Review of the clinical diagnosis of sacroiliac dysfunction in horses - Challenges and limitations.

Sacroiliac dysfunction (SID) is a condition seen in horses associated with poor performance that affects hind limb gait and impulsion. The condition comprises pain and dysfunction but there lacks clarity around the aetiopathogenesis and whether SID encompasses abnormal joint pathology, abnormal joint movement, abnormal regional biomechanical function, joint laxity and pain, or various combinations of these that may vary over time. Clinical assessment remains challenging for equine clinicians due to the deep location of the sacroiliac joint (SIJ) and surrounding structures which limits access for palpation, diagnostic imaging and joint-specific injection. There is no recognised single reference standard diagnostic test for SID. Clinical diagnosis has been based on ruling out other causes of hind limb lameness, along with combinations of ultrasonography, scintigraphy and periarticular anaesthesia of the SIJ. Recent studies have highlighted the lack of specificity of injections targeting the SIJ, with significant dispersal of injectate into surrounding structures including around the lumbosacral joint (LSJ). Advanced imaging modalities such as computed tomography offers promise for assessment of the structure and pathology of the SIJ and surrounding bony structures. However, there is a need to improve the understanding of the significance of anatomic variation of the sacroiliac region structures, with recent studies reporting detailed anatomic variation in groups of horses with and without SID. There are also limitations around functional assessment of the joint which is still largely reliant on a thorough clinical examination. This review aims to present an update on clinical approaches to the diagnosis of horses with SID, and to consider the challenges and limitations.

Use of physical examination, electrocardiography, radiography, and biomarkers to predict echocardiographic stage B2 myxomatous mitral valve disease in preclinical Cavalier King Charles Spaniels.

INTRODUCTION: Cavalier King Charles Spaniels (CKCS) are predisposed to developing myxomatous mitral valve disease (MMVD). Dogs with stage B2 MMVD benefit from medication. OBJECTIVES: To develop (1) breed-specific cut-offs for individual screening tests and (2) predictive models utilizing physical examination (PE), ECG, radiograph, and blood-based biomarker variables in combination for identification of echocardiographic stage B2 MMVD in preclinical CKCS. ANIMALS: Adult, preclinical CKCS not receiving cardiac medications (N = 226). MATERIALS AND METHODS: Prospective, cross-sectional study. Enrolled CKCS underwent PE, ECG, radiography, Doppler blood pressure measurement, echocardiography, and biomarker testing. Dogs were grouped by MMVD stage using echocardiography only. The discriminatory ability of individual tests to identify stage B2 was assessed, and prediction models were developed using variables derived from four 'tests' (PE, ECG, radiography, and biomarkers). RESULTS: N-terminal pro-B-type natriuretic peptide (NT-proBNP) and radiographic vertebral heart size (VHS) had the best discriminatory ability of individual diagnostic tests to differentiate stage A/B1 CKCS from stage B2, with an area under the curve (AUC) of 0.855 and 0.843, respectively. An NT-proBNP ≥1138 pmol/L or a VHS ≥11.5 had high specificity for predicting stage B2 (90.1% and 90.6%, respectively). Prediction models incorporating variables from multiple tests had better discriminatory ability than single tests. The four-test prediction model had an AUC of 0.971. Three and two-test models had AUCs ranging between 0.925-0.959 and 0.895-0.949, respectively. CONCLUSIONS: Both NT-proBNP and VHS have good utility for predicting echocardiographic stage B2 MMVD in CKCS as individual tests. Prediction models incorporating multiple test variables have superior discriminatory ability.

Galectin-3 as a novel biomarker in cats with hypertrophic cardiomyopathy.

INTRODUCTION/OBJECTIVES: Galectin-3 (Gal-3) is a circulating biomarker of fibrosis. In humans, increased Gal-3 is predictive of myocardial fibrosis and adverse cardiac events. The aim of this study was to evaluate the potential for Gal-3 as a cardiac biomarker in cats with hypertrophic cardiomyopathy (HCM). MATERIALS AND METHODS: Eighty cats were enrolled (25 healthy cats with normal hearts, 35 with HCM American College of Veterinary Internal Medicine (ACVIM) stage B, and 21 with HCM ACVIM stage C). Each cat received a full echocardiogram, health panel, and total thyroxin level. Galectin-3 levels were measured for each enrolled patient. Troponin I and N-terminal pro-brain natriuretic peptide (NT-proBNP) were obtained for the majority of cats. Additionally, 17 ACVIM stage B cats underwent cardiac-gated magnetic resonance (CMR) imaging to assess myocardial extracellular volume (ECV), a noninvasive measure of myocardial fibrosis. RESULTS: Galectin-3 levels are increased in cats with HCM ACVIM stage B and C compared to healthy cats; however, no significant differences were detected between ACVIM stage B and ACVIM stage C cats. In HCM-affected cats, Galectin-3 showed statistically significant correlations with left atrial dimensions, left atrial:aorta ratio, and CMR-derived ECV. Quantitative NT-proBNP showed excellent discrimination between all groups and troponin I was able to discriminate between ACVIM stage C and normal cats, but not between other groups. CONCLUSIONS: Circulating Gal-3 levels are increased in cats with HCM and is positively correlated with left atrial dimensions and ECV in affected cats. Further studies evaluating the relationship between Gal-3, myocardial fibrosis, and clinical outcomes are warranted.

Inflammatory Arthritis Post Covid-19 Infection.

Presentation A 63-year-old man developed polyarthritis two months post recovery from COVID-19 infection. Diagnosis We concluded that the diagnosis was rheumatoid arthritis based upon raised inflammatory markers, positive rheumatoid factor and anti-cyclic citrullinated peptide antibodies. Treatment His symptoms improved with naproxen, corticosteroids, and methotrexate. Discussion We describe a patient with late onset rheumatoid arthritis possibly triggered or unmasked by COVID-19.

Analysis of Posterior Zirconia Crowns with Vertical Margin Preparations.

BACKGROUND: The interest in vertical crown preparation is growing and several variations are presented in the dental literature but limited clinical outcome data exists for teeth prepared for a vertical finish line. AIM: To audit clinical outcomes for 73 teeth after vertical preparation for knife-edge zirconia crowns. METHOD: Seventy-three teeth had knife-edge zirconia crowns placed after vertical finish line preparation. The outcomes for these teeth were analysed from a retrospective audit of clinical records. RESULTS: The mean follow-up time was 21 months (SD: 10 months; range: 6-40). One tooth had to be extracted due to fracture at cervical level. One crown had to be re-made after margin fracture during try-in. No subsequent endodontic treatment was needed for any of the prepared teeth. No change in alveolar bone height was noted on radiographic follow-up. The mean bleeding score for the crowned teeth was higher than the mean bleeding score for a control tooth, but this was not statistically significant. CONCLUSION: This retrospective evaluation has shown favourable outcomes for 72 teeth after vertical preparation for knife-edge crowns. Longer follow-up is needed but the present results show that the technique is a viable procedure with potential advantages.

Comparison of three two-dimensional echocardiographic methods of assessing left ventricular size in Doberman Pinschers.

INTRODUCTION: Compare three methods of obtaining linear left ventricular dimensions within the same Doberman Pinscher (DP). ANIMALS: One hundred and thirty-nine client-owned DP. MATERIALS AND METHODS: Linear left ventricular dimensions were measured using two-dimensional short-axis (Sx-2D), motion-mode short-axis (Sx-MM), and motion-mode long-axis (Lx-MM) methods, then left ventricular volumes were obtained using monoplane Simpson's method of discs (SMOD). A Friedman test with Dunn's multiple comparisons was used to compare differences between methods. Bias and correlation were evaluated via Bland-Altman and Spearman's correlation. Sensitivity and specificity for diagnosing occult dilated cardiomyopathy (DCM) compared to SMOD were determined. Coefficients of variation (CVs) were calculated for intra- and inter-observer measurement variability. RESULTS: There were significant differences between all linear dimensions in diastole and systole. Short-axis 2D measurements had significant bias compared with Sx-MM (diastole +1.19 mm, systole +1.65 mm) and Lx-MM (diastole +4.36 mm, systole +3.87 mm) as did Sx-MM compared with Lx-MM (diastole +3.17 mm, systole +2.22 mm). All linear dimensions had a moderate positive correlation with SMOD. The sensitivity and specificity of linear measurements to detect DCM were: Sx-2D (sensitivity 72.0%, specificity 88.5%), Sx-MM (sensitivity 52.0%, specificity 92.0%), and Lx-MM (sensitivity 37.5%, specificity 99.1%). All methods had acceptably low CV for intra- and inter-observer measurement variability. CONCLUSIONS: Results of this study suggest that linear measurements are repeatable and correlate with reference standard; however, there is a significant bias between measurements, and they should not be used interchangeably.

Evaluation of the Feasibility, Reliability, and Repeatability of Welfare Indicators in Free-Roaming Horses: A Pilot Study.

Validated assessment protocols have been developed to quantify welfare states for intensively managed sport, pleasure, and working horses. There are few protocols for extensively managed or free-roaming populations. Here, we trialed welfare indicators to ascertain their feasibility, reliability, and repeatability using free-roaming Carneddau Mountain ponies as an example population. The project involved (1) the identification of animal and resource-based measures of welfare from both the literature and discussion with an expert group; (2) testing the feasibility and repeatability of a modified body condition score and mobility score on 34 free-roaming and conservation grazing Carneddau Mountain ponies; and (3) testing a prototype welfare assessment template comprising 12 animal-based and 6 resource-based welfare indicators, with a total of 20 questions, on 35 free-roaming Carneddau Mountain ponies to quantify inter-assessor reliability and repeatability. This pilot study revealed that many of the indicators were successfully repeatable and had good levels of inter-assessor reliability. Some of the indicators could not be verified for reliability due to low/absent occurrence. The results indicated that many animal and resource-based indicators commonly used in intensively managed equine settings could be measured in-range with minor modifications. This study is an initial step toward validating a much-needed tool for the welfare assessment of free-roaming and conservation grazing ponies.

Interspecific Competition for Colonization of Maize Plants Between Fusarium proliferatum and Fusarium verticillioides.

Fusarium proliferatum and F. verticillioides are mycotoxin-producing, seedborne pathogens of maize. They are often asymptomatic in seed, eluding symptom-based detection. Experiments were conducted in nonsterile soil to determine whether interspecific competition influenced establishment in maize plants of an introduced isolate of F. proliferatum or F. verticillioides. Hygromycin-resistant, green fluorescent protein (GFP) transformed (GFP-tagged) F. proliferatum (F. proliferatum-green) and hygromycin-resistant, monomeric red fluorescent protein (mRFP) transformed (mRFP-tagged) F. verticillioides (F. verticillioides-red) strains were developed to provide molecular markers to track fungal establishment. Heat-killed Fusarium-free maize seed, colonized with F. proliferatum-green or F. verticillioides-red by immersion in a spore suspension for 16 h, served as the source of inoculum. The ability of F. proliferatum-green and F. verticillioides-red to colonize viable maize plants already colonized by the other species was determined. Maize plants were retrieved from soil after 14 days and DNA was extracted from three consecutive root segments and three consecutive stem segments. A TaqMan multiplex real-time quantitative PCR protocol was developed to identify and quantify F. proliferatum-green and F. verticillioides-red from each plant segment from each treatment; the experiment was repeated three times. This experiment confirmed that F. proliferatum-green and F. verticillioides-red effectively colonized roots and stems of the maize plant already colonized with the other species. Prior colonization of maize tissues by F. verticillioides-red (P = 0.6749) and other seedborne microorganisms (P = 0.1910) reduced but did not prevent subsequent colonization by F. proliferatum-green. Similarly, prior colonization of maize tissues by F. proliferatum-green (P = 0.7032) and other seedborne microorganisms (P = 0.1447) reduced but did not prevent subsequent colonization by F. verticillioides-red.

Reference intervals and echocardiographic findings in Leonberger dogs.

INTRODUCTION: Leonbergers are large-breed dogs believed to be predisposed to dilated cardiomyopathy, yet the normal echocardiographic measurements of these dogs are unknown. The aim of this study was to describe echocardiographic findings and propose breed-specific reference intervals (RIs) for Leonbergers. ANIMALS: A total of 42 clinically healthy dogs were involved in this study. MATERIALS AND METHODS: This study was designed as a cross-sectional study of 42 Leonberger dogs undergoing complete transthoracic echocardiographic studies at a national show. Reference intervals were determined using robust method with bootstrapping. The effects of age, sex, and body weight (BW) on linear and volumetric dimensions were evaluated by regression analysis. Intra- and interobserver variability were evaluated by coefficient of variation from 10 of the studies. RESULTS: There were significant linear correlations with BW for many of the cardiac variables evaluated. However, despite the statistical significance, most regression equations had small slopes, necessitating large changes in BW for small changes in two-dimensional echocardiographic variables. As such, this study resulted in breed-specific RIs irrespective of BW. Trace mitral insufficiency (n = 18), trace aortic insufficiency (n = 15), or both (n = 1) was identified in 81% of dogs in this population. Coefficients of variation were <9% for all two-dimensional, volumetric, and motion mode measurements. CONCLUSIONS: This study describes echocardiographic findings and provides RIs, which may be useful in echocardiographic evaluations of Leonbergers and may aid in early identification of heart disease within the breed.

Low-cost assembly of a cacao crop genome is able to resolve complex heterozygous bubbles.

Cacao (Theobroma cacao) is a tropical tree that produces the essential raw material for chocolate. Because yields have been stagnant, land use has expanded to provide for increasing chocolate demand. Assembled genomes of key parents could modernize breeding programs in the remote and under-resourced locations where cacao is grown. The MinION, a long read sequencer that runs off of a laptop computer, has the potential to facilitate the assembly of the complex genomes of high-yielding F1 hybrids. Here, we validate the MinION's application to heterozygous crops by creating a de novo genome assembly of a key parent in breeding programs, the clone Pound 7. Our MinION-only assembly was 20% larger than the latest released cacao genome, with 10-fold greater contiguity, and the resolution of complex heterozygosity and repetitive elements. Polishing with Illumina short reads brought the predicted completeness of our assembly to similar levels to the previously released cacao genome assemblies. In contrast to previous cacao genome projects, our assembly required only a small scientific team and limited reagents. Our sequencing and assembly methods could easily be adopted by under-resourced breeding programs, speeding crop improvement in the developing world.

Non-Dietary Correlates and Determinants of Plasma Lutein and Zeaxanthin Concentrations in the Irish Population.

OBJECTIVE: To investigate non-dietary correlates and determinants of plasma lutein (L) and zeaxanthin (Z) concentrations in The Irish Longitudinal Study on Ageing (TILDA) sample. DESIGN: Cross-sectional study. SETTING: Community dwelling adults in the Republic of Ireland (ROI). PARTICIPANTS: 3,681 participants aged 50 years and older. MEASUREMENTS: TILDA is a nationally representative prospective cohort study of community dwelling adults aged 50 years and over in the ROI. Demographic and health variables were collected during a face-to-face interview carried out in the home (n=8175), and a substantial proportion of these (n=5035; 62%) also attended a study visit in a health assessment centre. Blood samples collected at baseline (wave 1, the subject of the current study), were analysed for plasma concentrations of L and Z by reversed-phase high performance liquid chromatography, and macular pigment (MP) optical density was also measured (using customized heterochromatic flicker photometry). RESULTS: After excluding participants with eye disease, data from 3,681 participants were available for analysis. For this group of participants, plasma L and Z were inversely and significantly associated with body mass index (BMI), and were positively and significantly associated with MP, total cholesterol, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) (p<0.001, for all). Plasma L and Z were significantly lower in males, current smokers, participants reporting less physical exercise, and participants reporting lower levels of education (p<0.05, for all). Plasma L was significantly higher in participants reporting a family history of age-related macular degeneration (AMD) (p=0.001), and in the group of ≥75 years old (p<0.05). For each of these variables, the significant associations remained after controlling for other potential confounding variables. CONCLUSION: The findings of this large study indicate that plasma concentrations of L and Z were lower in association with indicators of a poor lifestyle (high BMI, tobacco use, and less physical exercise) and in association with lower education, indicating that modifying lifestyle in a positive way is likely to be reflected in higher concentrations of plasma carotenoids, with consequential and putative health benefits.

Repair of large osteochondritis dissecans lesions using a novel multilayered tissue engineered construct in an equine athlete.

Osteochondral lesions resulting from osteochondritis dissecans are problematic to treat and present a significant challenge for clinicians. The aims of this study were to investigate the use of a scaffold-assisted microfracture approach, employing a novel, multilayered, collagen-based, osteochondral graft substitute in the treatment of severe osteochondritis dissecans of both lateral femoral trochlear ridges in an equine athlete, and to assess the potential of this novel scaffold to enhance repair of the osteochondral unit. A 15 month-old female filly presented with large osteochondritis dissecans lesions involving both femoral lateral trochlear ridges. After routine arthroscopic debridement and microfracture of the subchondral bone, multilayered osteochondral defect repair scaffolds were implanted into the fragmentation beds in both left and right femoropatellar joints via mini-arthrotomies. Exploratory arthroscopy 5 months postimplantation revealed smooth cartilaginous repair tissue, contiguous with the adjacent cartilage, covering the defect. At 22-month follow up, the filly had no signs of lameness and was exercising at her intended level. Radiographically, although still slightly flattened, the femoral trochlear ridges were smooth, with no evidence of osteoarthritis. Ultrasonographically, the defects were filled with bone and covered with an overlying cartilaginous layer, with the trochlear ridge contour almost entirely restored. This report demonstrates the effective clinical use of this novel, multilayered, osteochondral defect repair scaffold in the treatment of osteochondritis dissecans of an equine athlete. The successful repair achieved here using this novel scaffold in an equine patient with large bilateral lesions shows the potential for clinical translation in the treatment of human patients presenting with osteochondral defects. Copyright © 2016 John Wiley & Sons, Ltd.

Development of simple sequence repeat (SSR) markers for discrimination among isolates of Fusarium proliferatum.

The plant pathogen Fusarium proliferatum has a wide host range and occurs worldwide. Many isolates of the fungus produce mycotoxins in plant tissues, which, if ingested, can cause harm to animals and humans. In 2008, an outbreak of salmon blotch of onions, caused by F. proliferatum, was detected in southern Israel. The source and distribution of the fungus in Israel were unknown. Inter-simple sequence repeats (ISSR) were used to identify repetitive motifs present in seven isolates of F. proliferatum from Israel, Germany and Austria. ISSR repeat motifs were, used to develop 17 simple sequence repeat (SSR) loci. Six of these SSR markers were polymorphic in and consistently amplified from ten isolates collected in Israel, Germany, Austria and North America, from cucumber, onion, garlic, maize, and asparagus. These six polymorphic SSR alleles included 5 to 12 copies of di-, tri, and pentanucleotide motifs and yielded six to 9 alleles each. Sixteen of the SSR loci were amplified at least one of the seven Fusarium species, F. verticillioides, F. thapsinum, F. subglutinans, F. andiyazi, F. globosum, F. fujikoroi and F. oxysporum. The data demonstrate that these SSRs can be used for characterization of F. proliferatum isolates from diverse hosts and geographic locations and that they are transferable to other species of Fusarium.

The Evolution of a Process for Selecting and Prioritizing Plant Diseases for Recovery Plans.

One element of the cost of dealing with invasive species in the United States is the recovery from the arrival of exotic plant pathogens. We review the development of a process used to prioritize plant diseases for the federally mandated United State Department of Agriculture National Plant Disease Recovery System. A team of university, government, and industry scientists worked together over a 10-year period to develop a science-based objective approach to the challenge of effectively preparing for recovery plans from introduced pathogens, when the timing of the introduction of any single disease is unknown. Over time, the process transitioned from ad hoc, in which recovery plans were written when the relevant experts were able to do so, to a formally organized group-prioritization effort from which emerged the concept of generic recovery plan templates for groups of pathogens and diseases that have similar biological characteristics, and therefore, similar management approaches. Key characteristics for each template were determined through a multivariate analysis for 14 plant diseases for which a recovery plan already existed. The process was validated by a larger group of 15 plant pathologists, for which results were compared with those scored by 14 subject matter experts.

Sustained supplementation and monitored response with differing carotenoid formulations in early age-related macular degeneration.

PURPOSE: To compare the impact of sustained supplementation using different macular carotenoid formulations on macular pigment (MP) and visual function in early age-related macular degeneration (AMD). PATIENTS AND METHODS: Sixty-seven subjects with early AMD were randomly assigned to: Group 1 (20 mg per day lutein (L), 0.86 mg per day zeaxanthin (Z); Ultra Lutein), Group 2 (10 mg per day meso-zeaxanthin (MZ), 10 mg per day L, 2 mg per day Z; Macushield; Macuhealth), Group 3 (17 mg per day MZ, 3 mg per day L, 2 mg per day Z). MP was measured using customised heterochromatic flicker photometry and visual function was assessed by measuring contrast sensitivity (CS) and best-corrected visual acuity (BCVA). AMD was graded using the Wisconsin Age-Related Maculopathy Grading System (AREDS 11-step severity scale). RESULTS: At 3 years, a significant increase in MP from baseline was observed in all groups at each eccentricity (P<0.05), except at 1.75° in Group 1 (P=0.160). Between 24 and 36 months, significant increases in MP at each eccentricity were seen in Group 3 (P<0.05 for all), and at 0.50° in Group 2 (P<0.05), whereas no significant increases were seen in Group 1 (P>0.05 for all). At 36 months, compared with baseline, the following significant improvements (P<0.05) in CS were observed: Group 2-1.2, 6, and 9.6 cycles per degree (c.p.d.); Group 1-15.15 c.p.d.; and Group 3-6, 9.6, and 15.15 c.p.d. No significant changes in BCVA, or progression to advanced AMD, were observed. CONCLUSION: In early AMD, MP can be augmented with a variety of supplements, although the inclusion of MZ may confer benefits in terms of panprofile augmentation and in terms of CS enhancement.

Using molecular tools to identify the geographical origin of a case of human brucellosis.

Although Malta is historically linked with the zoonosis brucellosis, there had not been a case of the disease in either the human or livestock population for several years. However, in July 2013 a case of human brucellosis was identified on the island. To determine whether this recent case originated in Malta, four isolates from this case were subjected to molecular analysis. Molecular profiles generated using multilocus sequence analysis and multilocus variable number tandem repeat for the recent human case isolates and 11 Brucella melitensis strains of known Maltese origin were compared with others held on in-house and global databases. While the 11 isolates of Maltese origin formed a distinct cluster, the recent human isolation was not associated with these strains but instead clustered with isolates originating from the Horn of Africa. These data was congruent with epidemiological trace-back showed that the individual had travelled to Malta from Eritrea. This work highlights the potential of using molecular typing data to aid in epidemiological trace-back of Brucella isolations and assist in monitoring of the effectiveness of brucellosis control schemes.

Evolution of equine influenza virus in vaccinated horses.

Influenza A viruses are characterized by their ability to evade host immunity, even in vaccinated individuals. To determine how prior immunity shapes viral diversity in vivo, we studied the intra- and interhost evolution of equine influenza virus in vaccinated horses. Although the level and structure of genetic diversity were similar to those in naïve horses, intrahost bottlenecks may be more stringent in vaccinated animals, and mutations shared among horses often fall close to putative antigenic sites.