Characterization of cannabinoid plasma concentration, maternal health, and cytokine levels in a rat model of prenatal Cannabis smoke exposure.

Cannabis sativa has gained popularity as a "natural substance", leading many to falsely assume that it is not harmful. This assumption has been documented amongst pregnant mothers, many of whom consider Cannabis use during pregnancy as benign. The purpose of this study was to validate a Cannabis smoke exposure model in pregnant rats by determining the plasma levels of cannabinoids and associated metabolites in the dams after exposure to either Cannabis smoke or injected cannabinoids. Maternal and fetal cytokine and chemokine profiles were also assessed after exposure. Pregnant Sprague-Dawley rats were treated daily from gestational day 6-20 with either room air, i.p. vehicle, inhaled high-Δ9-tetrahydrocannabinol (THC) (18% THC, 0.1% cannabidiol [CBD]) smoke, inhaled high-CBD (0.7% THC, 13% CBD) smoke, 3 mg/kg i.p. THC, or 10 mg/kg i.p. CBD. Our data reveal that THC and CBD, but not their metabolites, accumulate in maternal plasma after repeated exposures. Injection of THC or CBD was associated with fewer offspring and increased uterine reabsorption events. For cytokines and chemokines, injection of THC or CBD up-regulated several pro-inflammatory cytokines compared to control or high-THC smoke or high-CBD smoke in placental and fetal brain tissue, whereas smoke exposure was generally associated with reduced cytokine and chemokine concentrations in placental and fetal brain tissue compared to controls. These results support existing, but limited, knowledge on how different routes of administration contribute to inconsistent manifestations of cannabinoid-mediated effects on pregnancy. Smoked Cannabis is still the most common means of human consumption, and more preclinical investigation is needed to determine the effects of smoke inhalation on developmental and behavioural trajectories.

Prolonged holding time and sampling protocol affects viscoelastic coagulation parameters as measured by the VCM-Vet™ using fresh equine native whole blood.

OBJECTIVES: Determine the effect of sample holding time and single sample reuse on viscoelastic coagulation parameters when using fresh equine native whole blood. ANIMALS: 8 healthy adult horses from a university teaching herd. PROCEDURES: Blood collected by direct jugular venipuncture (18 ga needle, 3 mL syringe) was held at 37 °C for 2, 4, 6, or 8 minutes according to 1 of 2 protocols. Syringes were gently inverted twice, a small amount of blood was expressed, testing cartridges were filled, and placed within the VCM-Vet™ device (Entegrion Inc). Protocol A: samples were processed from a single syringe. Protocol B: 4 syringes were drawn through a single needle. VCM-Vet™ measures assessed included clot time (CT), clot formation time (CFT), alpha angle (AA), amplitude at 10/20 minutes (A10/A20), maximal clot firmness (MCF), and lysis index at 30/45 minutes (LI30/LI45). Differences over time were examined using the Friedman test and post hoc Wilcoxon Rank Sum Test with Bonferroni correction, P ≤ .05. RESULTS: Following Protocol A, there was a significant effect of holding time for CT (P = .02), CFT (P = .04), and AA (P = .05). CT and AA decreased over time, while CFT increased. Samples handled by Protocol B showed no significant difference over time for any of the VCM-Vet™ parameters. CLINICAL RELEVANCE: Sample holding time and handling protocol impact VCM-Vet™ testing results of fresh equine native whole blood. Viscoelastic coagulation samples tested using the VCM-Vet™ may be held unagitated for up to 8 minutes after collection while warm, but should not be reused.

Hyponatremia in horses with septic pneumopathy.

BACKGROUND: Hyponatremia is common in horses with bacterial pleuropneumonia, but no further characterization of this abnormality has been reported. OBJECTIVES: Describe admission plasma sodium concentration ([Na]) in horses with septic pneumopathy and evaluate any association of plasma [Na] with markers of systemic inflammation. ANIMALS: Medical records of horses >1 month of age that between 2008 and 2021 had a transtracheal aspirate (TTA) performed, abnormal TTA cytology, positive TTA culture, pulmonary disease on ultrasonography, radiography or both, and plasma [Na] assessed by direct ion-selective-electrode (dISE). Horses with concurrent diarrhea or azotemia were excluded. METHODS: Clinical and clinicopathological variables of interest between hypo- and normonatremic horses were compared. Spearman correlation and Fisher exact tests were used to identify significant associations (P < .05). RESULTS: Twenty of 35 horses had hyponatremia (median, 132 mmol/L; 25-75th interquartile range [IQR], 129.7-133.1 mmol/L; reference range, 134.2-138.4 mmol/L). A higher proportion of horses with systemic inflammatory response syndrome (SIRS) had hyponatremia (P = .01). Hyponatremic patients had higher mean plasma fibrinogen concentration (461 ± 160.5 mg/dL; P = .01) and higher rectal temperature (38.8 ± 0.7°C; P = .02) than normonatremic horses. Negative correlations were found between plasma [Na] and fibrinogen (P = .001; ρ = -0.57) concentrations and between plasma [Na] and rectal temperature (P = .001; ρ = -0.51). Presence or absence of pleural effusion did not influence severity of hyponatremia. Mean duration of hospitalization was longer (P = .04) in hyponatremic horses (9.8 ± 6.6 days). CONCLUSIONS AND CLINICAL IMPORTANCE: Hyponatremia at admission is associated with the presence of inflammation, SIRS, and with longer duration of hospitalization.

Plasma ascorbic acid, antioxidant capacity, and reactive oxygen species in healthy foals.

OBJECTIVE: To describe ascorbic acid (AA) concentrations, plasma antioxidant capacity (PAC) and markers of oxidative stress, as measured by derivatives of reactive oxygen metabolites (dROMs), in healthy foals at birth and during the first month of life. SAMPLES: Venous blood samples were collected from healthy Standardbred (n = 13) and Quarter Horse (n = 10) foals. Plasma AA, PAC, and dROMs were assessed at 3 to 12 hours, 3 days, and 1, 2, and 4 weeks of age. PROCEDURES: AA was measured via high-performance liquid chromatography. PAC and dROMs were measured with a free radical analytical system. Comparisons of AA, PAC, and dROMs at different time points were assessed. RESULTS: Mean ± standard deviation AA concentrations at 3 to 12 hours (44.7 ± 19.6 µmol/L; P ≤ .01), 1 (48.6 ± 22.5 µmol/L; P ≤ .001), and 2 weeks (41.8 ± 15.8 µmol/L; P ≤ .001) were higher than at 4 weeks of age (28.5 ± 12.7 µmol/L). Both PAC and dROMs significantly increased at different time points compared to 3 to 12 hours of age. CLINICAL RELEVANCE: Healthy foals have higher plasma AA concentrations shortly after birth, which then gradually decrease throughout the first month of life, suggesting that AA may represent a key antioxidant in the postnatal period. The concurrent increase in PAC and dROMs suggests that dynamic development of oxidative balance occurs after birth in foals. Development of AA, PAC, and dROM reference ranges in healthy foals could be used to guide therapeutic interventions and monitor during disease states characterized by increased oxidative stress.

The effects of acute Cannabis smoke or Δ9-THC injections on the trial-unique, nonmatching-to-location and five-choice serial reaction time tasks in male Long-Evans rats.

Executive functions including working memory (WM) and attention are altered following Cannabis exposure in humans. To test for similar effects in a rodent model, we exposed adult male rats to acute Cannabis smoke before testing them on touchscreen-based tasks that assess these executive processes. The trial-unique, delayed nonmatching-to-location (TUNL) task was used to evaluate WM, task performance at different spatial pattern separations, and response latencies. The five-choice serial reaction time task (5-CSRTT) was used to measure attention, impulsivity, perseveration, and response latencies. Rats were exposed acutely to high- Δ9-tetrahydrocannabinol (THC), low-CBD (Mohawk) and low-THC, high-CBD (Treasure Island) strains of Cannabis smoke using a chamber inhalation system. The effects of Cannabis smoke were directly compared to systemic Δ9-THC injection (3.0 mg/kg; i.p.). TUNL task performance was significantly impaired following acute high-THC smoke exposure or THC injections, but not low-THC smoke exposure, with no effects on response latencies. Fewer total trials and selection trials were also performed following THC injections. Performance was poorer for smaller separation distances in all groups. Neither acute smoke exposure, nor injected THC, impacted attentional processes, impulsivity, perseverations, or response latencies in the 5-CSRTT. Pharmacokinetic analysis of rat plasma revealed significantly higher THC levels following injections than smoke exposure 30 min following treatment. Exposure to low-THC, high-CBD Cannabis smoke significantly increased CBD in plasma, relative to the other treatments. Taken together, our results suggest that WM processes as measured by the TUNL task are more sensitive to THC exposure than the attentional and impulsivity measures assessed using the 5-CSRTT.

Pharmacokinetics and pharmacodynamics of intravenous and oral apixaban in horses.

Large vessel and microvascular thrombi are common complications in systemically ill horses contributing to patient morbidity and mortality. Apixaban, an oral factor Xa inhibitor, shows excellent efficacy against stroke and deep vein thrombosis in humans. The purpose of this study was to determine serum apixaban concentrations and anti-factor Xa activity in horses after orally administered apixaban. Five horses received a single dose of intravenous (0.09 mg/kg) and oral (1 mg/kg) apixaban in a cross-over design. Serum apixaban concentrations and anti-Xa activity were measured serially via liquid chromatography-tandem mass spectrometry and a commercial assay, respectively, for 12 hr following oral administration. Apixaban was detected in all horses after both oral and intravenous administration. Oral administration yielded a mean maximum concentration of 60.3 ng/ml (59.4-111 ng/ml), mean time to maximum concentration of 0.5 hr (0.5-2), mean half-life of 6.2 hr (4.6-8.3), and mean oral bioavailability of 10% (3.8-17.4). After oral administration, anti-Xa activity had a strong positive relationship with serum apixaban and was best represented by a dose-response model with the following parameters: E0  = 5.00 ng/ml, EMAX  = 311 ng/mL, EC50  = 267 ng/ml, and n = 1.58. Anti-Xa activity was significantly higher 2 hr post-administration compared with baseline (p = .032). Despite low oral bioavailability, administration of 1 mg/kg oral apixaban, in healthy horses, achieves serum concentrations similar to those reported in humans. Apixaban has potential clinical utility in horses and warrants further investigation.

Normal regression of the internal umbilical remnant structures in Standardbred foals.

BACKGROUND: Diseases affecting the internal umbilical remnant are common in foals. Ultrasound is used to diagnose abnormalities of these structures, and to monitor treatment. However, little is known about the progression of normal internal umbilical remnant regression. OBJECTIVES: To document normal regression of the umbilical remnant in foals over the first 6 weeks of life. STUDY DESIGN: Prospective, longitudinal cohort study. METHODS: Weekly ultrasound examinations were performed beginning at 1 day of age in 34 healthy Standardbred foals. The umbilical vein was measured in cross section just cranial to the external umbilicus, at the level of the liver and midway between these points. The umbilical arteries were measured in cross section just caudal to the external umbilicus, at the apex of the bladder and at the midpoint of the bladder. The urachus was imaged longitudinally as it exited the bladder. Reduction in size over time was evaluated as percentage change in diameter. RESULTS: Structure diameter decreased linearly over time. The largest measurements were at 24 hours of age with a median umbilical vein diameter of 0.83 cm (IQR 0.77-1.02 cm), median umbilical artery diameter of 0.61 cm (IQR 0.56-0.70 cm) and median urachal diameter of 1.07 cm (IQR 1.02-1.14 cm). There was a significant reduction in diameter for all structures (16.0%-21.9%; corrected P < .001) within the first week of life. MAIN LIMITATIONS: All foals were of a single breed, and examinations and measurements were performed by multiple individuals. CONCLUSIONS: The internal umbilical remnants (umbilical vein/arteries, and urachus) rapidly regress over the first few weeks of life. The median internal umbilical remnant diameters reported here are smaller than previously reported values, emphasising the importance of accounting for age when diagnosing umbilical abnormalities. In a healthy Standardbred foal, normal structures are difficult to definitively identify via transcutaneous ultrasound by 5-6 weeks of age.

Uterine responses and equine chorionic gonadotropin concentrations after two intrauterine infusions with kerosene post early fetal loss in mares.

Pregnancy loss during the normal lifespan of endometrial cups (∼37-120-150 days of gestation) may affect a mare's ability to conceive again in the same breeding season, as equine chorionic gonadotropin (eCG) secretion by retained endometrial cups can lead to abnormal ovulations and follicular growth. While intrauterine kerosene infusion has anecdotally been proposed as a treatment for endometrial cup retention, there are no controlled studies evaluating kerosene's ability to enhance endometrial cup regression following abortion. The objectives of this study were to assess uterine response, systemic side effects, and efficacy of intrauterine kerosene infusions after abortion. We hypothesized that kerosene infusions would hasten regression of endometrial cups without detrimental effects on the endometrium and the mare's general health. Twelve light-breed mares were enrolled in the study after an experimentally induced abortion with cloprostenol (n = 12) by 60 ± 2 days of gestation. Mares were randomly allocated to receive an intrauterine infusion with 500 mL of kerosene (n = 6) or 500 mL saline (n = 6) on days 21 and 35 after pregnancy termination. Uterine biopsies were collected at days 7, 21, 35, and 49 post-abortion to evaluate the degree of endometrial fibrosis with Picrosirius Red Stain and to be graded according to the Kenney & Doig 1986 classification. Furthermore, histomorphometry analysis of the endometrium lining, glandular epithelium and glandular density was performed. Endometrial lymphocyte B CD20+, lymphocyte T CD3+, and macrophage IBA-1+ cell populations were characterized by immunohistochemistry. Physical examinations, blood cell counts, and serum biochemistry were performed before, and for 2 days after each uterine infusion. Serum samples were collected for assessment of eCG concentrations. Continuous data were analyzed with MIXED procedure with repeated measures in SAS, categorical data with LOGISTIC procedure of SAS. Significance was set at p < 0.05. Kerosene infusion did not affect complete blood cell counts, serum chemistry parameters, or physical examinations. Concentrations of eCG decreased over time (p < 0.001), but there were no differences between groups or time by group interactions (p = 0.72). Histological evaluation of the uterus showed no signs of increased fibrosis or degeneration in the treatment group. In conclusion, while kerosene infusions did not appear to have detrimental effects on mare health, our findings suggest that the use of kerosene in the uterus does not enhance the regression of endometrial cups by 49 days post-abortion.

Pulmonary disposition and pharmacokinetics of minocycline in adult horses.

OBJECTIVE To determine pharmacokinetics and pulmonary disposition of minocycline in horses after IV and intragastric administration. ANIMALS 7 healthy adult horses. PROCEDURES For experiment 1 of the study, minocycline was administered IV (2.2 mg/kg) or intragastrically (4 mg/kg) to 6 horses by use of a randomized crossover design. Plasma samples were obtained before and 16 times within 36 hours after minocycline administration. Bronchoalveolar lavage (BAL) was performed 4 times within 24 hours after minocycline administration for collection of pulmonary epithelial lining fluid (PELF) and BAL cells. For experiment 2, minocycline was administered intragastrically (4 mg/kg, q 12 h, for 5 doses) to 6 horses. Plasma samples were obtained before and 20 times within 96 hours after minocycline administration. A BAL was performed 6 times within 72 hours after minocycline administration for collection of PELF samples and BAL cells. RESULTS Mean bioavailability of minocycline was 48% (range, 35% to 75%). At steady state, mean ± SD maximum concentration (Cmax) of minocycline in plasma was 2.3 ± 1.3 µg/mL, and terminal half-life was 11.8 ± 0.5 hours. Median time to Cmax (Tmax) was 1.3 hours (interquartile range [IQR], 1.0 to 1.5 hours). The Cmax and Tmax of minocycline in the PELF were 10.5 ± 12.8 µg/mL and 9.0 hours (IQR, 5.5 to 12.0 hours), respectively. The Cmax and Tmax for BAL cells were 0.24 ± 0.1 µg/mL and 6.0 hours (IQR, 0 to 6.0 hours), respectively. CONCLUSIONS AND CLINICAL RELEVANCE Minocycline was distributed into the PELF and BAL cells of adult horses.