult, however to cryopreserve turkey sperm and have sperm be viable after thawing. Glycerol, the commonly used sperm cryoprotectant in many species, is toxic to sperm of avian species. The aim of this study was to evaluate whether the non-permeating dextran was effective for the cryopreservation and maintenance of turkey spermatozoa viability after thawing, avoiding the use of permeating cryoprotectants. Turkey sperm were diluted with a medium supplemented with 11% glycerol or dextran with a 1,000 molecular weight (MW), dextran with a 10,000 MW, or dextran with a 20,000 MW each at a 2%, 5%, or 10% concentration. Sperm kinetic characteristics, membrane and acrosome integrity (AI), and the capacity of spermatozoa to interact with the autologous perivitelline layer were evaluated after equilibration and cryopreservation. Results indicate that with use of glycerol and the 1,000 MW dextran there was lesser sperm viability after both equilibration and cryopreservation, compared with use of the 10,000 or 20,000 MW dextran compounds. There was a greater cryoprotective effect with the 10,000 and 20,000 MW dextran compounds at the 10% concentration with spermatozoa maintaining a greater functionality and capacity to interact with the autologous perivitelline layer. In conclusion, the results of this study indicate turkey spermatozoa could be effectively cryopreserved in extender without the use of glycerol as a penetrating cryoprotectant but with the use of the 10,000 or 20,000 MW dextran compounds at a 10% extender concentration.

Congenital lung displacement in an English bulldog puppy.

The English bulldog is notoriously predisposed to a number of disease conditions, including congenital defects, likely due to extreme genetic selection which has reduced genetic variability within this breed. The present report describes, for the first time, the main clinical and pathological features of congenital lung displacement in an English bulldog puppy. The defect involved the entire left lung, which was normally developed but rotated and shifted to the right hemithorax. The pathogenesis of congenital anomalies is largely unknown. characterisation of the genetic bases of these malformations is crucial for a reform of the breeding selection which is also aimed at correcting phenotypic abnormalities with a strong impact on individual health.

Effect of dietary grape marc on fresh and refrigerated boar semen.

In several studies there has been evaluation of the dietary addition of antioxidants to improve the quality of fresh and stored semen in domestic animals. Grape marc (GM), as the residue of vinification, contains large amounts of polyphenols with antioxidant, anti-inflammatory, antimicrobial, and antiaging effects. In this study, two regimens of dietary GM supplementation (2% and 4%) were tested regarding effects on the characteristics and lipid peroxidation of fresh and stored semen. The dietary supplementation of GM improved sperm characteristics in fresh semen, especially at 4% of GM. There were greater values for all kinetic variables and membrane integrity, and lesser values for sperm abnormalities and lipid peroxidation with 2% and 4% GM supplementation. Lipid peroxidation of the pellet was less with both 2% and 4% GM supplementation. During sperm storage, the dietary supplementation of GM improved the quality of sperm, with greater values for kinetic variables and membrane integrity at day 15 of storage. In the present study, supplementation of GM in boar diets improved fresh semen characteristics and reduced the lipid peroxidation of ejaculated spermatozoa, possibly due to the effect of polyphenols present in the GM. Consequently, this likely resulted in improved sperm quality during storage.

Biochemical blood analysis along pregnancy in Martina Franca jennies.

The availability of biochemical blood data specific for the pregnant condition are essential for the correct management of both normal pregnancies and also for the prompt recognition of every abnormality. Because of the lacking knowledge about biochemical blood analysis in the donkey along the entire pregnancy, the study was designed to provide first preliminary data about the values and possible changes of blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), alkaline phosphatase (ALP), creatine-kinase (CK), blood urea nitrogen (BUN), creatinine (CREA), uric acid (UA), amylase (AMY), gamma-glutamyl transferase (γ-GT), triglycerides (TRI), cholesterol (CHOL), total protein (TP), albumin (ALB), glucose (GLU), phosphorus (P), calcium (Ca), occurring from the beginning of pregnancy until parturition. The study was performed on 10 Martina Franca healthy jennies with normal pregnancy course and giving birth to mature, healthy and viable foals. Blood samples were collected monthly from the 1st to the 6th month of pregnancy, then twice a month from the 6th to the 9th month and afterwards weekly until parturition. The results showed a significant slight increase of glucose and creatinine in the second quarter of pregnancy and a minor decrease of cholesterol near to parturition, while all the other parameters did not significantly change along pregnancy.