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The objectives of this study were to assess the cooling and freezing of donkey epididymal semen harvested immediately after castration (Experiment 1, n = 4) or after the shipment (24 or 48 h) of epididymides attached to testicles (Experiment 2, n = 14) or dissected apart (Experiment 3, n = 36). In each experiment, semen was frozen immediately (Non-Centrif) in an egg yolk-based semen extender (EY) or after processing through cushion-centrifugation (Centrif) while extended in a skim milk-based extender (SC). In all three experiments, cooled, pre-freeze, and post-thaw epididymal semen was assessed for total motility (TM), progressive motility (PM), plasma membrane integrity (PMI), and high mitochondrial membrane potential (HMMP). Data were analyzed with R using mixed models and Tukey's test as posthoc. Results showed that the cooling of epididymal semen up to 24 h after harvesting did not affect motility parameters or plasma membrane integrity; furthermore, in Experiment 3, the post-thaw evaluation of both Centrif and Non-Centrif achieved similar TM and PM. Collectively, the post-thaw results revealed low motility parameters across groups; while, the PMI and HMMP did not reflect this trend, and the values remained high, suggesting that there was a lack of epididymal sperm activation with either centrifugation or extenders. In summary, freshly harvested and cooled-shipped and cooled semen had satisfactory semen parameters. Future studies need to address donkey epididymal semen fertility in mares and jennies.

Disorders of sex development (DSD) and reproduction are not uncommon among horses, though knowledge about their molecular causes is sparse. Here we characterized a ~200 kb homozygous deletion in chromosome 29 at 29.7-29.9 Mb. The region contains AKR1C genes which function as ketosteroid reductases in steroid hormone biosynthesis, including androgens and estrogens. Mutations in AKR1C genes are associated with human DSDs. Deletion boundaries, sequence properties and gene content were studied by PCR and whole genome sequencing of select deletion homozygotes and control animals. Deletion analysis by PCR in 940 horses, including 622 with DSDs and reproductive problems and 318 phenotypically normal controls, detected 67 deletion homozygotes of which 79% were developmentally or reproductively abnormal. Altogether, 8-9% of all abnormal horses were homozygous for the deletion, with the highest incidence (9.4%) among cryptorchids. The deletion was found in ~4% of our phenotypically normal cohort, ~1% of global warmblood horses and ponies, and ~7% of draught breeds of general horse population as retrieved from published data. Based on the abnormal phenotype of the carriers, the functionally relevant gene content, and the low incidence in general population, we consider the deletion in chromosome 29 as a risk factor for equine DSDs and reproductive disorders.

To examine feasibility and acceptability of providing stepped collaborative care case management targeting posttraumatic stress disorder (PTSD) and pain symptoms after major traumatic injury. Participants were major trauma survivors in Victoria, Australia, at risk of persistent pain or PTSD with high baseline symptoms. Participants were block-randomized, stratified by compensation-status, to the usual care (n = 15) or intervention (n = 17) group (46% of eligible patients). The intervention was adapted from existing stepped collaborative care interventions with input from interdisciplinary experts and people with lived experience in trauma and disability. The proactive case management intervention targeted PTSD and pain management for 6-months using motivational interviewing, cognitive behavioral therapy strategies, and collaborative care. Qualitative interviews explored intervention acceptability. Intervention participants received a median of 7 h case manager contact and reported that they valued the supportive and non-judgmental listening, and timely access to effective strategies, resources, and treatments post-injury from the case manager. Participants reported few disadvantages from participation, and positive impacts on symptoms and recovery outcomes consistent with the reduction in PTSD and pain symptoms measured at 1-, 3- and 6-months. Stepped collaborative care was low-cost, feasible, and acceptable to people at risk of PTSD or pain after major trauma. After hospitalization for injury, people can experience difficulty accessing timely support to manage posttraumatic stress, pain and other concerns. Stepped case management-based interventions that provide individualized support and collaborative care have reduced posttraumatic stress symptom severity for patients admitted to American trauma centers. We showed that this model of care could be adapted to target pain and mental health in the trauma system in Victoria, Australia. The intervention was low cost, acceptable and highly valued by most participants who perceived that it helped them use strategies to better manage post-traumatic symptoms, and to access clinicians and treatments relevant to their needs. [ABSTRACT FROM AUTHOR]

Background: Sinusitis is a common disease of horses yet there are a limited number of reports in the literature that describe the prevalence of infraorbital canal (IOC) pathology and headshaking behaviour in horses diagnosed specifically with primary sinusitis and secondary dental sinusitis. Given the impact that these behaviours can have on horses' intended athletic use, investigation is warranted. Objectives: To determine the occurrence of IOC pathology in horses with concurrent primary or secondary dental sinusitis based on computed tomography (CT) findings and to assess whether the frequency of headshaking behaviour is influenced by the presence of IOC pathology. Study design: Retrospective case series. Methods: Computed tomography studies were assessed for sinusitis (unilateral or bilateral), IOC pathology (unilateral or bilateral) and description of IOC disease including displacement, deformation, periosteal proliferation, hyperostosis, osteolysis and infraorbital nerve exposure. Behaviour outcome was determined by client questionnaire five or more years following CT scan. Results: A total 65 out of 66 horses diagnosed with primary or secondary dental sinusitis demonstrated IOC changes on CT. Hyperostosis (86%), periosteal proliferation (85%) and osteolysis (86%) were common CT findings. Hyperostosis was frequently found to involve both the IOC and supporting bone structure. Five cases were euthanized immediately after CT acquisition or during hospitalisation following diagnostic investigations. Follow‐up was obtained in 48/61 cases, with five horses showing headshaking behaviour. Main limitations: Infraorbital nerve histopathology was not performed. The limited number of cases with no IOC pathology prevented direct comparison between sinusitis groups both with IOC pathology and without IOC changes. The client questionnaire carries a memory bias. Conclusion: Computed tomography changes involving the IOC may not predict headshaking behaviours in sinusitis secondary to dental disease. This finding is important in the context that these behaviours render some horses unusable and unsafe for their intended riding discipline. [ABSTRACT FROM AUTHOR]

Antibiotics are commonly given to horses to treat known bacterial infections or as prophylaxis against the development of infection. Diarrhea is a recognized adverse effect of antimicrobial administration in horses (antimicrobial associated diarrhea, AAD). In veterinary referral centers, AAD has a reported incidence of 22 to 94% (McGorum, 2010), while horses with AAD are 4.5 times more likely to die compared with horses with other types of colitis (Woods and Cohen, 1999). Although all antibiotics have potential to induce diarrhea, some have increased risk due to low oral absorption, biliary excretion, or enterohepatic recycling. The role of dysbiosis in the gut microbiota is a key feature in the pathogenesis of AAD. Disruption of commensal bacteria of the hindgut allows for pathogen overgrowth and alteration in microbial metabolic function. To date, 16S rRNA sequencing has been used to characterize the effects of penicillin, trimethoprim sulfa, doxycycline, erythromycin, ceftiofur, enrofloxacin, oxytetracycline and metronidazole on the fecal microbiome in healthy horses (Costa et al., 2015; Arnold et al., 2020; Liepman, 2022). Regardless of mechanism of action, antibiotics typically reduce diversity (species richness and evenness) and alter the bacterial community structure of the fecal microbiome, even when animals maintain normal health status and fecal character (Arnold et al., 2021). These effects typically require at least 30 days before the microbiome returns to baseline (Gomez et al, 2023). In horses with colitis, changes in the microbiome are often significant. Horses with AAD demonstrate the most severe disruption of the fecal microbiome compared with horses with Salmonella or undifferentiated colitis. These changes are evident from the phylum to species level, and include important phyla such as Bacteroidetes, Firmicutes, Fibrobacter and Verrucomicrobia (Arnold et al., 2020; Liepman, 2022; Costa et al., 2015). Alterations in the microbiota result in functional metabolic changes that are clinically manifested by diarrhea. Potential changes include alterations in the metabolism of bile acids, carbohydrates including short chain fatty acids, amino acids, lipids and proteins. There is also evidence that antimicrobials increase intestinal permeability via effects on tight junction proteins and by compromising the barrier function provided by the mucus layer between the enterocytes and gut lumen. Recent work comparing horses on antimicrobials that did and did not develop diarrhea confirm that horses with diarrhea have depletion of Verrocomicrobia and metabolites related to gastrointestinal barrier function (Arnold et al., 2021). [ABSTRACT FROM AUTHOR]

The non-nitrogenous bisphosphonate clodronate disodium (CD) inhibits bone resorption and is approved for horses ≥ 4 yr of age. Extra-label use in juvenile exercising horses is a concern due to potential interference with normal bone growth and development. The objective was to determine the effects of single and repeated doses of CD on bone optical density (radiographic bone aluminum equivalence; RBAE) and lameness score (LS) in juvenile, exercising horses. Quarter Horses [n = 32 (16 geldings, 16 fillies); 500 ± 13 d of age; body weight (BW) = 336 ± 26 kg]) were used to test the hypothesis that horses receiving CD would have greater RBAE and reduced LS, and additional doses of CD would accentuate these effects. In a 168-d trial, horses were housed in individual stalls, and fed 1% BW/d concentrate and ad libitum Coastal bermudagrass hay. Horses were exercised 5 d/wk following a phase-based progressive workload using a free stall exerciser; Phase I (d 0 to 84) mimicked sales preparation whereas Phase II (d 85 to 168) simulated early training. Horses were stratified by age, BW, sex, and initial RBAE into control (CON; n = 8), single dose CD (1X; d 84; n = 8), 2-doses CD (2X; d 0 and d 84; n = 8), and four-doses CD (4X; d 0, d 42, d 84, d 126; n = 8). On injection d (0, 42, 84, and 126), horses were administered either 1.8 mg/kg BW CD (OSPHOS) or isovolumetric saline. On d 0, 84, and 168, dorsalpalmar and lateral-medial radiographs of the left third metacarpus and metatarsus were captured. An aluminum step wedge was included in each image and raw images were analyzed (Quantity One Basic, BioRad) to determine total bone RBAE at 1 cm distal to the nutrient foramen. On d 0 and d 168, horses were evaluated by a veterinarian unaware of treatment group and assigned a LS according to the American Association of Equine Practitioners Lameness Scale. The RBAE and LS data were analyzed via PROC MIXED and PROC GENMOD, respectively, in SAS. For each view, RBAE increased from d 0 to d 84 and remained increased to d 168 (P ≤ 0.02), regardless of treatment, likely due to progressing exercise protocol. On d 0, 29 horses were LS 0, 1 horse was LS 1, and 2 horses were LS 2; LS did not differ among treatment groups (P = 0.61). On d 168, 5 horses were LS 0, 13 horses were LS 2, 11 horses were LS 3, and 1 horse was LS 4; LS did not differ among treatments (P = 0.16). Based on these results, the hypothesis that single or repeated CD administration would increase RBAE and decrease lameness in juvenile, exercised Quarter Horses was rejected. [ABSTRACT FROM AUTHOR]

Intra-articular corticosteroids (IAC) are administered to horses experiencing joint inflammation, at times with a concurrent articular antibiotic for prophylaxis. However, information regarding the biologic effects of IAC and potential for microbial contamination in IAC-administered joints is limited in young, exercising horses. The objective of this study was to evaluate the impact of IAC on traditional microbial culture, synovial fluid (SF) volume, and the presence of IAC; hypothesizing negative microbial culture, no change of SF volume, and detection of IAC in SF post-administration. To test this, 2-yr-old Quarter Horses {n = 24; body weight (BW) = 409 ± 6 kg; 825 ± 21 d of age; 12 fillies, 12 geldings) were stratified by BW, age, and sex, and randomly assigned to 1 of 3 treatment groups for a 56-d trial. Horses were housed individually in stalls with runs and fed concentrate offered at 0.75% BW/d split evenly every 12 h with ad libitum Coastal bermudagrass hay and water. Every 21 d, BW and body condition score (BCS) were recorded, and concentrate was adjusted accordingly. Horses were exercised 5 d/wk, up to 45 min/d in a progressive workload using a free-stall exerciser. Blood was collected postexercise during wk 1, 4, and 7 via jugular venipuncture and lactate (BL) was measured to monitor increasing workload. Intra-articular treatments consisted of 0 mg (CON; n = 8), 6 mg (CORT6; n = 8), or 12 mg (CORT12; n = 8) of triamcinolone acetonide (TA). On d 0, treatments were administered in one randomly selected radiocarpal joint per horse. Following aseptic preparation, SF from both radiocarpal joints was collected before administration (pre), and on d 7, 14, 28, and 56. Volume of SF was recorded, and standard microbial culture and detection of TA were performed by a commercial laboratory. Data were analyzed using PROC MIXED (BW, BCS, BL, SF volume) and PROC GLM (TA) of SAS. For SF volume, data from the contralateral radiocarpal joint were used as a covariate. Horses increased BW and BCS over time (P ≤ 0.01) across treatments. Post-exercise BL increased from wk 1 to 4 and remained increased through wk 7 (P < 0.01). Microbial culture for all horses and time points were negative. Concentration of TA was not evaluated on d 7 due to limited SF volume, but TA was detected on d 14 and concentration did not differ between CORT6 and CORT12 (P = 0.13). Recorded SF volume did not change from d 0 (pre) to 7 but increased to d 56, regardless of treatment (P < 0.01). These results indicate that, based on standard culture, TA administration did not result in positive microbial culture, or decrease SF volume in young horses undergoing regular exercise, however, TA was detectable to d 14. [ABSTRACT FROM AUTHOR]

Introduction: Persistent high rates of prescription opioid use and harms remain a concern in Australia, Europe and North America. Research priority setting can inform the research agenda, strategic responses and evidence‐based interventions. The objective of this study was to establish research priorities related to the safe and effective use of prescription opioids in general practice. Methods: Consumers, clinicians and policy makers were invited to participate in a structured consensus workshop in May 2021. A modified nominal group technique was used to explore research priorities for the safe and effective use of opioids in Australian general practice. Research priorities were identified, consolidated and prioritised using a structured process. Results: Seventeen consumer, medical, pharmacy, nursing, allied health and policy participants generated 26 consolidated priorities across three domains: (i) consumer‐focused priorities; (ii) clinician and practice‐focused priorities; and (iii) system and policy‐focused priorities. The highest ranked research priorities in each of the domains were consumer characteristics that influence opioid prescribing and outcomes, opioid deprescribing strategies, and system‐level barriers to prescribing alternatives to opioids, in the consumer, clinician and practice, and system and policy domains, respectively. Discussion and Conclusion: The priorities reflect opportunities for research priority setting within Australian general practice. The priorities provide a map for future qualitative and quantitative research that will inform safe and effective opioid prescribing. [ABSTRACT FROM AUTHOR]

Concerns over the extra-label use of bisphosphonates in skeletally immature horses has extended to include its likely re-release from bone into circulation in response to stressors. Therefore, the objective of this study was to be first to quantify the re-release of clodronate in response to controlled stressors. Yearling Quarter horses (n = 32) were stratified by age, body weight (BW), sex, and initial bone optical density into four treatment groups for a 168-d trial. Treatments consisted of control (CON; n = 8), single-dose (1X; d 84; n = 8), 2-dose (2X; d 0 and 84; n = 8), and four-dose groups (4X; d 0, 42, 84, and 126; n = 8). All horses received iso-volumetric intramuscular injections of either 1.8 mg/kg BW clodronate disodium (OSPHOSÒ) or saline (placebo) on d 0, 42, 84, and 126. Horses were housed in individual stalls (3.6 m × 7.2 m). Diets were formulated to meet nutrient requirements including concentrate offered every 12-h and ad libitum coastal bermudagrass (Cynodon dactylon) hay and water. Horses were exercised 5 d/ wk with a progressive workload, and maximum exercise intensity was reached on d 120, verified via heart rate and blood lactate. On d 120, blood samples were collected prior to the start of the exercise bout (pre), and 0-, 0.5-, 1-, 3-, 12-, and 24-h post exercise. Both tuber coxae of each horse were biopsied on d 168 with blood samples collected prior to (pre), and 0, 12, 24, 48, 72, 168, and 336 h post biopsy. Clodronate was quantified in plasma from exercise and biopsy samples using liquid chromatography tandem mass spectrometry. Exercise and biopsy serum were analyzed for cortisol, a marker of stress, while biopsy serum was analyzed for substance P, a pain marker. Data were analyzed using SAS PROC MIXED. Exercise did not result in clodronate re-release as there was no treatment x time interaction or time effect (P ≥ 0.44), but clodronate was dose-dependently greater in treated groups throughout the exercise collection period (P < 0.01). A treatment x time interaction (P ≤ 0.01) was observed for clodronate concentrations surrounding bone biopsy in which concentrations decreased at 168 h post biopsy before increasing at 336 h in 4X horses. Substance P increased over time (P < 0.01) following biopsy, regardless of treatment. Despite decreasing over time (P ≤ 0.01), there were similarly no treatment differences (P ≥ 0.46) nor treatment ´ time interactions (P ≥ 0.78) for cortisol following either exercise or bone insult. In conclusion, submaximal exercise did not result in re-release of clodronate; however, direct bone insult disrupted clodronate concentrations in horses receiving 4 administrations of bisphosphonate despite no treatment impact on stress or pain indicators. [ABSTRACT FROM AUTHOR]

Intra-articular lipopolysaccharide (LPS) is an established model for inducing robust, transient inflammation and increasing cartilage metabolism for up to 24 h following administration. Currently, there is limited information evaluating the long-term effects of this gram-negative endotoxin on the intra-articular environment of young horses. Therefore, the objective of this study was to determine the lasting effects of a single administration of intra-articular LPS on synovial biomarkers of inflammation and cartilage metabolism over 35 wk. Quarter Horse yearlings [n = 10; body weight (BW) = 409 ± 24 kg; 619 ± 16 d of age; fillies, (n = 7) and geldings (n = 3), originating from a single herd were used to test the hypothesis that intraarticular inflammation and cartilage metabolism would not be elevated at 35 wk post-injection. Each horse had one randomly selected radiocarpal joint injected with 0.8 mL of a 0.5 ng LPS solution derived from Escherichia coli O55:B5 (INFL; Sigma-Aldrich, St. Louis, MO) and the contralateral joint received sterile lactated Ringer’s solution (CON) as a control. Synovial fluid was obtained on 0 (before injection of LPS), 2, and 35 wk following arthrocentesis. Synovial fluid samples were analyzed in duplicate for carboxypropeptide of type II collagen (CPII), collagenase cleavage neopeptide (C2C), chondroitin sulfate 846 epitope (CS846), and prostaglandin E2 (PGE2 ) by commercial ELISA (IBEX Pharm, Montreal, Quebec, CA and Arbor Assays, Ann Arbor, MI). Biomarkers were analyzed using PROC MIXED of SAS v9.4 with repeated measures (time). The model included treatment (CON, INFL), time (week), and treatment × time interaction as fixed effects. Synovial biomarkers PGE2, CPII, and CS846 changed over time (P < 0.01) regardless of LPS administration, increasing from 0 to 2 wk and declining to below baseline concentrations at wk 35. There was a treatment × time interaction (P ≤ 0.01) in which INFL joints had decreased catabolic C2C concentrations compared with CON joints at wk 35. Therefore, LPS administration did not increase cartilage metabolism and inflammation levels at 2 or 35 wk post-LPS induction, indicating no negative effects compared with the contralateral controls. This study supports using intra-articular LPS in young horses without negative long-term effects. [ABSTRACT FROM AUTHOR]