Authentic Conversations about Self-Care with Fourth-Year Veterinary Medical Students.

Expanding literature on well-being within veterinary medicine has been instrumental in recognizing the prevalence of psychological distress among students and practitioners and promoting awareness and advocacy for well-being within teaching institutions, professional associations, and the workplace. However, greater focus on distress has also been critiqued for overemphasizing illness and reactive interventions, and a call has been made for more balanced conversations about veterinary well-being, with focus on strengths and proactive interventions. This Best Practices article highlights a proactive, strengths-focused intervention, aimed at increasing fourth-year students' awareness of self-care while in training and in their transition to the profession. Authentic conversations about self-care is a required part of clinical training at Kansas State University College of Veterinary Medicine. All students attend a private meeting with a behavioral scientist, engaging in an authentic conversation about their experience of stress and coping strategies. Current practices in providing stress management interventions are restricted to pre-clinical training. Authentic conversations about self-care are presented here as an alternative to current practices, which the authors argue are not adequately addressing students' needs during clinical training and the transition to the profession. Potential implications of providing self-care interventions during the clinical year of training include greater student engagement, increasing awareness, and self-efficacy as students make efforts to maintain well-being both in training and in the professional realm.

Assessment and Revision of the Veterinary Internship and Residency Matching Program Standardized Letter of Reference.

The Veterinary Internship and Residency Matching Program (VIRMP) recently revised its electronic standardized letter of reference (SLOR) to improve the quality and usefulness of the data obtained from it and to enhance the relevance of non-cognitive and cognitive candidate attributes assessed. We used a stepwise process including a broad survey of SLOR readers and writers, analysis of past SLORs, and a multi-wave iterative revision that included key stakeholders, such as residency and internship program directors from academia and private practice. Data from the SLOR survey and analysis of past SLOR responses identified opportunities to improve applicant differentiation, mitigate positive bias, and encourage response consistency. The survey and other analytics identified and confirmed performance domains of high relevance. The revised SLOR assesses four performance domains: knowledge base and clinical skills, stress and time management, interpersonal skills, and personal characteristics. Ratings within the revised SLOR are predominantly criterion-referenced to enhance discernment of candidate attributes contained within each domain. Questions assessing areas of strength and targeted mentoring were replaced with free-text boxes, which allow writers to comment on positive and neutral/negative ratings of attributes within domains. Minor revisions were made to certain questions to enhance readability, streamline responses, or address targeted concerns identified in the SLOR survey or stakeholder review. The revised SLOR was deployed in the 2020 VIRMP; data from a survey of writers (n = 647) and readers (n = 378) indicate that the redesign objectives were achieved.

Seroprevalence, frequency of leptospiuria, and associated risk factors in horses in Kansas, Missouri, and Nebraska from 2016-2017.

Leptospirosis is a worldwide veterinary and public health concern, and well recognized infectious disease of horses. Seroprevalence rates vary with geography, but many studies have confirmed a high exposure rate. The correlation between seropositivity and shedding status has not been made in horses, however. The aims of this study were to use semi-nested PCR on urine from apparently healthy horses to determine period prevalence of leptospiral shedding and to correlate these findings with MAT results to establish associations with client based survey data regarding horse management and environment. Serum and free-catch urine were collected from 204 healthy horses between May 2016-December 2017. Serum was used to determine GGT, creatinine concentrations, and six serovar MAT. Urine samples were submitted for PCR testing of leptospiral 23S rRNA. Client consent and survey data were collected for all subjects. Potential risk factors included drinking water source, exposure to livestock and dogs, geographical location, season, and precipitation. Two horses were positive on urine PCR for leptospirosis (shedding prevalence 1%), yet only one had a high reciprocal MAT titer of ≥ 800. Both horses were negative on urine PCR one month later without treatment. Approximately 77% of horses (157/204) were seroreactive (MAT reciprocal titer ≥ 100) with titers to serogroup Australis detected more frequently than others (47.5%; (97/204)). Apparently healthy horses infrequently shed Leptospira spp. in urine, yet seroreactivity in clinically normal horses is high (77%), confirming high exposure rates to Leptospira spp. in the Central Midwest.

Bayesian Geostatistical Analysis and Ecoclimatic Determinants of Corynebacterium pseudotuberculosis Infection among Horses.

Kansas witnessed an unprecedented outbreak in Corynebacterium pseudotuberculosis infection among horses, a disease commonly referred to as pigeon fever during fall 2012. Bayesian geostatistical models were developed to identify key environmental and climatic risk factors associated with C. pseudotuberculosis infection in horses. Positive infection status among horses (cases) was determined by positive test results for characteristic abscess formation, positive bacterial culture on purulent material obtained from a lanced abscess (n = 82), or positive serologic evidence of exposure to organism (≥ 1:512)(n = 11). Horses negative for these tests (n = 172)(controls) were considered free of infection. Information pertaining to horse demographics and stabled location were obtained through review of medical records and/or contact with horse owners via telephone. Covariate information for environmental and climatic determinants were obtained from USDA (soil attributes), USGS (land use/land cover), and NASA MODIS and NASA Prediction of Worldwide Renewable Resources (climate). Candidate covariates were screened using univariate regression models followed by Bayesian geostatistical models with and without covariates. The best performing model indicated a protective effect for higher soil moisture content (OR = 0.53, 95% CrI = 0.25, 0.71), and detrimental effects for higher land surface temperature (≥ 35°C) (OR = 2.81, 95% CrI = 2.21, 3.85) and habitat fragmentation (OR = 1.31, 95% CrI = 1.27, 2.22) for C. pseudotuberculosis infection status in horses, while age, gender and breed had no effect. Preventative and ecoclimatic significance of these findings are discussed.

Aging reduces glial uptake and promotes extracellular accumulation of Aß from a lentiviral vector.

We used a lentiviral system for expressing secreted human Aß in the brains of young and old APOE knock-in mice. This system allowed us to examine Aß metabolism in vivo, and test the effects of both aging and APOE genotype, two of the strongest risk factors for Alzheimer's disease. We injected the Aß1-42 lentivirus into the motor cortex of young (2 month old) and old (20-22 month old) APOE3 and APOE4 mice. After 2 weeks of lentiviral expression, we analyzed the pattern of Aß accumulation, glial activation, and phosphor-tau. In young mice, Aß accumulated mainly within neurons with no evidence of extracellular Aß. Significantly higher levels of intraneuronal Aß were observed in APOE4 mice compared to APOE3 mice. In old mice, APOE4 predisposed again to higher levels of Aß accumulation, but the Aß was mainly in extracellular spaces. In younger mice, we also observed Aß in microglia but not astrocytes. The numbers of microglia containing Aß were significantly higher in APOE3 mice compared to APOE4 mice, and were significantly lower in both genetic backgrounds with aging. The astrocytes in old mice were activated to a greater extent in the brain regions where Aß was introduced, an effect that was again increased by the presence of APOE4. Finally, phospho-tau accumulated in the region of Aß expression, with evidence of extracellular phospho-tau increasing with aging. These data suggest that APOE4 predisposes to less microglial clearance of Aß, leading to more intraneuronal accumulation. In older brains, decreased clearance leads to more extracellular Aß, and more downstream consequences relating to astrocyte activation and phospho-tau accumulation. We conclude that both aging and APOE genotype affect pathways related to Aß metabolism by microglia.

Transient Fanconi syndrome in Quarter horses.

Two Quarter horses with weight loss had glucosuria, euglycemia, and a mild metabolic acidosis suggesting a proximal renal tubular defect. Further testing revealed transient generalized aminoaciduria, lactic aciduria, and glucosuria, indicating Fanconi syndrome. Both horses recovered with supportive therapy. This is the first report of acquired Fanconi syndrome in horses.

Syndrome de Fanconi transitoire chez des chevaux Quarter horse. Deux chevaux Quarter horse ayant subi une perte de poids étaient atteints de glucosurie, d'euglycémie et d'une acidose métabolique légère suggérant un défaut tubulaire rénal proximal. De nouveaux tests ont révélé une amino-acidurie, une acidurie lactique et une euglycémie généralisée transitoire, indicatif du syndrome de Fanconi. Les deux chevaux se sont rétablis avec une thérapie de soutien. Il s'agit du premier rapport du syndrome de Fanconi acquis chez les chevaux.(Traduit par Isabelle Vallières).

Intra-abdominal hyaluronan concentration in peritoneal fluid of horses with sudden signs of severe abdominal pain.

OBJECTIVE: To determine hyaluronan concentrations in peritoneal fluid from healthy horses and horses with sudden signs of severe abdominal pain and to identify the cellular sources of hyaluronan within the peritoneal cavity. ANIMALS: 7 client-owned horses that were evaluated for sudden signs of severe abdominal pain, 6 healthy teaching horses, and 13 euthanized horses (11 with no abdominal disease and 2 that had undergone abdominal surgery 2 weeks previously for a different study). PROCEDURES: Abdominal fluid was collected from the client-owned and teaching horses. Hyaluronan concentrations were determined with an ELISA. Equine mesothelial cells were aseptically harvested from euthanized horses immediately after euthanasia, cultured, and processed for western blot immunoassays to detect expression of the following mesothelial cell markers: cytokeratins 8 and 18, vimentin, calretinin, mesothelin, and CD44. A reverse transcriptase-PCR assay was used to detect genetic expression of hyaluronan synthase-2 (HAS-2) from cultured and native equine tissue. RESULTS: The mean ± SD abdominal hyaluronan concentration in peritoneal fluid from horses with signs of abdominal pain (1,203.3 ± 46.3 ng/mL) was significantly greater than that in healthy horses (228.4 ± 167.3 ng/mL). Harvested cells were maintained, and immunoblotting analyses confirmed expression of the mesothelial markers. Gene expression of HAS-2 from cultured mesothelial cells and fibroblasts was confirmed. CONCLUSIONS AND CLINICAL RELEVANCE: Peritoneal hyaluronan concentration was much higher in horses with severe abdominal pain than in healthy horses. Cultured equine mesothelial cells and fibroblasts can produce hyaluronan through HAS-2. Future investigation should focus on establishing the effect of exogenous hyaluronan administration on mesothelial cell function in horses with abdominal disease.

Web-based documentation of clinical skills to assess the competency of veterinary students.

Kansas State University implemented a Web-based program to assess students' competency to perform technical skills during clinical rotations throughout the fourth year of the veterinary curriculum. The classes of 2009 and 2010 recorded a minimum number of procedures (104 and 103, respectively) from a menu of more than 220 recommended procedures. Procedures were categorized by species (small animal, equine, food animal) and disciplines (imaging, anesthesia, diagnostic medicine/necropsy). Ophthalmology was added as a fourth discipline for the class of 2010. Students recorded procedures into the Web-based system, including information about the patient, procedure performed, supervisor, and a self-assessment of performance. Faculty, staff, and house officers evaluated the procedures electronically by confirming that they witnessed the procedure and providing qualitative and written feedback. The class of 2009 recorded 18,492 procedures (M=171/student) and the class of 2010 recorded 16,935 procedures (M=158/student). Two students from each class (2009 and 2010) did not complete the minimum required skills during clinical rotations and returned to perform procedures immediately before (n=3) or immediately after (n=1) graduation to receive their diploma. The Web-based system captured a large number of assessments of technical competency performed in the clinical setting. The system provided students with formative feedback throughout the clinical year, ensured equitable distribution of procedural opportunities across the student body, and required minimal additional resources.

Pharmacokinetics and pharmacodynamics of dexamethasone after oral administration in apparently healthy horses.

OBJECTIVE: To assess pharmacokinetic and pharmacodynamic properties of dexamethasone administered PO as a solution or powder, compared with properties of dexamethasone solution administered IV, in apparently healthy horses. ANIMALS: 6 adult horses. PROCEDURES: Serum cortisol concentration for each horse was determined before each treatment (baseline values). Dexamethasone (0.05 mg/kg) was administered PO (in solution or powdered form) or IV (solution) to horses from which feed had or had not been withheld (unfed and fed horses, respectively). Each horse received all 6 treatments in random order at 2-week intervals; PO and IV administrations of dexamethasone were accompanied by IV or PO sham treatments, respectively. Plasma dexamethasone and serum cortisol concentrations were assessed at predetermined intervals. RESULTS: Maximum plasma dexamethasone concentration after PO administration of powdered dexamethasone in unfed horses was significantly higher than the maximum plasma concentration after PO administration of dexamethasone solution in unfed or fed horses. Mean bioavailability of dexamethasone ranged from 28% to 66% but was not significantly different among horses receiving either formulation PO in the unfed or fed state. After dexamethasone treatment PO or IV, serum cortisol concentrations were significantly less than baseline at 1 to 72 hours in unfed horses and at 2 to 48 hours in fed horses. CONCLUSIONS AND CLINICAL RELEVANCE: PO or IV administration of dexamethasone resulted in suppression of cortisol secretion in unfed and fed adult horses; the magnitude of suppression did not differ among treatment groups, and serum cortisol concentrations returned to baseline after 48 to 72 hours.

The effect of differing Audience Response System question types on student attention in the veterinary medical classroom.

The purpose of this study was to evaluate the ability of specific types of multiple-choice questions delivered using an Audience Response System (ARS) to maintain student attention in a professional educational setting. Veterinary students (N=324) enrolled in the first three years of the professional curriculum were presented with four different ARS question types (knowledge base, discussion, polling, and psychological investment) and no ARS questions (control) during five lectures presented by 10 instructors in 10 core courses. Toward the end of the lecture, students were polled to determine the relative effectiveness of specific question types. Student participation was high (76.1%+/-2.0), and most students indicated that the system enhanced the lecture (64.4%). Knowledge base and discussion questions resulted in the highest student-reported attention to lecture content. Questions polling students about their experiences resulted in attention rates similar to those without use of ARS technology. Psychological investment questions, based on upcoming lecture content, detracted from student attention. Faculty preparation time for three ARS questions was shorter for knowledge base questions (22.3 min) compared with discussion and psychological investment questions (38.6 min and 34.7 min, respectively). Polling questions required less time to prepare (22.2 min) than discussion questions but were not different from other types. Faculty stated that the investment in preparation time was justified on the basis of the impact on classroom atmosphere. These findings indicate that audience response systems enhance attention and interest during lectures when used to pose questions that require application of an existing knowledge base and allow for peer interaction.

Determination of internal control for gene expression studies in equine tissues and cell culture using quantitative RT-PCR.

Quantitative reverse transcription polymerase chain reaction (RT-PCR) has become a basic, reliable and sensitive modern technique, in both biological research and clinical diagnosis, for investigation of gene expression and validation of cDNA microarray analysis. Accurate mRNA quantification using quantitative RT-PCR commonly requires data normalization through stable housekeeping genes (HKGs). Selection of HKGs for data normalization is critical for accurate mRNA quantification. Our objective was to evaluate a set of candidate HKGs as internal controls for gene expression studies using quantitative RT-PCR in equine tissues and cell culture. One-step quantitative RT-PCR for 6 HKGs was performed using total RNA from equine tissue samples and cultured peripheral blood mononuclear cells (PBMCs). The stability of HKGs was mainly evaluated by analysis of variance, analyses of the standard deviation and coefficient of variation of Ct, and change of Ct of HKGs between control and treated samples. 18S rRNA consistently showed the smallest standard deviation and coefficient of variation, and the least change of Ct between control and treated samples, thus was identified as the most stable HKG for mRNA data normalization in quantitative RT-PCR for studying gene expression in equine tissues and cultured PBMCs.

Investigation of antigen specific lymphocyte responses in healthy horses vaccinated with an inactivated West Nile virus vaccine.

West Nile virus (WNV) is a single-stranded, enveloped RNA virus capable of causing encephalitic disease in horses. Unvaccinated horses are at risk for developing WNV disease in endemic geographic regions. Effective vaccination reduces disease frequency and diminishes disease severity in vaccinated individuals that become infected with WNV. Recent data indicate CD4+ lymphocytes are required for effective protection against disease; in particular, cross talk between CD4+ and CD8+ lymphocytes must be functional. The objective of this project was to investigate immune responses in horses throughout a series of three vaccinations using a commercial inactivated vaccine under natural conditions. Immune responses to vaccination were determined by neutralizing antibody titers with plaque reduction neutralization test (PRNT), IgM titer (capture ELISA), WNV specific antibody Ig subclass responses, WNV lymphocyte proliferative responses and intracellular cytokine expression. Horses were vaccinated with a series of three vaccines at 3-week intervals using an inactivated product. An initial measure of immune activation following vaccination was determined by evaluating changes in lymphocyte cytokine expression. Interferon (IFN) gamma and interleukin (IL)-4 expressing CD4+ lymphocytes significantly increased 14 days following initial vaccination compared to unvaccinated horses (P<0.05). IFN-gamma expressing CD8+ lymphocytes also increased and remained elevated for 110 days. Antigen specific lymphocyte proliferative responses were significantly increased up to 90 days following the third vaccination (P<0.05). As expected, vaccinated horses produced increased neutralizing antibody based on PRNT data and WNV antigen-specific Ig subclass responses compared with unvaccinated horses (P<0.05). Our data indicate that WNV vaccination with an inactivated product effectively induced an antigen-specific antibody responses, as well as CD4+ and CD8+ lymphocyte activation.

Molecular cloning and characterization of equine Toll-like receptor 9.

Innate immunity relies on a series of germline-encoded pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), to detect conserved microbial components. TLR9 is typically expressed intracellularly in immune cells such as dendritic cells and recognizes unmethylated bacterial or viral cytosine-phosphate-guanine DNA (CpG-DNA). To investigate innate immune responses through TLR9 signaling pathway in horses, we cloned and characterized equine TLR9. Protein sequence analysis shows that equine TLR9 has a typically conserved cytosolic Toll/interleukin-1 receptor (TIR) domain, three leucine-rich repeat (LRR) motifs, with greater than 82% identity to human, monkey, bovine, canine, feline, porcine and ovine orthologs. Equine TLR9 mRNA expression was characterized for spleen, lymph node, and peripheral blood leukocyte samples. Flow cytometric analysis of equine TLR9 expression using a cross-reactive TLR9 mAb identified high constitutive expression of equine TLR9 in PMNs, CD4(+) and CD8(+) T-lymphocytes as well as other leukocytes; similar to human TLR9 expression. The conservation of equine TLR9 and high expression profile in leukocytes suggests that equine TLR9 is a frequent target for unmethylated CpG-DNA, an essential mechanism for the activation of innate immunity.

Molecular cloning and characterization of equine NK-lysin.

NK-lysin is an antimicrobial peptide of cytotoxic and NK lymphocytes that has powerful antibacterial properties as well as antitumoral activity. Here we report the full-length cDNA and deduced amino acid sequence for equine NK-lysin. Equine NK-lysin is 67% identical to porcine NK-lysin, 53% identical to bovine NK-lysin and 41% identical to granulysin in amino acid sequence. Complete conservation of cysteine residues between equine, bovine and porcine NK-lysin suggests similar disulfide bonding patterns among these peptides. Equine NK-lysin has the most positive surface charge when compared with other homologues. Similar to expression profiles in other species, equine NK-lysin is constitutively transcribed in various lymphocytes that include CD4+ and CD8+ staining cells. These findings suggest that equine NK-lysin, similar in cDNA sequence to the porcine, bovine and human homologues may play a role in antimicrobial defense.

Equine beta-defensin-1: full-length cDNA sequence and tissue expression.

beta-Defensins are cysteine-rich endogenously produced antimicrobial peptides that play an important role in innate immune defense. Although, previous investigations have identified beta-defensins in several mammalian species, no reports have identified equine beta-defensins. Using a strategy of database searching for expressed sequence tags (EST) we identified putative expression of equine beta-defensins in hepatic tissue. Based on this information, sequence specific primers were designed for the equine gene enabling the identification of the full-length cDNA sequence of equine beta-defensin-1. Comparative analyses showed that equine beta-defensin-1 has 46-52% amino-acid identity with other beta-defensins, sharing the greatest identity with porcine beta-defensin-1. Complete conservation of cysteine residues was maintained between the species evaluated, and RT-PCR analysis revealed diverse mRNA tissue expression for equine beta-defensin-1. These data extend the repertoire of equine antimicrobial peptides and expand our understanding of equine innate immunity.

Use of dapsone in the treatment of Pneumocystis carinii pneumonia in a foal.

A 6-month-old male Quarter Horse was evaluated for chronic respiratory tract disease. Diagnostic investigations revealed pulmonary inflammation; Pneumocystis carinii was detected within macrophages. Lymphocyte subpopulation phenotyping and immunoglobulin concentration analysis were performed and results suggested immune suppression. Trimethoprim-sulfamethoxazole administration was initiated; the colt was discharged but was reexamined 8 days later because of profuse diarrhea and endotoxemia. Bacterial culture of feces recovered Salmonella spp resistant to trimethoprim-sulfamethoxazole, and a diagnosis of antimicrobial-associated colitis was made. Bilateral fibrinous hypopyon developed and was treated with topical medication and intracameral injections of human recombinant tissue plasminogen activator. Dapsone (3 mg/kg [1.4 mg/lb], PO, q 24 h; dose extrapolated from human data) was administered for treatment of P carinii pneumonia (56-day treatment period). The colt recovered from the pneumonia and diarrhea. Dapsone may be a useful adjunct to traditional treatment for P carinii pneumonia in horses or as a sole medication for horses that cannot tolerate other treatments.

Dexamethasone for treatment of multisystemic eosinophilic epitheliotropic disease in a horse.

A 4-year-old Paint mare was examined because of respiratory tract infection, dermatitis, and weight loss of 2 months' duration. Initial examination revealed generalized pruritic dermatitis, ocular and nasal discharges, and stranguria. Laboratory abnormalities included leukopenia and hypoalbuminemia. Further examination of the respiratory tract revealed grade III of IV pharyngitis and pyogranulomatous pneumonia. Endoscopic examination of the bladder revealed a prolific mass at the junction of the bladder and urethra. Hypoproteinemia was suspected to be caused by protein-losing enteropathy. On histologic examination, skin, rectal, pharyngeal, and urethral biopsy specimens were characterized by infiltration of eosinophils and lymphocytes, and a diagnosis of multisystemic eosinophilic epitheliotropic disease was made. The horse improved following treatment with dexamethasone, trimethoprim-sulfamethoxazole, and an antihistamine and was discharged after 19 days of hospitalization. Treatment with dexamethasone was continued for 4 weeks after hospitalization but was then discontinued. Eight months after discharge, the horse was performing as a pleasure horse and did not require any medical treatment. Multisystemic eosinophilic epitheliotropic disease is typically associated with a poor prognosis in horses. The dermatitis, protein-losing enteropathy, and lower respiratory tract disease in this horse were consistent with previous reports; however, pharyngitis and urethritis have not, to our knowledge, been previously reported in horses with this disease.

Increases in cytokine and antimicrobial peptide gene expression in horses by immunomodulation with Propionibacterium acnes.

Immunomodulation with Propionibacterium acnes is used for prophylaxis of respiratory disease or in horses suffering from chronic pulmonary inflammation; however, the mechanism for this response is poorly understood. Semiquantitative reverse transcriptase-polymerase chain reaction assays were used to evaluate gene expression of interleukin (IL)-2, IL-10, interferon (IFN)-gamma, and NK-lysin in healthy horses treated with P. acnes. Findings in the study indicated that horses treated with a P. acnes-based immunomodulator exhibited increased IFN-gamma and NK-lysin gene expression in peripheral blood mononuclear cells. These results suggest that part of the immunostimulating properties of a P. acnes-based immunomodulator is derived from enhanced gene expression of the type-1 cytokine IFN-gamma and NK-lysin, an antimicrobial peptide.

Flow cytometry: clinical applications in equine medicine.

The use of flow cytometry in veterinary diagnostics is becoming a valuable clinical tool with a broad range of applications. Physical characteristics of cells can be determined by the flow cytometer laser and electronics through the measurement of changes in light scatter properties. Other components and functions of cells can be defined through the application of fluorochrome dyes that have an affinity for cellular components. Traditionally, common clinical applications are immunophenotyping of cells of the hematopoietic system with fluorescent-labeled antibodies raised against specific cell surface proteins. Other approaches have been used to elucidate changes in cell function and DNA content. This review is intended to provide the reader with the fundamental uses of flow cytometry. Examples of clinical applications in equine patients include immune-mediated hemolytic anemia, immune-mediated thrombocytopenia (IMT), chronic inflammatory disease, and neoplasia.

Cathelicidins: microbicidal activity, mechanisms of action, and roles in innate immunity.

Antimicrobial peptides are important host-defense molecules of innate immunity. Cathelicidins are a diverse family of potent, rapidly acting and broadly effective antimicrobial peptides, which are produced by a variety of cells. This review examines the classification, antimicrobial spectrum, mechanism of action, and regulation of cathelicidins.