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Different methods to measure tissue perfusion are available in equine neonatology, but they are not representative of microvascular derangements.To evaluate the feasibility of the sidestream dark-field (SDF) capillaroscopy to visualize the capillary microvasculature in conscious newborn foals, the differences between two imaging sites and times of measurements, and the measurements' reproducibility.Seventeen healthy newborn foals were enrolled. Three sites at the upper and lower lip mucosa were assessed by SDF, using a hand-held capiscope, at 24 h and at 4-5 days after birth. Videos were assessed independently by two observers for quality and for semiquantitative calculation of microvascular parameters, including vascular density (VD), microvascular flow index (MFI), proportion of perfused vessels (PPV), and functional capillary density (FCD). Data were analyzed using unpaired and paired Student's T-tests to assess differences between sites and time-points. Bland-Altman plots and intraclass correlation coefficient (ICC) were used to assess measurement reproducibility.Differences were found between the upper and the lower lip for VD at both 24 h and 4-5 days, and for FCD at 24 h, and between the two time-points for PPV at the lower lip. ICC for measurement reproducibility was good for all parameters (0.64-0.79) for the lower lip, and was good for VD and FCD (0,76-0,79) and fair to moderate for MFI and PPV (0.31-0.41) for the upper lip.Measurement of the capillary microvasculature is feasible in the conscious newborn foal. The lower lip has the best measurement reproducibility. Further investigations are warranted in cardiovascularly compromised cases, particularly in septic foals.

This study aimed to determine alpha-fetoprotein (AFP) concentrations in amniotic fluid, plasma of mares and respective foals: carrying normal pregnancies and delivering healthy foals (n=20; Group 1); carrying apparently normal pregnancies and delivering sick foals (n=15; Group 2); carrying high-risk pregnancies and delivering sick foals (n=14; Group 3). High-risk pregnancy was defined by a history of premature udder development/lactation or increased of the combined thickness of the uterus and placenta, or vulvar discharge and/or mares' systemic illness. Sick foals were affected by neonatal encephalopathy, sepsis, prematurity/dysmaturity, or hypoxic-ischemic encephalopathy. Based on histological examination of the chorioallantois, AFP trend was analyzed in pregnancies with pathologic (PFM) and normal fetal membranes (NFM). Concentrations of AFP were measured using a commercially available immunoassay previously validated for horses. Mares' plasma AFP did not change during the last 15-20 days of pregnancy in the three groups, and there was no difference among them. Amniotic fluid AFP was higher in Group 3 (P=.014). Foals' plasma AFP concentration was higher from birth to 72hours in foals of Group 2 and 3 than in healthy ones, and foals of Group 3 had the highest value. The strong association (r=0.84; P < .0001) between AFP in amniotic fluid and foals' plasma at birth is likely due to the presence of AFP in fetal urine. AFP was higher in pregnancy with PFM than with NFM in mare's plasma at admission (P=.031), amniotic fluid (P=.004), foal's plasma at birth (P=.002), at 24 (P=.005) and at 72 hours of life (P=.004). AFP is higher in pregnancy with histopathological lesions of the chorioallantois providing the evidence of the differences between pregnancy with a normal placental barrier and the more compromised ones. The increased AFP concentration in the amniotic fluid and plasma of high-risk foals suggests upregulation.

Exfoliative cytology of human amniotic fluid (AF) has been extensively studied since 1940s, but no data exist in equine species. The AF compartment represents the environment in which the foetus grows and matures, and its composition changes, reflecting foetal well-being and development. The aim of this study was to describe for the first time the morphology of equine AF cells and amniotic membrane (AM) with light microscopy (LM) and transmission electron microscopy (TEM). AF was collected at parturition within 5 min after the appearance of the AM with a 60 mL syringe from 34 mares and samples of AM were collected from a subset of 7 mares with normal pregnancy hospitalized for attended parturition. For LM observation, a sample of cytocentrifuged fresh AF was stained with May-Grünwald Giemsa and AM sections were stained with H-E. For TEM observation, AF and AM were fixed, embedded in epoxy resins, then sectioned and stained with uranyl acetate and lead citrate solutions. Nucleated and anucleated squamous cells with basophilic cytoplasm, intensely basophilic cornified cells, polymorphonuclear cells, and clusters of eosinophilic amorphous substance were observed. Cells presumably derived from tracheal epithelium and small round nucleated cells with eosinophilic cytoplasm presumably derived from amniotic or urinary epithelium were occasionally found. Lamellar body-like structures (LBs) were present in some epithelial cells. In AM, epithelial, basal and mesenchymal layers were clearly visible with both techniques as previously described. Epithelial cells had several cytoplasmic vacuolization and microvilli were present on apical surface. The connective tissue presented fibroblasts, mesenchymal and rare polymorphonuclear cells, surrounded by abundant extracellular matrix, with distribution of collagen fibres. This is the first report about equine amniotic compartment description by LM and TEM. As recently reported in human medicine, the AM could be a second potential source of pulmonary surfactant, given the finding of LBs inside the cells which could have the same function as in humans. Further studies in samples collected at different gestational ages could increase the knowledge of AF cells and their modification during pregnancy, as well as a better comprehension of the role of AM as a secondary source of pulmonary surfactant in the horse. The diagnostic evaluation of AF cellular composition in high-risk pregnancies may also be investigated.

In the mare, foaling is a critical unpredictable event due to a wide range of gestational length and the absence of clear signs of impending parturition. To predict foaling, pH, inversion sodium potassium and increase of calcium concentration in mammary secretions are used. The aim of this study was to find how many days are left until parturition knowing mare's age (A) and parity (P) combined with ultrasonographic measurements of the fetal orbit in Standardbred mares with normal pregnancy. Eighty healthy Standardbred mares with normal pregnancy were hospitalized for attended delivery. Information about mare's age, parity and breeding date were recorded at admission. Transrectal ultrasonography were routinely performed at admission and every 10 days until parturition using a B-mode real time portable unit equipped with a 5–7.5 MHz linear transducer. The images of the fetal orbit were acquired when cornea, anterior and posterior chamber, vitreous body, lens and optic nerve were visible. Longitudinal diameter (LD) was considered as the distance between the two ocular poles, within the vitreous body; transverse diameter (TD), perpendicular to LD and bisecting the lens, was measured as the distance between cornea and retina. At delivery, length of pregnancy and gestational age at each exam were registered. For each ultrasound examination, days before parturition (DBP) were calculated. Seventy-eight Standardbred mares with normal pregnancies were included in the study. Mares' mean age was 9 ± 5 years old (range 4–20 years) and mean gestation length was 341 ± 7 days (range 327–366 days). Thirty-three mares were primiparous and 45 mares were multiparous. Data were analyzed using a regression tree: P, A, LD and TD were used as covariates. DBP was used as the variable of interest. Nine terminal nodes were identified based on the selected covariates. The first split is produced by the TD: fetuses with TD greater or equal than 2.97 cm are further split according to LD, with a threshold of 3.28 cm. The next split is dictated by A; after a further split on LD, the first terminal node is built, containing 34 fetuses with average DBP equal to 10 days. If the A is ≥ 9.5 years a further split is on P: when mares are multiparous, the TD built two different nodes. Since prediction of mare's foaling date is an important factor in stud farm management, the regression model developed may help the veterinarian to estimate the DBP in Standardbred mares with normal pregnancy.

The present study aimed to describe clinical signs of cryptosporidiosis in neonatal foals hospitalized in an Equine Perinatology Unit and to compare the clinical signs between Cryptosporidium parvum and Cryptosporidium horse genotype infection. The study was divided into two parts. In the retrospective study, nine foals infected by C. parvum were considered. In the prospective study, 70 foals, less than 15 days old, were prospectively included. Historical and clinical data were recorded, and in the prospective study, multiple fecal samples were collected. C. parvum (n = 13) and Cryptosporidium horse genotype (n = 7) were isolated. In four foals, there was a mixed infection with both the Cryptosporidium. Diarrhea, when present, showed similar duration and characteristics. Sixteen foals showed decreased abdominal sounds and colic pain before evidence of diarrhea. Nineteen foals had hyperthermia at least once. Although survival rates were similar between C. parvum (77%), C. horse genotype (100%), and cryptosporidial mixed infection (100%), foals affected by C. parvum presented anorexia (P

The matrix metalloproteinases (MMPs) are a family of enzymes involved in extracellular matrix remodeling. MMPs are secreted in a latent form and activated by local and infiltrating cells. MMP-2 and -9 are the most studied in reproduction and have been detected in bovine, ovine, equine and human placenta. There is only one study on MMPs in the equine amniotic fluid (AF) reporting a decrease in the activity of MMP-2 in case of premature delivery. The aim of this study was focused on MMP-2 and -9 activity in AF collected at parturition from mares with normal or high-risk pregnancy. High-risk pregnancy was defined as a history of premature udder development/lactation, increase of combined thickness of the uterus and placenta, vulvar discharge and/or mare's systemic illness. The diagnosis of placental insufficiency was confirmed retrospectively after macroscopic and histopatologic examination of the placenta. AF was collected by needle puncture of the amnion within 5 min after its appearance through the vulva. The activity of MMP-2 and -9 was analyzed by in-gel zymography allowing the evaluation of both latent and active forms. Twenty mares with normal pregnancy (group 1) and 8 mares with high-risk pregnancy (group 2) were included. All mares in group 2 had a high-risk pregnancy with a diagnosis of placental insufficiency associated with placental villous hypoplasia, placentitis or placental edema. The bands relative to latent and active forms of MMP-2 were clearly visible in both groups and the activity of latent (P = 0.010) and active (P = 0.004) forms was lower in the AF samples of group 2. The band of the latent form of MMP-9 was visible in 17/20 samples of group 1, while it was completely absent in all samples of group 2. In contrast, the band of the active form was clearly visible and with a greater activity in AF samples of group 2 (P = 0.002). Placental dysfunction seems to induce a lower MMP-2 activity and a higher MMP-9 activity through the release of pro-inflammatory cytokines. Because fetal pulmonary secretions are a likely source of gelatinases in AF during late gestation, the increased MMP-9 activity could be related to fetal distress. These data provide a starting point to better understand the role of MMPs in equine pregnancy, although it should be confirmed in a larger and more homogeneous population of mares with high-risk pregnancy.

Simple Summary Post-antibiotic intestinal dysbiosis leads to an overall reduction in bacterial and functional diversity, along with a minor resistance against pathogens. The study aimed to determine the changes on the fecal microbiota in hospitalized neonatal foals administered with broad-spectrum antimicrobials and supplemented probiotics. Fecal samples were collected at hospital admission, at the end of the antimicrobial treatment and at discharge. Seven foals treated with intravenous ampicillin and aminoglycosides for a mean of seven days were included. The results suggest that the fecal microbiota of neonatal foals rapidly returns to a high diversity after treatment. While the findings need to be confirmed in a larger population, the study suggests that in foals, the effect of antimicrobials may be strongly influenced by the changes that occur over time in the developing gut microbiota. Of note, the findings are influenced by the use of probiotics, and whether the changes would be consistent in antimicrobial-administered but not supplemented foals remains to be elucidated. Abstract There is a wide array of evidence across species that exposure to antibiotics is associated with dysbiosis, and due to their widespread use, this also raises concerns also in medicine. The study aimed to determine the changes on the fecal microbiota in hospitalized neonatal foals administered with broad-spectrum antimicrobials and supplemented probiotics. Fecal samples were collected at hospital admission (Ta), at the end of the antimicrobial treatment (Te) and at discharge (Td). Feces were analysed by next-generation sequencing of the 16S rRNA gene on Illumina MiSeq. Seven foals treated with IV ampicillin and amikacin/gentamicin were included. The mean age at Ta was 19 h, the mean treatment length was 7 days and the mean time between Te and Td was 4.3 days. Seven phyla were identified: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, TM7 and Verrucomicrobia. At Ta, Firmicutes (48.19%) and Proteobacteria (31.56%) were dominant. The alpha diversity decreased from Ta to Te, but it was the highest at Td. The beta diversity was higher at Ta than at Te and higher at Td than at Te. An increase in Akkermansia over time was detected. The results suggest that the intestinal microbiota of neonatal foals rapidly returns to a high diversity after treatment. It is possible that in foals, the effect of antimicrobials is strongly influenced or overshadowed by the time-dependent changes in the developing gut microbiota.

Hydramnios is an excessive accumulation of fluid within the amniotic compartment. It is a rare condition in mares, often associated with fetal anomalies. Hydrops of fetal membranes predisposes to the rupture of the prepubic tendon, and many authors suggest the induction of parturition to preserve mare’s reproductive career. This report presents the case of a 15-year-old multiparous Quarter Horse mare, referred at 268 days of gestation for suspected hydrops. Repeated ultrasonographic exams confirmed an increase in the depth of the amniotic fluid and reduced fetal viability. During the hospitalization, the mare developed a partial rupture of the prepubic tendon. In this case, a conservative approach was elected, and the mare was treated with nonsteroidal anti-inflammatory drugs (NSAIDs) and an abdominal support bandage. At 327 days of gestation, the mare gave birth to a foal with APGAR score 1. The resuscitation attempt was unsuccessful, and the foal died immediately. A post-mortem examination diagnosed a congenital diaphragmatic hernia (CDH) with pleuroperitoneal diaphragmatic eventration.

Background Ultrasonographic measurement of the vena cava and aorta (Ao) diameters and their ratios have been suggested to be a reliable way of quantifying hypovolemia. Objective To evaluate the feasibility and reliability of an ultrasonographic technique for measurement of Ao and caudal vena cava (CVC) and derived ratios using three different acoustic windows in a population of healthy neonatal foals. Correlation between Ao and CVC measurements and ratios and foals' age or bodyweight were also investigated. Methods In 14 healthy foals aged less than 7 days, the diameters of the Ao and of the CVC in long and short axis were measured by two observers from images obtained using three different ultrasonographic imaging planes (left dorsal, left ventral and right views). The Ao and CVC cross‐sectional area and the CVC/Ao diameter and area ratios were calculated. Image quality was subjectively assessed. Intraobserver and interobserver reliabilities for image quality scores and measurements were evaluated between the two observers. Simple linear regression models were used to identify correlations between the CVC/Ao measurements and ratios and the age and bodyweight of the foals. Results The left ventral view showed the highest reliability. A correlation between bodyweight and the short axis measurement of the CVC was found (R 2 = 0.385; p = 0.018). Age was positively correlated with the long axis of measurement of the CVC (R 2 = 0.426; p = 0.011) and CVC/Ao diameter ratio (R 2 = 0.625; p = 0.001). Conclusions The left ventral view allows the Ao and CVC cross sections to be easily visualized and measured in neonatal foals in right lateral recumbency., The present study evaluated the feasibility and reliability of an ultrasonographic technique for measurement of Ao and CVC and derived ratios using three different acoustic windows in a population of healthy neonatal foals. A left ventral view of the abdomen allowed for rapid imaging and measurement of the cross‐sectional diameter of both the aorta and the CVC in neonate foals in lateral recumbency, and might be a promising method to quantitatively assess the degree of hypovolemia.

A complex feedback of growth factors, secreted by a variety of cell types, is responsible for the mediation of skin healing. Despite the recent advances in wound healing management, this fails up to 50% and skin wounds can still be considered one of the main causes of morbidity, both in human and veterinary medicine. Regenerative medicine, involving mesenchymal stromal cells (MSCs), is nowadays a promising solution for skin wound healing. Indeed, MSCs are involved in the modulation of the inflammatory local response and cell replacing, by a paracrine mode of action. Local application of equine umbilical cord Wharton's jelly MSCs (WJMSCS) was carried out in a 6-months-old filly with a non-healing skin wound. Heterologous WJMSCs were applied four times using a carboxymethylcellulose (CMC) gel, produced dissolving CMC in autologous plasma. At first application the mean wound area was 7.28 ± 0.2 cm2. Four days after the last application of WJMSCs, the mean wound area was 1.90 ± 0.03 cm2, and the wound regression rate was +74%. No local or systemic side effects were registered after WJMSCs application and no evident exuberant scar was observed after wound healing. At discharge, the mean wound area was 0.38 ± 0.01 cm2 and the total regression rate was +80%. Five days later, the wound was completely healed. In the present clinical case report, the use of WJMSCs led to promising clinical results, paving the way for possible future applications in the treatment of chronic wounds in horses.

Background: Blood collection by indwelling intravenous catheter (IVC) avoids repeated venipuncture, which could cause thrombophlebitis risk, anxiety and pain in patients. Objectives: To compare blood gas parameters, electrolytes, glucose, lactate and haematocrit concentration obtained from venous blood samples collected via a jugular IVC by push-pull (PP) technique to those obtained by venipuncture in hospitalised foals, at the time of catheter placement (T0) and 24hours after the beginning of intravenous therapy (T24). Study design: Prospective observational study. Methods: Paired blood samples were drawn from hospitalised foals at T0 and T24. In each foal, one venous blood sample was collected via IVC by the following PP technique: 2.4mL of blood was aspirated and immediately reinfused through the catheter three times consecutively, then 1mL of blood was collected using a 1mL heparinised syringe. Thereafter, another sample was collected by direct venipuncture of the contralateral jugular vein, with an identical 1mL heparinised syringe, with a 1-inch, 20-G needle. All samples were analysed with an automated blood gas analyser within 10minutes of collection. The agreement between the two techniques was assessed by Bland-Altman analysis and intraclass correlation coefficient (ICC). Results: The level of agreement of blood gas values obtained by the two different techniques was high with very small bias and clinically acceptable ICC (>0.907 at T0; >0.794 at T24) for all variables, except for haematocrit (bias −3.52 at T0; −2.44 at T24) and PvO2 at T0 and T24 (ICC 0.669 and 0.733, respectively). Main limitations: Potential sub-clinical catheter-related complications were not investigated by ultrasound or bacterial culture of the catheter; short duration of the study. Conclusions: PP technique appears to be acceptable for collection of blood samples for venous blood gas parameters, as well as electrolytes, glucose and lactate in sick neonatal foals.

Induction of parturition in horses is still not well accepted due to the potential peripartum complications for mares and newborn foals. We assessed differences after spontaneous and induced parturition with low doses of oxytocin (OX) in 1) incidence of peripartum complications in mares; 2) viability, behavioral, physical, and venous blood gas analyses in foals. In this study 61 mares were included; 45/61 were enrolled in the spontaneous foaling group (SF) and 16/61 in the induced foaling group (IF). In the IF group, when the calcium in mammary secretion reached concentrations of ≥250 ppm, mares received a single injection of 2.5 IU of oxytocin IV once a day until foaling. Mares’ breed, age, parity, gestational and stage II length, and peripartum complications were recorded. Foal maturity, vital (Apgar score), behavioral and physical parameters were assessed at birth, and the foal clinical condition was monitored for one week. A jugular venous blood sample was collected at birth for blood gas analysis, acid-base status, and lactate assessment. The median gestational length was within the reference interval in all the mares included and did not differ between the two groups. No statistical differences in the II stage length nor in incidence of peripartum complications were observed between the two groups. All the foals were born alive and showed no signs of prematurity/dysmaturity. No statistical differences were found in foal viability between the two groups. Time to stand and nurse from the mare, and body temperature were significantly higher in the IF compared to the SF group. Venous blood pH, SO2% and BE were lower, while pCO2 and lactate were higher in the IF than in the SF group. All the foals in both groups remained clinically healthy during the observation period. In conclusion, at term induction of parturition with a low dose of oxytocin does not have adverse effects on peripartum in mares. Our findings suggest that at term induced foals suffer slightly greater, but not clinically significant, hypoxia, hypercapnia and acidosis than spontaneously delivered foals.

Wharton's jelly (WJ) is an important source of mesenchymal stem cells (MSCs) both in human and other animals. The aim of this study was to compare human and equine WJMSCs. Human and equine WJMSCs were isolated and cultured using the same protocols and culture media. Cells were characterized by analysing morphology, growth rate, migration and adhesion capability, immunophenotype, differentiation potential and ultrastructure. Results showed that human and equine WJMSCs have similar ultrastructural details connected with intense synthetic and metabolic activity, but differ in growth, migration, adhesion capability and differentiation potential. In fact, at the scratch assay and transwell migration assay, the migration ability of human WJMSCs was higher (P

Botulism, a severe neuroparalytic disease that can affect humans, all warm-blooded animals, and some fishes, is caused by exotoxins produced by ubiquitous, obligate anaerobic, spore-forming bacteria belonging to the genus Clostridium and named botulinum neurotoxin (BoNT)-producing clostridia. This report presents the case of a 3-year-old donkey mare referred for progressive and worsening dysphagia of four days’ duration. Her voluntary effort in eating and drinking was conserved, and she was able to slow chew without swallowing. A complete neurological examination was performed, and botulism was strongly suspected. The ability to swallow feed and water returned on the tenth day of hospitalization and improved progressively. The jenny was discharged from the hospital after fifteen days. During the hospitalization, the Italian National Reference Centre for Botulism confirmed the diagnosis: mare’s feces were positive for BoNT/B and Clostridium botulinum type B.

A complex feedback of growth factors, secreted by a variety of cell types, is responsible for the mediation of skin healing. Despite the recent advances in wound healing management, this fails up to 50% and skin wounds can still be considered one of the main causes of morbidity, both in human and veterinary medicine. Regenerative medicine, involving mesenchymal stromal cells (MSCs), is nowadays a promising solution for skin wound healing. Indeed, MSCs are involved in the modulation of the inflammatory local response and cell replacing, by a paracrine mode of action. Local application of equine umbilical cord Wharton's jelly MSCs (WJMSCS) was carried out in a 6-months-old filly with a non-healing skin wound. Heterologous WJMSCs were applied four times using a carboxymethylcellulose (CMC) gel, produced dissolving CMC in autologous plasma. At first application the mean wound area was 7.28 ± 0.2 cm2. Four days after the last application of WJMSCs, the mean wound area was 1.90 ± 0.03 cm2, and the wound regression rate was +74%. No local or systemic side effects were registered after WJMSCs application and no evident exuberant scar was observed after wound healing. At discharge, the mean wound area was 0.38 ± 0.01 cm2 and the total regression rate was +80%. Five days later, the wound was completely healed. In the present clinical case report, the use of WJMSCs led to promising clinical results, paving the way for possible future applications in the treatment of chronic wounds in horses.

Background Little is known about the differences among adult and foetal equine mesenchymal stem cells (MSCs), and no data exist about their comparative ultrastructural morphology. The aim of this study was to describe and compare characteristics, immune properties, and ultrastructural morphology of equine adult (bone marrow: BM, and adipose tissue: AT) and foetal adnexa derived (umbilical cord blood: UCB, and Wharton's jelly: WJ) MSCs. Results No differences were observed in proliferation during the first 3 passages. While migration ability was similar among cells, foetal MSCs showed a higher adhesion ability, forming smaller spheroids after hanging drop culture (P < 0.05). All MSCs differentiated toward adipogenic, chondrogenic and osteogenic lineages, only tenogenic differentiation was less evident for WJ-MSCs. Data obtained by PCR confirmed MHC1 expression and lack of MHC2 expression in all four cell types. Foetal adnexa MSCs were positive for genes specific for anti-inflammatory and angiogenic factors (IL6, IL8, IL beta 1) and WJ-MSCs were the only positive for OCT4 pluripotency gene. At immunofluorescence all cells expressed typical mesenchymal markers (alpha-SMA, N-cadherin), except for BM-MSCs, which did not express N-cadherin. By transmission electron microscopy, it was observed that WJ-MSCs had a higher (P < 0.05) number of microvesicles compared to adult MSCs, and UCB-MSCs showed more microvesicles than BM-MSCs (P < 0.05). AT-MSCs had a lower number of mitochondria than WJ-MSCs (P < 0.05), and mitochondrial area was higher for WJ-MSCs compared to UCB and AT-MSCs (P < 0.05). Conclusions Results demonstrate that MSCs from adult and foetal tissues have different characteristics, and foetal MSCs, particularly WJ derived ones, seem to have some charactestics that warrant further investigation into potential advantages for clinical application.

Background Even if horses strictly depend on the gut microbiota for energy homeostasis, only a few molecular studies have focused on its characterisation and none on the perinatal gut microbial colonisation process. Objectives To explore the perinatal colonisation process of the foal gut microbial ecosystem and the temporal dynamics of the ecosystem assembly during the first days of life. Study design Longitudinal study. Methods Thirteen Standardbred mare-foal pairs were included in the study. For each pair, at delivery we collected the mare amniotic fluid, faeces and colostrum, and the foal meconium. Milk samples and faeces of both mare and foal were also taken longitudinally, until day 10 post-partum. Samples were analysed by means of next-generation sequencing of the 16S rRNA gene on Illumina MiSeq. Results Our findings suggest that microbial components derived from the mare symbiont communities establishes in the foal gut since fetal life. After birth, an external transmission route of mare microorganisms takes place. This involves a rapid and dynamic process of assembling the mature foal gut microbiome, in which the founder microbial species are derived from both the milk and the gut microbial ecosystems of the mare. Main limitations The inability to discriminate between live and dead cells, the possible presence of contaminating bacteria in low biomass samples (e.g. meconium and amniotic fluid), the limits of the phylogenetic assignment down to species level, and the presence of unassigned operational taxonomic units. Conclusions Our data highlight the importance of mare microbiomes as a key factor for the establishment of the gut microbial ecosystem of the foal.

none 4 no This article describes diagnosis, treatment, surgical management, and outcome in 16 foals (mean age 7 days of life) affected by confirmed tarsocrural septic arthritis. The chosen therapy was lavage of the synovial cavity performed arthroscopically, once or twice due to recurrence of clinical signs. Information recorded included signalment, history, a complete clinical evaluation, grade of lameness, hematology, biochemistry, serum immunoglobulin G concentration, bone infection radiological findings, cytology and culture of the synovial fluid, treatment, and outcome. Eleven of the foals were males (69%), and five were females (31%). Sepsis affected the right tarsocrural joint in three foals (19%) and the left one in seven foals (44%). Six foals had both hocks affected (37%). In 12 foals (75%), the infection was successfully resolved, and a single intervention was sufficient to have remission of symptoms and resolution of infection. Arthroscopic lavage was repeated in four of 16 foals (25%); five /16 foals (31%) had two joints involved, and the arthroscopic lavage was repeated for all joints. Although the actual literature describes poor possibilities to explore joints in the foal, in this case, it was possible to visualize the inner structures, correctly ascertain and grade the intra-articular damage resulting from infection, and the presence of organic materials inside the joint space. Moreover, what is worth noting is that it was possible to detect and remove fibrin clots thought to be responsible for hiding bacteria and carrying on the infective and inflammatory status. none Rinnovati, Riccardo*; Butina, Barbara Bianchin; Lanci, Aliai; Mariella, Jole Rinnovati, Riccardo*; Butina, Barbara Bianchin; Lanci, Aliai; Mariella, Jole

The umbilical cord (UC), the connection between mother and fetus via the umbilical vessels, carries nutrients and oxygenated blood to the fetus through the umbilical vein and removes deoxygenated blood and waste products via the umbilical arteries. It is designed to protect blood flow to the fetus during pregnancy. In equine medicine, only a few studies have described the UC, and most of these involved Thoroughbreds. The present study describes and compares the macroscopic features of the equine umbilical cord in three different breeds and in relation to the foal's gender. In addition, a possible correlation between UC features and maternal and perinatal factors is investigated. One hundred and twenty four healthy mares with normal pregnancies were enrolled in the study and were divided into three groups according to their breed: 70 Standardbreds (STB), 38 Thoroughbreds (THB) and 16 Warmbloods (WAB). The following data were recorded: mare's age and parity, gestation length, placental weight, presence of fetal membrane alterations, UC length and number of coils in the amniotic and allantoic portions, and the Umbilical Coiling Index (UCI), which is the ratio between total coils and total UC length. The UCI has not been investigated previously in veterinary medicine. Furthermore, immediately after foaling, APGAR score, foal's weight and sex were recorded. All the STB and WAB were housed in Italy and the THB were housed in New Zealand. Mares' mean age was higher in WAB than in THB and STB; the latter had a significantly shorter gestation length. The foal's weight was positively correlated with placental weight in all breeds; and in STB, foal weight was positively related to parity and gestation length. Mean total UC length was comparable to previous reports in THB, STB and WAB. The lengths of the two UC portions were statistically different between STB and THB, where the amniotic portion was longer than the allantoic one. In each breed, total UC length was correlated with total number of coils (THB and STB = 5 ± 1; WAB = 6 ± 1), the UC amniotic length was positively correlated with the number of amniotic coils and the allantoic length was positively correlated with the number of allantoic coils. The UCI values were 0.09 in STB and THB and 0.1 in WAB. This study provides reference values for UCI that could be included in the gross placental evaluation if its clinical importance were demonstrated.

Both in human and equine species, mesenchymal stem cells (MSCs) from amniotic membrane (AM) and Wharton’s jelly (WJ), may be particularly useful for immediate use or in later stages of life, after cryopreservation in cell bank. The aim of this study was to compare equine AM- and WJ-MSCs in vitro features that may be relevant for their clinical employment. MSCs were more easily isolated from WJ, even if MSCs derived from AM exhibited more rapid proliferation (P P

Placental changes associated with artificial reproductive technologies have been described in several species, but little information is available in horses. Joy et al. (2012) reported that human placentas from intracytoplasmic sperm injection derived embryos were heavier and thicker than those produced after natural conception. Despite the most growing interest and efficiency of artificial reproductive technologies in equine species, only recently, Pozor et al. (2016) described placental abnormalities in pregnancies generated by somatic cell NT, but there are no studies on equine placenta generated by intracytoplasmic sperm injection and traditional embryo transfer. In the present preliminary study, macroscopic differences of placentas generated after transfer of in vitro- or in vivo-produced embryos were registered. Twelve Standardbred recipient mares with pregnancy generated after transfer of in vivo-derived (Group 1) and in vitro-derived (Group 2) embryos were enrolled; 10 Standardbred mares with pregnancy derived by traditional AI were included as control (Group 3). All pregnancies were physiological, and newborn foals were healthy. Mare age, parity, length of pregnancy, gross evaluation and weight of placenta, total length of umbilical cord (UC), length of UC, number of UC coils, foal sex, and weight at birth were registered. Collected data are listed in Table 1 and are expressed as mean ± standard deviation. Differences between groups were evaluated by 1-way ANOVA, and the difference in proportion of overweight placentas was evaluated with the Fisher test. The gross evaluation of placenta revealed 8/12 placentas (2/4 Group 1; 6/8 Group 2) were heavier than 11% (Madigan, 1997) due to oedema of the chorioallantois. No overweight placentas were registered in Group 3. In Group 1, 1/4 placentas had villous hypoplasia, and in Group 2, 1/8 placentas had cystic pouches on the UC. There were no significant differences among groups. However, the proportion of overweight placentas between Group 2 (6/8) and Group 3 (0/10) approached significance (P = 0.06). Although preliminary, the results of the present study suggest that production of equine embryos in vitro may lead to alterations in placental development. Several studies in cattle and sheep have suggested that alterations in the placentas of pregnancies derived from in vitro-produced embryos are related to effects of culture on epigenetic regulation. Less is known in the horse about the effects of in vitro embryo production on placental development; thus, further research in this area is necessary. Table 1. Characteristics of full-term placentas derived from AI or embryo transfer with in vivo- and in vitro-produced embryos