Healthy Mares Research Articles

Molecular investigation of the prevalence of Equine alphaherpesvirus‐1 infection in healthy postpartum mares

Molecular investigation of the prevalence of Equine alphaherpesvirus‐1 infection in healthy postpartum mares

Clinical Outcome of Transcervical Infusion of a Combination of Procaine Penicillin and Gentamicin in Late-term Pregnant Mares

Transcervical intrauterine infusion of antibiotics may more effectively treat pathogens associated with fetal and neonatal disease in pregnant mares than standard systemic routes. The objective of this study was to assess the safety of transcervical antibiotic infusion by characterizing the gestational outcome in nine healthy pregnant pony mares following a single transcervical infusion of 2.4 million IU of procaine penicillin and 200 mg of gentamicin in a 10 mL volume during late gestation. Assessment of fetal-placental health was performed through serial measurement of the combined thickness of the uterus and placenta (CTUP) and fetal heart rate and mares and foals were closely monitored in the periparturient period. Fetal heart rate and CTUP remained unchanged after infusion, with no evidence of fluid accumulation or significant increase at the time-points 24, 48, and 72 hours. All mares foaled without complication 12–58 days after antibiotic infusion at a mean gestational age of 322.7 ± 12.7 days. Two out of nine foals displayed signs of mild neonatal maladjustment syndrome that responded to minimal supportive care and all foals survived to weaning without further complications.

Three Manual Noncommercial Methods to Prepare Equine Platelet-Rich Plasma.

Simple SummaryPlatelet-rich plasma (PRP) is a popular therapy in human and veterinary medicine. This study compared three manual noncommercial methods to prepare PRP and its cooling. The three methods concentrated platelets and did not alter viability. Method 1 (i.e., involving double centrifugation) resulted in the greatest platelet concentrations, while method 3 (sedimentation) resulted in the lowest platelet concentration and greater contamination with white blood cells (WBC). Cooling increased platelet agglutination over time across methods and affected platelet viability in PRP obtained with method 3. It remains unknown whether the different methods and cooling would affect PRP’s clinical efficacy.In light of PRP’s increasing popularity in veterinary practice, this study aimed to compare three manual methods to prepare and cool equine PRP. The blood of 18 clinically healthy mares was collected via venipuncture in a blood transfusion bag (method 1), blood tubes (method 2), and a syringe (method 3). In method 1, samples were double centrifuged; method 2 involved one centrifugation, and in method 3 the syringe was kept in an upright position to sediment for 4 h. After processing with three methods, PRP and platelet-poor plasma (PPP) were extracted and assessed for red (RBC) and white blood cell counts (WBC), platelet counts, and viability. In a subset of mares (n = 6), samples were processed with the three methods, and PRP was evaluated at 6 and 24 h postcooling at 5 °C. Method 1 resulted in the highest and method 3 in the lowest platelet concentration (p < 0.05), and the latter also had greater contamination with WBC than the others (p < 0.001). Platelet viability was similar across treatments (p > 0.05). Cooling for 24 h did not affect platelet counts in all methods (p > 0.05); however, platelet viability was reduced after cooling PRP produced by method 3 (p = 0.04), and agglutination increased over time in all methods (p < 0.001). The three methods increased (1.8–5.6-fold) platelet concentration in PRP compared to whole blood without compromising platelet viability. In conclusion, all three methods concentrated platelets and while cooling affected their viability. It remains unknown whether the different methods and cooling would affect PRP’s clinical efficacy.

Hemodynamic changes in cytokines, chemokines, acute phase proteins and prostaglandins in mares with subclinical endometritis

Apparently healthy mares with conception failure or embryonic loss may have subclinical endometritis (SE). Our objective was to document evidence of systemic immune responses against SE in mares. In apparently healthy (control) mares as well as those with chronic endometritis (ChE) or subacute suppurative endometritis (SSE), both considered classes of SE, serum concentrations of cytokines (IL-1β, IL-6, IL-10 and TNF-α), chemokines (IL-8), acute phase protein (SAA), and plasma concentrations of prostaglandins (PGF2α and PGE2) were measured using validated enzyme linked immunoassays (EIA). Mixed-breed mares of known reproductive history, were used. Based on an endometrial cytological examination, mares were allocated into the following groups: healthy (control), ChE, and SSE (12, 26 and 11 mares, respectively). Serum concentrations of IL-6, IL-8 and IL-10 and plasma PGF2α concentrations were increased (P < 0.01) in mares with SSE compared to healthy mares. Furthermore, serum concentrations of IL-6 (P < 0.05) and IL-10 (P < 0.05) were elevated in the ChE group compared to the healthy group whereas serum SAA (P < 0.01) and plasma PGE2 (P < 0.05) were higher in healthy mares compared to ChE and SSE mares. However, serum concentrations of IL-1β and TNF-α were not significantly different among groups. In conclusion, there were two novel findings: (1) development of serum testing for a set of biochemical markers has promise to explore the pathogenesis of inflammation in mares with SE; and (2) none of the individual biomarkers studied, excluding IL-8, was a significant predictor of SE. However, serum IL-6:IL-10 and IL-6:TNF-α concentrations, as well as plasma PGE2:PGF2α ratios, may yield a novel diagnostic marker for chronic subclinical endometritis.

Effect of topical cyclopentolate alone or combined with phenylephrine in healthy horses.

Evaluate the effect of repeated doses of topical 1% cyclopentolate hydrochloride alone and in combination with topical 2.5% phenylephrine on pupil diameter (PD), tear production (STT-1), intraocular pressure (IOP), digestive function (gut motility and feces production), and heart rate (HR). Six healthy mares. In a prospective, randomized, controlled, and crossover design study, the left eye of six healthy mares was administered 0.2mL of cyclopentolate alone and in combination with 0.2mL of phenylephrine. The drugs were administered 3 times a day for 1day, twice a day for 1day, and then once a day for 2days, as commonly used in practice. Daily and two days after the last topical drug administration, HR, digestive auscultation, feces production, STT-1, IOP, and PD were recorded. The cyclopentolate alone significantly increased the horizontal and vertical PD of the treated eye from day 2 to day 6 (p<.0001) compared with the baseline value. The combination with topical phenylephrine did not have any additional effect on mydriasis compare with the cyclopentolate alone. The other ocular and digestive parameters were not affected by repeated doses of cyclopentolate alone or combined. Repeated administration of cyclopentolate alone or combined with phenylephrine induce a significant mydriasis for at least 48h after the last administration in normal horses' eyes, and do not affect STT-1, IOP, digestive function, and HR. The phenylephrine combined with the cyclopentolate did not potentiate the pupil dilation when compared with cyclopentolate alone in healthy horses.

109 Is the equine gut colonised in utero by amniotic bacteria?

Until very recently it was believed that the uterus was sterile, with microbial colonisation of the foal's gastrointestinal tract (GIT) commencing at birth. GIT colonisation is an integral developmental process, critical for an optimal functioning immune system and ongoing health of the foal. The in utero colonisation paradigm has now challenged this concept, indicating that the amniotic fluid (AF) composition contains non-pathogenic microbiota that potentially commence colonisation of the GIT during the process of fetal swallowing. In turn, preparing the foal for life outside the womb. This study examined the presence of microbiota in the AF of healthy pregnant mares, identifying microorganisms that have the potential to colonise the fetal GIT in utero. AF samples were obtained from 14 healthy mares in the second stage of parturition, before the rupture of the amnion. When the amnion was clearly visible, the surface was seared with a sterile spatula and AF extracted with a sterile needle and syringe. The sample had no contact with any other surface of the mare and was immediately snap-frozen and stored at −80°C for microbiota analysis. Genomic DNA was extracted and processed for 16S rRNA gene amplicon sequencing, followed by bioinformatic analysis of the results using QIIME2 and CalypsoV8.84 software packages. Microbiota was detected in all AF samples. Communities were of low diversity, as indicated by the α diversity parameters Chao1, Evenness, Richness and Simpson's Index. Operational Taxonomic Units (OTUs) of the top 20 taxa showed some host variation. The most abundant OTUs across the majority of samples were classified to the phyla Opisthokonta (100% of samples), Bacteroidetes (79%), Actinobacteria (57%) and Firmicutes (57%). At the family level, Staphylococcacea (79%) and Microbacteriaceae (71%) were the most abundant. These microbes have been previously detected across other locations of the equine microbiome, including the oral, esophageal, distal GIT and vaginal ecosystems. Our results have demonstrated the presence of a complex microbial ecosystem of low diversity in the AF of healthy equine pregnancies, likely of maternal origin. The environmental and gastrointestinal microbes detected in the AF displayed similarities to other equine ecosystems, suggesting microbial translocation from the mare to the AF. Further research would be required to determine if these AF microbes are being ingested by the fetus, thus commencing GIT colonisation in utero . Deciphering microbial colonisation of the foal's GIT has important implications for maternal management before parturition and could lead to the development of strategies to enhance foal health outcomes.

Epidemiology of Chlamydia psittaci infections in pregnant Thoroughbred mares and foals

Late-term foal loss due to the traditional avian pathogen Chlamydia psittaci recently emerged as a threat to the Australian Thoroughbred industry. A longitudinal study of 14 stud farms was undertaken to better understand C. psittaci infection in pregnant mares and their foals by evaluating C. psittaci prevalence, equine herpesvirus-1 (EHV-1) co-infection, avian reservoirs, and potential risk factors. Mucosal swabs taken from 228 healthy pregnant mares and their foals were tested for C. psittaci and EHV-1 using species-specific qPCR assays.No foal loss was recorded due to either pathogen, and no mare tested positive to either C. psittaci or EHV-1. However, healthy newborn foals tested positive to both pathogens, at low levels, with 13.2% (n = 30/228) and 14.5% (n = 33/228) prevalence for C. psittaci and EHV-1, respectively. Co-infection occurred in 1.3% (n = 3/228) of foals. In avian environmental faecal samples collected from the same studs, C. psittaci was detected at 5.3% (n = 5/94). Multiple logistic regression modelling found that foals born in winter were more likely to be infected with C. psittaci (adjusted odds ratio = 15.83; P < 0.001; Confidence Interval 5.12–48.49). Being a maiden mare, absence of prophylactic vaginal suture, interventions in the last trimester and residing on a farm with prior history of C. psittaci abortion posed no higher risk to infection in the newborn. Analysis of all reported C. psittaci abortion cases (Hunter Valley, 2016–2019) revealed a dominant C. psittaci sequence type (denoted ST24) and a significant correlation with frost events (Spearmans’ rho = 0.44; P = 0.002).

Expression of Enzymes Associated with Prostaglandin Synthesis in Equine Conceptuses.

Simple SummaryThe mobile preimplantative phase of equine gestation, taking place between day 9 and 16 after ovulation, is characterized by peristaltic contractions of the uterus caused by secretion of prostaglandins by the spheric equine conceptus. This mobility is necessary for maternal recognition of pregnancy in equids, taking place around day 14 after ovulation. The presented study investigated the spatial and temporal abundance of prostaglandin synthesis enzymes of the equine conceptus, elucidating a basal and an inducible system for prostaglandin E2. Prostaglandin F2α synthesis is restricted to the “periembryonic”pole area and relies on enzymatic conversion of prostaglandin E2. This scenario led to a model able to explain the embryonic forward motion driven by the peristaltic contractions of the uterus. In vitro incubation of primary trophoblast cell cultures with oxytocin showed no influence of this hormone on prostaglandin synthesis.In the horse, mobility of the conceptus is required for maternal recognition of pregnancy depending on secretion of prostaglandins by the conceptus. The aim of this study was to determine the expression and localization of key enzymes of the different pathways leading to synthesis of prostaglandin E2 and F2α in the equine conceptus during the mobility phase. Enzyme expression was analyzed via quantitative RT-PCR in total RNA samples of equine conceptuses collected on days 10 (n = 5), 12 (n = 12), 14 (n = 5) and 16 (n = 7) from healthy mares. Relative abundance of cyclooxygenase (COX)-2 mRNA was higher (p < 0.05) than of COX-1 irrespective of conceptus age and for phospholipase A2 on day 16 in comparison to all other days (p < 0.01). Abundance of mRNA of cytosolic and microsomal prostaglandin E synthase (PGES) and of carbonyl reductase (CBR) 1 was not influenced by conceptus age. Immunohistochemically, COX-1, COX-2, as well as cytosolic and microsomal PGES were present in both the ectodermal and endodermal layer of the yolk sac wall. CBR-1 was restricted to periembryonic disc area. The localisation of the key enzymes explains the mechanism of embryo mobility. In vitro incubation of primary trophoblast cell cultures with oxytocin had no effect on key enzyme synthesis.

Comparison of sedation quality and safety of detomidine and romifidine as a continuous rate infusion for standing elective laparoscopic ovariectomy in mares

To compare efficacy and safety of a continuous rate infusion of detomidine hydrochloride and romifidine hydrochloride for standing elective bilateral laparoscopic ovariectomy in mares. Blinded, randomized prospective clinical study. Eighteen healthy mares presenting for elective bilateral ovariectomy METHODS: Mares were randomly assigned to one of two sedation protocols. Prior to surgery, baseline head height, heart rate, respiratory rate, and postural sway were recorded. An IV loading dose of α2-agonist (46 μg/kg romifidine or 13.9μg/kg detomidine) was administered. Standing sedation was maintained with a continuous rate infusion of the respective α2-agonist (126 μg/kg/h romifidine or 37.8μg/kg/h detomidine). Intraoperative measurements included respiratory rate, heart rate, head height, postural sway, and response to surgical stimulus. Postoperatively, fecal output was recorded, and pain scoring was performed using composite pain score and visual analog scales. Three of 18 horses required additional α-2 agonists: one detomidine and two romifidine and butorphanol. Head height during surgery was lower (p < .001) in mares receiving detomidine. Postural sway around the vertical axis was greater in mares sedated with detomidine rather than romifidine (p=.013). No differences were detected in intraoperative heart rate, postoperative pain scores or postoperative fecal output between sedation techniques. Comparable scores for surgical stimulation and sedation were measured in both sedation groups. No differences in postoperative analgesia or manure production were identified. Romifidine appears suitable as an alternative to detomidine and may limit ataxia and head drop in sedated horses.

Successful cardiopulmonary resuscitation in an adult horse following cardiovascular collapse on recovery from general anaesthesia in the Trendelenburg position

SummaryEquine anaesthesia is associated with a high risk of perioperative morbidity and mortality, particularly in the recovery period. Cardiovascular collapse and cardiopulmonary arrest (CPA) have been reported to account for one‐third of anaesthesia‐related perioperative deaths in the horse. This case report describes the successful cardiopulmonary resuscitation of a healthy adult Thoroughbred mare that developed cardiovascular collapse and subsequent CPA following positioning in the recovery box after general anaesthesia in the Trendelenburg position. Cardiopulmonary resuscitation (CPR) was instigated and included thoracic compressions, intermittent positive pressure ventilation and adrenaline administered via the intravenous and intra‐tracheal routes. Return of spontaneous circulation (ROSC) was detected 5 min and 50 s after the start of CPR. A stable stance was achieved approximately 100 min after ROSC. The mare was discharged from the hospital 3 days later with no known complications. To the authors’ knowledge, this is the first case report documenting CPA following general anaesthesia in the Trendelenburg position in a horse, with subsequent successful CPR. The precise cause of the CPA is unknown but a number of plausible hypothesis are discussed including hypoventilation and a Bezold‐Jarisch reflex.

Changes Associated with the Peri-Ovulatory Period, Age and Pregnancy in ACTH, Cortisol, Glucose and Insulin Concentrations in Mares.

Simple SummaryThe reproductive cycle of mares is associated with many hormonal changes, but the effects of this cycle and pregnancy on adrenocorticotropic hormone and insulin concentrations are poorly described, which could limit our ability to diagnose pituitary pars intermedia dysfunction and insulin dysregulation. In this study, healthy mares are followed through their reproductive cycle during the physiologic breeding season, and adrenocorticotropic hormone, cortisol, glucose and insulin concentrations are measured to determine the effects of age, pregnancy and the reproductive cycle on analyte concentrations. No significant effect of age, the reproductive cycle or pregnancy is detected on the cortisol, glucose or insulin concentrations; however, adrenocorticotropic hormone concentrations are significantly increased throughout the peri-ovulatory period and during pregnancy. Therefore, knowledge of a mare’s reproductive cycle might be beneficial when interpreting adrenocorticotropic hormone concentrations.Although there are many hormonal changes associated with reproduction, the effects of ovulation and early pregnancy on adrenocorticotropic hormone (ACTH) and insulin concentrations are poorly described. We hypothesise that both ovulation and early pregnancy will alter ACTH and insulin concentrations in healthy mares. Eighteen mares showing no clinical signs suggestive of, or laboratory findings consistent with, pituitary pars intermedia dysfunction PPID and insulin dysregulation (ID) are enrolled. ACTH, cortisol, insulin and glucose concentrations are measured over their peri-ovulatory period, as determined via ultrasounds and progesterone concentrations. The mares are grouped by age and gestation status, and a two-way repeated-measures ANOVA is used to determine the effects of age and early pregnancy, along with the peri-ovulatory period, on analyte concentrations. No significant effect of age, ovulation or early pregnancy is detected on the mares’ cortisol, insulin or glucose concentrations; however, there is a significant effect of early pregnancy and ovulation on ACTH concentrations (p = 0.04 and p = 0.04 respectively). ACTH concentrations change around ovulation and with early pregnancy. Therefore, knowledge of a mare’s reproductive status might be beneficial when interpreting ACTH concentrations.

Experimental crossover study on the effects of withholding feed for 24\u2009h on the equine faecal bacterial microbiota in healthy mares

BackgroundAn association between equine gastrointestinal disease causing colic signs and changes in faecal bacterial microbiota has been identified. The reasons for these changes and their clinical relevance has not been investigated. Withholding feed, which is an integral part of managing horses with colic, may contribute to the observed changes in the microbiota and impact interpretation of findings in horses with colic. Study objectives were, therefore, to determine the effect of withholding feed for 24 h on equine faecal bacterial microbiota in healthy mares to differentiate the effects of withholding feed from the changes potentially associated with the disease.ResultsSpecies richness and Shannon diversity (alpha diversity) were significantly lower at the late withheld (10–24 h post withholding feed) and early refed (2–12 h post re-feeding) time points compared to samples from fed horses (P < 0.01). Restoration of species richness and diversity began to occur at the late refed (18–24 h post re-feeding) time points. Horses having feed withheld had a distinct bacterial population compared to fed horses (beta diversity). Bacteroidetes BS11 and Firmicutes Christensenellaceae, Christensenella, and Dehalobacteriaceae were significantly increased in horses withheld from feed primarily during the late withheld and early refed time points. Bacteroidetes Marinilabiaceae and Prevotellaceae, Firmicutes Veillonellaceae, Anaerovibrio, and Bulleidia, and Proteobacteria GMD14H09 were significantly decreased in horses with feed withheld at late withheld, early refed, and late refed time periods (P < 0.01). Changes in commensal gut microbiota were not significant between groups.ConclusionsWithholding feed has a significant effect on faecal bacterial microbiota diversity and composition particularly following at least 10 h of withholding feed and should be taken into consideration when interpreting data on the equine faecal bacterial microbiota in horses.

124 Effect of transvaginal oocyte aspiration on equine blood and peritoneal fluid parameters

Transvaginal aspiration of oocytes (TVA) in the equine industry has gained more relevance and become a valuable technique to produce offspring from subfertile mares. TVA is a semi-invasive procedure and requires handling the ovaries transrectally to position them closely to an ultrasound probe located in the mare’s vagina. Once the ovary lies in close apposition to the ultrasound probe, a 12G needle is inserted through the needle guide, puncturing, aspirating, and scraping each follicle to recover the oocyte. Potential complications described include rectal tears, puncturing of blood vessels, ovarian abscesses, and peritonitis. Occasionally, problems occur after uneventful procedures, such as colic, peritonitis, pain, and anorexia. However, the source of these complications is not fully known. We hypothesize that blood and peritoneal fluid parameters would differ pre- and post-TVA in mares. A few reports provide some parameters after TVA (e.g. peritoneal protein, neutrophils, nucleated cells) without reference to pre-TVA values. These studies have not identified an effect in peritoneal fluid variables due to multiple abdominocenteses. Therefore, our aim was to analyse blood and peritoneal fluid in mares pre- and post-TVA, and to identify changes in parameters of the procedure (duration, number of pokes, number of follicles) and the mares’ clinical responses. Ten healthy mares were selected to undergo the procedure. Thirty minutes before starting TVA, a blood sample was drawn for complete blood count (CBC) and blood chemistry, and abdominocentesis was performed to obtain abdominal fluid and assess the cytology. This same protocol was repeated 24 hours after TVA. Physical exams were performed pre- and post-TVA. Paired t-tests were used to identify differences between groups (pre- and post-TVA). Spearman correlations (ρ) were used to assess the relationship between variables. There was a significant increase in peritoneal lactate (5.65-fold), peritoneal total protein (2.4-fold), and total nucleated cells (46-fold) between pre- and post-samples. These parameters were not associated with operator, number of times the needle was introduced into the ovaries, or number of aspirated follicles. The remaining parameters evaluated in CBC and blood chemistry did not differ. A positive correlation between total peritoneal protein and blood albumin was found post-TVA (ρ=0.72, P=0.01) but not pre-TVA (ρ=−0.1, P=0.65), suggesting an increase in protein level due to bleeding. Clinically, 9 mares were healthy throughout the study except one that presented signs of pain (facial grimace, anorexia, hyperthermia) the day following TVA. In conclusion, we showed changes in the peritoneal fluid during uneventful TVA procedures. The information provided by this research gives further insight into changes potentially caused by a TVA in abdominal fluid parameters. Further studies are necessary to determine expected standards and the duration of the changes after TVA.

Renal failure in a pregnant mare

Renal failure in a pregnant mare

Recovery of Equine Oocytes in Ambulatory Practice and Potential Complications

Field collection of oocytes in mares using transvaginal follicular aspiration (TVA) for embryo production has the potential to revolutionate the equine industry. Protocols for TVA in specialized laboratory settings have been described in the scientific literature since the early 1980s. The objective of this study was to determine the success rate of TVA oocytes recovery under ambulatory conditions. A secondary goal of this study was to determine if TVA is associated with any health complications when performed by recently trained practitioners in the field. Follicles (n = 296) from 66 adult clinically healthy mares were aspirated over a period of 6 days. TVAs were performed by 22 veterinarians with 5–20 years of experience in equine and bovine reproductive medicine, but no previous experience in TVA. Oocytes (n = 145) were recovered. No short- or long-term systemic or local complications were observed following TVA in any of the mares used in this study. Fifty-six out of 66 mares became pregnant within 3 months following TVA. This study shows that with proper training, TVA can be successfully used to obtain equine oocytes with no health complications under field conditions in nonspecialized laboratory settings.

Serial measurements of Paraoxonase-1 (PON-1) activity in horses with experimentally induced endotoxemia

BackgroundParaoxonase-1 (PON-1) is an antioxidant enzyme, whose activity decreases during the acute phase response in many species. Little is known about PON-1 and its role as a negative acute phase protein during septic inflammation in horses, but promising findings about its utility in diagnosing SIRS and predicting the outcome in diseased horses, were recently highlighted. The objective of the study was to investigate the behaviour of PON-1 in horses after experimentally induced endotoxemia. To this aim, PON-1 activity was measured on 66 plasma samples collected from six clinically healthy mares, previously included in another study, before and at multiple time points between 12 and 240 h after intravenous infusion of Escherichia coli O55:B5 lipopolysaccharide (LPS).ResultsCompared with baseline values, a progressive transient decrease of PON-1 activity was observed starting from 24 h post-infusion, with lowest values observed between 3 to 7 days post-infusion, followed by a normalisation to pre-infusion levels the tenth day.ConclusionsThe results of this study suggest that measurement and monitoring of PON-1 activity might be useful to evaluate progression and recovery from endotoxemia in horses. Further studies in horses with naturally occurring sepsis are warranted.

Vaginal Microbiota Is Stable throughout the Estrous Cycle in Arabian Maress.

Lactic acid bacteria (LAB) dominate human vaginal microbiota and inhibit pathogen proliferation. In other mammals, LAB do not dominate vaginal microbiota, however shifts of dominant microorganisms occur during ovarian cycle. The study objectives were to characterize equine vaginal microbiota in mares by culture-dependent and independent methods and to describe its variation in estrus and diestrus. Vaginal swabs from 8 healthy adult Arabian mares were obtained in estrus and diestrus. For culture-dependent processing, bacteria were isolated on Columbia blood agar (BA) and Man Rogosa Sharpe (MRS) agar. LAB comprised only 2% of total bacterial isolates and were not related to ovarian phases. For culture-independent processing, V3/V4 variable regions of the 16S ribosomal RNA gene were amplified and sequenced using Illumina Miseq. The diversity and composition of the vaginal microbiota did not change during the estrous cycle. Core equine vaginal microbiome consisted of Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria at the phylum level. At the genus level it was defined by Porphyromonas, Campylobacter, Arcanobacterium, Corynebacterium, Streptococcus, Fusobacterium, uncultured Kiritimatiaellae and Akkermansia. Lactobacillus comprised only 0.18% of the taxonomic composition in estrus and 0.37% in diestrus. No differences in the relative abundance of the most abundant phylum or genera were observed between estrus and diestrus samples.

Changes in Salivary Analytes of Horses Due to Circadian Rhythm and Season: A Pilot Study.

Simple SummaryThe use of salivary biomarkers is gaining interest in the veterinary field, since saliva is usually easy to obtain and its collection from animals causes less stress than blood sampling. However, our knowledge of the possible factors related to daily and seasonal variations in salivary biomarkers is still in its infancy. In our study, the possible circadian or circannual variations in a panel of salivary biomarkers in horse saliva were evaluated. The results showed that daily and/or seasonal variations can be observed in cortisol, salivary alpha-amylase, total esterase, butyrylcholinesterase, adenosine deaminase, and creatine kinase. Therefore, these factors should be considered for the interpretation of these analytes when measured in horse saliva.This study aims to evaluate the circadian and circannual variations in a panel of analytes in horse saliva that have been previously described as biomarkers related to stress and disease, in order to interpret them correctly when they are measured in this species. This panel of analytes integrated cortisol, salivary alpha-amylase (sAA), lipase (Lip), total esterase (TEA), butyrylcholinesterase (BChE), adenosine deaminase (ADA), γ-glutamyl transferase (gGT), creatine kinase (CK), urea, total bilirubin, total protein (TP), and phosphorus. These analytes were measured in saliva obtained from a population of five clinically healthy mares from 06:30 to 20:30, every 2 h over two consecutive days in two different photoperiod seasons, winter and spring. The temperature and relative humidity did not change between the two consecutive days sampled in each sampled season, and no thermal discomfort was observed. Changes throughout the course of the day were observed for cortisol, sAA, TEA, BChE, ADA, and CK. However, a circadian pattern was only observed for cortisol, TEA, BChE, ADA, and CK. Moreover, the values obtained for sAA, Lip, and BChE were significantly different between seasons, with different daily rhythms for cortisol, TEA, BChE, and ADA depending on the season. In conclusion, this pilot study indicates that the time of the day and the season influence salivary analytes in horses, showing a rhythmic pattern for cortisol, TEA, BChE, ADA, and CK. These factors should thus be taken into consideration for the interpretation of analytes in horse saliva.

History of development tissue therapy and modern prespectives of it’s application in veterinary medicine

Medical products, made of plants, various tissue substrates of animals and humans, have been used for a long time in human and veterinary medicine. Tissue medications of animal origin have been widely used in veterinary science both for prophylactic and therapeutic purposes and as growth factors for fattening of all species. The development of modern biology and some fields of it, human and veterinary medicine in particular, based on the achievements of advanced technologies, provides an opportunity for the appearance of new effective medications in clinical practice. Over time, after the reorganization of the agricultural sector and veterinary medicine as one of its main divisions, tissue medications began to disappear from the range of medical preparations, and they have been almost forgotten and basically not used in recent years. Veterinary medicines market has become overwhelmed with mainly imported synthetic medications and antibiotics. Meanwhile, clinical practice is experiencing an intensive introduction of medications, produced by private factories, which are combinations and mixtures of various existing pharmacological substances. Their annotations indicate a universal and beneficial effect on the body of almost all domestic animals, birds and bees. The paper is aimed at the feasibility study and the restoration necessity of production and use of tissue preparations in veterinary clinical practice. The materials used for the research are the reports published in scientific journals, collections of scientific editions of universities and research establishments, textbooks, newspapers, etc. We have become the first to create tissue medications such as fetoplacentat, made from the uterus and its contents of different pregnancy term of clinically healthy cows, pigs, mares, dogs and cats, and truthenate, made from the larvae of drones aged 5–7 days, for their application in eterinary medicine, obstetrics and gynecology in particular. Production trials of medications carried out on different pregnant animals, have made it possible to identify their corrective effect on the course of pregnancy, childbirth and postpartum period, high preventive activity in the development of functional disorders, subinvolution and inflammatory processes of the genital organs, as well as in the treatment of different forms of infertility in animals and for the improvement of spermatogenesis of male breeders. Further research will focus on the development and implementation of effective methods and measures aimed at elimination of infertility and improvement of the reproduction of farming animals on condition of the use of tissue preparations.

Time to Peak Concentration of Amikacin in the Antebrachiocarpal Joint Following Cephalic Intravenous Regional Limb Perfusion in Standing Horses

The aim of this study was to determine the time (Tmax) to the maximum concentration (Cmax) of amikacin sulphate in synovial fluid of the radiocarpal joint (RCJ) following cephalic intravenous regional limb perfusion (IVRLP) using 2 g of amikacin sulphate. Cephalic IVRLP was performed with 2 g of amikacin sulphate diluted in 0.9% NaCl to a total volume of 100 mL in six healthy adult mixed breed mares. An Esmarch's rubber tourniquet was applied for 30 minutes and the antibiotic solution was infused through a 23-gauge butterfly catheter. Synovial fluid was collected from the RCJ prior to the infusion and at 5, 10, 15, 20, 25 and 30 minutes after completion of IVRLP. The tourniquet was removed after the last arthrocentesis. Synovial fluid amikacin sulphate concentrations were determined by liquid chromatography/tandem mass spectrometry. The calculated mean Tmax occurred at 15 minutes (range: 10-20 minutes) post-perfusion. The highest synovial fluid amikacin sulphate concentration was noted at 10 minutes in 2 horses, 15 minutes in 2 horses and 20 minutes in 2 horses. The highest mean concentration was 1023 µg/mL and was noted at 20 minutes. Synovial mean concentrations were significantly different between 15 and 30 minutes. In this study no Tmax occurred after 20 minutes; thus, 30 minutes of tourniquet application time appear to be excessive. The 20 minutes duration of tourniquet application appears sufficient for the treatment of the RCJ in cephalic IVRLP using 2 g amikacin sulphate in a total volume of 100 mL.