This review provides a comprehensive exploration of the applications and advancements in ultrasonographic techniques for evaluating the uterus in domestic canines. Conventional grey-scale ultrasonography remains a cornerstone for detecting anatomical and pathological changes, while emerging modalities such as Doppler ultrasonography, contrast-enhanced ultrasonography (CEUS), and ultrasound elastography (UEl) have expanded diagnostic capabilities by providing insights into vascularity, tissue stiffness, and microvascular perfusion. Recent innovations in three-dimensional (3D) and four-dimensional (4D) ultrasonography have further revolutionized imaging by enabling detailed visualization of fetal anatomy and dynamic intrauterine processes. While these techniques have been extensively studied in human and veterinary medicine for various organs, their application to the canine uterus, both non-pregnant and pregnant, remains underexplored. This review bridges gaps in existing literature by incorporating findings from related fields and outlining future directions for research to improve the diagnosis and management of reproductive uterine disorders in dogs.

Plastic contamination (plastamination) is one of the main challenges of the 21st century. Microplastics (MPs, 5 mm-1 µm) and nanoplastics (NPs, <1 µm) are mainly produced from the environmental degradation of plastic waste and enter the food chain, bypass biological barriers, and bioaccumulate in tissues where they exert toxic/inflammatory effects. In the male reproductive system, MP/NPs can cross the blood-testis barrier, impair spermatogenesis, and affect semen quality parameters. While biodegradable polymers like polylactic acid (PLA) may represent an ecofriendly alternative to carbon fossil polymers, their real effects in reproduction have been poorly investigated. In the present manuscript, the effects of increasing doses of rhodamine B-loaded PLA-NPs (170 ± 20 nm mean size) on the physiology of frozen-thawed bovine sperm were investigated using a computer-assisted sperm analyzer (CASA) and flow cytometry. Sperm kinetics revealed a significant effect of PLA-NPs on progressive motility at 60 min, with higher values at 200 and 300 µg/mL doses (p < 0.05 vs. control). Flow cytometry showed that PLA-NP exposure did not alter acrosomal integrity (PNA488+); however, the proportion of spermatozoa showing high mitochondrial potential (MitoDR+) was significantly reduced compared to controls at a 300 µg/mL PLA-NP dose after 120 min of incubation. The proportion of spermatozoa metabolically active (MitoDR+) with contextual membrane destabilization (M540+) was significantly lower in samples with 200 and 300 µg/mL PLA, independently of the incubation time. Finally, immunofluorescence analysis revealed the internalization of PLA-NPs within spermatozoa at the longest exposure time. In conclusion, PLA-NPs are internalized in spermatozoa, with specific localization in mitochondria, and slightly affect progressive motility mainly by mitochondrial interference and plasma membrane destabilization. Despite limited effects on post-thaw bull sperm motility even at high doses, plastamination warrants consideration, and further studies on the biological effects of biodegradable plastics are recommended to preserve sperm physiology.

The cryopreservation of epididymal spermatozoa is essential for conserving genetic material from endangered species of high-genetic value males suddenly die. The most effective extenders for cryopreservation contain glycerol and egg yolk as cryoprotective components, but variability in composition, contamination risks, and potential toxicity, make the need to find more effective and safe alternatives crucial. This study evaluated the effectiveness of dextran as a substitute for egg yolk and glycerol in feline epididymal sperm cryopreservation. Spermatozoa collected from 24 domestic tomcats after routine orchiectomy were pooled in 8 samples. Seven TRIS-based extenders were tested on these pools, containing dextran (10,000 or 40,000 MW) with varying glycerol concentrations (0 %, 2.5 %, or 5 %), compared to a conventional TRIS extender (20 % egg yolk, 5 % glycerol). Pre- and post-cryopreservation assessments included sperm kinetics (CASA), flow cytometric sperm integrity and function, and hyaluronan-binding ability. Dextran 10,000 MW extenders resulted in similar (in glycerol-free formulation) or improved (with glycerol) sperm quality compared to the control, with higher post-thaw total and progressive motility and cell function (P < 0.05). Dextran 40,000 MW did not result in a similar improvement. Dextran 10,000 MW is a promising alternative to egg yolk in feline sperm cryopreservation, reducing reliance on animal-derived components and glycerol. These findings support the development of safer, more standardised cryopreservation protocols, with potential applications in endangered felid conservation. Further studies are needed to refine dextran-based extenders for broader use.

This study evaluated B-mode ultrasonography and ultrasound elastography (UEl) for detecting structural and consistency changes in the canine cervix across the oestrous cycle, as the use of ultrasonographic techniques for this purpose remains underexplored. The present study aims to assess cervical features via ultrasound, acknowledging the critical role of the cervix in the canine oestrus cycle. Thirty-five bitches were evaluated during pro-oestrus, pre-ovulatory oestrus, peri-ovulatory oestrus, dioestrus and anoestrus. Progesterone levels, clinical signs, vaginoscopy and cytology were used to define the oestrous cycle phase. Cervical length, diameter, and the elastographic index (ElI) were measured, and the elastographic ratio (ElR) was calculated to compare cervical stiffness to the surrounding tissue. Cervical length and diameter values were observed to be higher during the pre-ovulatory and peri-ovulatory phases of oestrus (p < 0.05). ElI values in anoestrus were similar to dioestrus and significantly higher than pro-oestrus and pre-ovulatory oestrus (p < 0.05), reflecting greater cervical stiffness in anoestrus and dioestrus, while pro-oestrus and pre-ovulatory phases showed softer tissues. These findings underscore the utility of elastography in quantifying cervical tissue consistency and its correlation with hormonal influences, providing a novel diagnostic perspective for a more comprehensive understanding of reproductive health.

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.

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.

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.

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.

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 & PLUSMN; 0.6) compared with normal testes (mean strain elastographic index 1.9 & PLUSMN; 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. & COPY; 2023 Elsevier Inc. All rights reserved.

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.

In humans and animals there is a popular belief that spontaneous parturition, and therefore pregnancy length, could be influenced by the lunar cycle, but results from humans are often impaired by multiple variables of the studied population. In domestic animals those problems can be limited by studying restricted population, such as a single breed, reared under uniform conditions. The present study aimed to assess the possible influence of the lunar cycle on pregnancy length in Martina Franca jennies belonging to a single breeding farm. The retrospective study was performed on data collected from 96 Martina Franca healthy jennies with a normal singleton pregnancy course. The present study showed that, whit lunar cycle divided in 4 lunar phases, neither lunar phase at ovulation, nor lunar phase at foaling significantly influenced pregnancy length (371.8±6.45 days). Pregnancy length was not significantly influenced also by the month of the year in which ovulation and spontaneous foaling occurred, as well as by jennies’ age and foal’s sex. In conclusion, pregnancy length in Martina Franca jennies is very variable (335-395 days) but does not seem to be influenced by lunar phases at ovulation or at foaling, month of ovulation and foaling, and by jennies’ age and foal’s sex. Further studies are needed to clarify the reasons for this pregnancy length variability in Martina Franca donkey breed.

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.

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.

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.

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.

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.

Mules have been enrolled for agricultural work in national parks or islands and hard-to-reach places in which the use of machinery is forbidden or not feasible. Despite the importance of this animal, limited information is reported regarding gestation, parturition, and post-natal growth in mule. To evaluate the effects of the pregnancy length, birthweight, and fetal sex on neonatal health and growth in mule and donkey foals born from Heavy Draft mares (n=15) and Martina Franca jennies (n=18) inseminated with the same jackass until 30 months of age. This study was conducted on 15 healthy Heavy Draft mares, 4 to 6 years old and weighing 640 to 730 kg, and 18 healthy Martina Franca jennies 4 to 6 years old and weighing 390 to 420 kg. All animals presented normal reproductive history such as cyclicity, pregnancy, and foaling. The semen used for artificial insemination was collected from the same 7 years old 485 kg bodyweight Martina Franca jackass of proven fertility. Pregnancy length in the mares bearing mule foal (339 +/- 9 days) was similar to that reported in the equine gestation and was longer in female mules (347 +/- 4.4 days) than males (331 +/- 5.46 days) (P < 0.05). In donkey, pregnancy length was significantly longer in male foals (373.7 +/- 10.4 days) than females (366.9 +/- 5.6 days) (P < 0.01). The mean birth weight was higher in mules compared to donkey foals (50.6 +/- 4.83 vs 33.4 +/- 5.7 kg) and was greater in male mules than in females (53.95 +/- 3.2 kg and 46.6 +/- 2.96 kg, respectively; P < 0.05). Mule foal's weight increased faster than donkey foals, despite a similar height at withers at different time points. Pregnancy length affected birthweight in both mule and donkey foals, and birthweight was related to postnatal growth in both mule and donkey foals. The findings reported in this manuscript could optimize the maternal-foal management of this equine hybrid.

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.

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. (C) 2020 Published by Elsevier Inc.