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Background: The association of microbiota with clinical outcomes and the taxa associated with colitis in horses remains generally unknown. Objectives: Describe the fecal microbiota of horses with colitis and investigate the association of the fecal microbiota with the development of laminitis and survival. Animals: Thirty‐six healthy and 55 colitis horses subdivided into laminitis (n = 15) and non‐laminitis (n = 39, 1 horse with chronic laminitis was removed from this comparison) and survivors (n = 27) and nonsurvivors (n = 28). Methods: Unmatched case‐control study. The Illumina MiSeq platform targeting the V4 region of the 16S ribosomal RNA gene was used to assess the microbiota. Results: The community membership (Jaccard index) and structure (Yue and Clayton index) were different (analysis of molecular variance [AMOVA]; P <.001) between healthy and colitis horses. The linear discriminant analysis effect size (LEfSe; linear discriminant analysis [LDA] >3; P <.05) and random forest analyses found Enterobacteriaceae, Lactobacillus, Streptococcus, and Enterococcus enriched in colitis horses, whereas Treponema, Faecalibacterium, Ruminococcaceae, and Lachnospiraceae were enriched in healthy horses. The community membership and structure of colitis horses with or without laminitis was (AMOVA; P >.05). Enterobacteriaceae, Streptococcus, and Lactobacillus were enriched in horses with laminitis (LDA > 3; P <.05). The community membership (AMOVA; P =.008) of surviving and nonsurviving horses was different. Nonsurviving horses had an enrichment of Enterobacteriaceae, Pseudomonas, Streptococcus, and Enterococcus (LDA >3; P <.05). Conclusion and Clinical Importance: Differences in the microbiota of horses with colitis that survive or do not survive are minor and, similarly, the microbiota differences in horses with colitis that do or do not develop laminitis are minor. [ABSTRACT FROM AUTHOR]

This study evaluated the impact of an algorithm targeting antimicrobial therapy of diarrhoeic calves on the incidence of diarrhoea, antimicrobial treatment rates, overall mortality, mortality of diarrhoeic calves and changes in the faecal microbiota. The algorithm was designed to target antimicrobial therapy in systemically ill calves from on two dairy farms. Retrospective (farm 1: 529 calves; farm 2: 639 calves) and prospective (farm 1: 639 calves; farm 2: 842 calves) cohorts were examined for 12 months before and after implementation of the algorithm. The Mantel–Haenszel test and Kaplan–Meier survival curves were used to assess the cumulative incidence risk (CIR) and time to development of each outcome before and after implementation of the algorithm. The CIR of antimicrobial treatment rates was 80% lower after implementation of the algorithm on both farms (CIR 0.19, 95% confidence interval 0.17–0.21). There was no difference in the CIR of overall mortality, but the CRI for mortality of diarrhoeic calves was lower in the period after implementation of the algorithm on one farm. The faecal microbiota of 15 healthy calves from both farms at each time period were characterised using a sequencing platform targeting the V4 region of the 16S rRNA gene. On both farms, there were significant differences in community membership and structure (parsimony P < 0.001). Use of the algorithm for treatment of diarrhoeic calves reduced antimicrobial treatment rates without a negative impact on the health of calves. However, the experimental design did not take into account the potential confounding effects of dietary changes between the study periods. [ABSTRACT FROM AUTHOR]

Theiler's disease (TD) is a (sub-)acute hepatitis in adult horses and one of the most common causes of acute hepatic failure. Recent findings indicate that equine parvovirus hepatitis (EqPV-H) likely causes TD and that its transmission occurs via iatrogenic and/or natural routes. Following the death of an EqPV-H positive mare with TD, close-contact mares and foals in the same paddock were monitored to evaluate if there was any evidence of EqPV-H. For this purpose, the serum of close contact horses was examined 6 and 42 days after the mare's death for the presence of EqPV-H DNA and changes in liver-associated serum biochemical parameters. The foals had higher EqPV-H viral loads than the mares. Apart from the mare that was euthanized, none of the horses included in this study showed signs of severe disease and nor did they have particularly elevated liver enzymes. Nucleotide sequence analysis revealed no major differences between the viral DNA detected in the serum of the dead mare and any of the in-contact horses. In conclusion, our data confirmed previous findings that horizontal transmission of EqPV-H may occur through close contact between horses. • Horizontal transmission of EqPV-H to close contact horses. • Foals display higher viral loads compared to mares. • None of the horses showed signs of severe disease with elevated liver enzymes. [ABSTRACT FROM AUTHOR]

Clostridioides difficile (formerly Clostridium difficile) is associated with colitis in foals and mature horses. C. difficile exposes specific phosphorylated polysaccharides (PSs), named PS-I, PS-II and PS-III. These cell-surface PSs are potential vaccine targets, especially the hexasaccharide phosphate PS-II, that has been found in all C. difficile ribotypes examined. Since we previously identified anti-PS-II circulating antibodies in horses, we postulated that vaccinating foals with PS-II may prevent colonization by C. difficile. In this study, we aim to evaluate the IgM antibody responses in foals to PS-II. To evaluate the reactogenicity and immunogenicity of C. difficile PS-II in foals, three-to four-month-old foals were vaccinated intramuscularly three times at intervals of three weeks with 100 μg/dose (3 foals) or 500 μg/dose (3 foals) of purified PS-II antigen with aluminum hydroxide adjuvant, or with a placebo preparation (2 foals) containing adjuvant alone. No injection site swelling, pain or fever was observed after vaccination. Two of the three foals receiving 100 μg/dose, and three out of three foals receiving 500 μg/dose of PS-II responded with increases in serum IgM antibodies. No control foals that received the placebo had IgM responses to PS-II. There was a trend towards a higher response rate in foals receiving 500 μg PS-II one week after second vaccination when compared to control foals and towards higher concentrations of serum IgM antibodies in foals receiving 500 μg PS-II. No adverse reactions were observed following vaccination with PS-II in foals; Serum IgM immune responses were induced by vaccination. A polysaccharide-based vaccine for C. difficile in horses deserves further investigation. • Clostridioides difficile polysaccharide PS-II is immunogenic in foals. • Vaccination of foals with C. difficile PS-II raises anti-PS-II serum IgM antibodies. • No adverse reactions were observed following vaccination with PS-II in foals. [ABSTRACT FROM AUTHOR]

The intestinal tract houses one of the richest and most complex microbial populations on the planet, and plays a critical role in health and a wide range of diseases. Limited studies using new sequencing technologies in horses are available. The objective of this study was to characterize the fecal microbiome of healthy horses and to compare the fecal microbiome of healthy horses to that of horses with undifferentiated colitis. A total of 195,748 sequences obtained from 6 healthy horses and 10 horses affected by undifferentiated colitis were analyzed. Firmicutes predominated (68%) among healthy horses followed by Bacteroidetes (14%) and Proteobacteria (10%). In contrast, Bacteroidetes (40%) was the most abundant phylum among horses with colitis, followed by Firmicutes (30%) and Proteobacteria (18%). Healthy horses had a significantly higher relative abundance of Actinobacteria and Spirochaetes while horses with colitis had significantly more Fusobacteria. Members of the Clostridia class were more abundant in healthy horses. Members of the Lachnospiraceae family were the most frequently shared among healthy individuals. The species richness reported here indicates the complexity of the equine intestinal microbiome. The predominance of Clostridia demonstrates the importance of this group of bacteria in healthy horses. The marked differences in the microbiome between healthy horses and horses with colitis indicate that colitis may be a disease of gut dysbiosis, rather than one that occurs simply through overgrowth of an individual pathogen. [ABSTRACT FROM AUTHOR]

Abstract: Clostridium difficile is an important cause of diarrhea in horses, causing sporadic and epidemic disease of varying severity. This study evaluated the molecular characteristics of 48 C. difficile isolates recovered from diarrheic horses admitted to a veterinary hospital by using PCR-ribotyping and toxin gene profile. Additionally, feces were tested for the presence of C. difficile toxin A/B via enzyme immunosorbant assay (EIA) in 38 horses. The toxin genes tcdA, tcdB and cdtB were present in 27 (56.25%), 35 (72.91%) and 2 (4.1%) strains, respectively. Eight isolates (16.6%) were A−B+ variants. Thirteen of forty-eight isolates (27.0%) did not posses any toxin genes (A−B−CDT−). A positive EIA result was reported in 17 (44%) of the cases. There was no association between the presence of different ribotypes or strains and toxin gene(s) profiles and the clinical outcome. [Copyright &y& Elsevier]

Case Description—4 racehorses were examined because of markedly abnormal behavior following administration of fluphenazine decanoate. Clinical Findings—Clinical signs included restlessness, agitation, profuse sweating, hypermetria, aimless circling, intense pawing and striking with the thoracic limbs, and rhythmic swinging of the head and neck alternating with episodes of severe stupor. Fluphenazine was detected in serum or plasma from all 4 horses. The dose of fluphenazine decanoate administered to 3 of the 4 horses was within the range (25 to 50 mg) routinely administered to adult humans. Treatment and Outcome—In 2 horses, there was no response to IV administration of diphenhydramine hydrochloride, but the abnormal behavior in these 2 horses appeared to resolve following administration of benztropine mesylate, and both horses returned to racing. The other 2 horses responded to diphenhydramine administration. One returned to racing. The other was euthanized because of severe neurologic signs, respiratory failure, and acute renal failure. Clinical Relevance—Findings indicate that adverse extrapyramidal effects may occur in horses given fluphenazine decanoate. These effects appear to be unpredictable and may be severe and life threatening. Use of fluphenazine decanoate as an anxiolytic in performance horses is not permitted in many racing and horse show jurisdictions, and analytic procedures are now available to detect the presence of fluphenazine in serum or plasma. [ABSTRACT FROM AUTHOR]

Simple Summary: DNA sequencing has been used to characterize the intestinal microbiota of horses. However, all published studies are based on Illumina technology to date because it is widely accessible, but a major limitation is the short bacterial DNA fragment sequenced. Other long-read sequencing technologies (i.e., PacBio) can classify bacteria at the species level and therefore are more meaningful for the identification of specific changes in the gut after the use of antimicrobials, for example. The objectives of this study were to characterize the gut microbiota of horses at the species level before and after trimethoprim sulfadiazine administration and to compare results with short-read Illumina sequencing. PacBio sequencing failed to classify the equine intestinal microbiota at the species level but detected greater richness and less unclassified bacteria compared to Illumina sequencing. Bacteroidetes were the most abundant bacteria in the feces, followed by Firmicutes and Fibrobacteres in this study. Further studies should focus on improving databanks for equine studies. All current studies have used Illumina short-read sequencing to characterize the equine intestinal microbiota. Long-read sequencing can classify bacteria at the species level. The objectives of this study were to characterize the gut microbiota of horses at the species level before and after trimethoprim sulfadiazine (TMS) administration and to compare results with Illumina sequencing. Nine horses received TMS (30 mg/kg) orally for 5 days twice a day to induce dysbiosis. Illumina sequencing of the V4 region or full-length PacBio sequencing of the 16S rRNA gene was performed in fecal samples collected before and after antibiotic administration. The relative abundance and alpha diversity were compared between the two technologies. PacBio failed to classify the equine intestinal microbiota at the species level but confirmed Bacteroidetes as the most abundant bacteria in the feces of the studied horses, followed by Firmicutes and Fibrobacteres. An unknown species of the Bacteroidales order was highly abundant (13%) and deserves further investigation. In conclusion, PacBio was not suitable to classify the equine microbiota species but detected greater richness and less unclassified bacteria. Further efforts in improving current databanks to be used in equine studies are necessary. [ABSTRACT FROM AUTHOR]

Background: This study aimed to describe and compare therapeutic approaches for horses with acute diarrhea in different geographic regions worldwide. Methods: Clinical information was retrospectively collected from diarrheic horses presented to participating institutions between 2016 and 2020, including fluid therapy on admission, antimicrobial drugs, probiotics, anti-endotoxic medications, anti-inflammatory drugs, gastroprotectants, digital cryotherapy, and toxin-binding agents. Seasonal and geographic differences were investigated. Results: 1438 horses from 26 participating hospitals from 5 continents were included. On admission, 65% (926/1419) of horses were administered a fluid bolus. Antimicrobial drugs were administered to 55% (792/1419) within the first 24 hours of admission, with penicillin and gentamicin being the most used combination (25%, 198/792). Horses with leukopenia (OR: 2.264, 95%CI: 1.754 to 2.921; P<0.001) or meeting systemic inflammatory response syndrome criteria (OR: 2.542, 95%CI: 1.919 to 3.368; P<0.001) had higher odds of being administered antimicrobial drugs. Other treatments administered included probiotics (15%, 215/1438), polymyxin B (13%; 187/1438), pentoxifylline (8%; 118/1438), gastroprotectants (44%; 626/1419), digital cryotherapy (34%; 489/1435), plasma transfusion (13%; 182/1410) and toxin-binding agents (36%; 515/1438). Limitations: Due to the retrospective nature of the study, the rationale for treatment decisions was unavailable, and associations with survival could not be evaluated. Conclusions: Treatments varied between hospitals from different geographic regions. Prospective clinical trials are required to evaluate the effects of various treatments on survival. [ABSTRACT FROM AUTHOR]

Simple Summary: Acute diarrhea (colitis) is a major problem in adult horses and the role of the intestinal bacteria (microbiota) is still poorly understood in this species. The aim of this study was to compare the mucosal and luminal content microbiota of the cecum and colon of healthy and diarrheic horses. We concluded that microbial dysbiosis (changes in the normal microbiota composition) occurs in horses with colitis at different levels of the intestinal tract and microbiota composition is different between the mucosa and luminal content of diarrheic horses. Changes in species associated with dysbiosis could be used in the future for disease diagnosis, prognosis and treatment of equine colitis. The aim of this study was to compare the mucosal and luminal content microbiota of the cecum and colon of healthy and diarrheic horses. Marked differences in the richness and in the community composition between the mucosal and luminal microbiota of the cecum and large colon of horses with colitis were observed. Microbial dysbiosis occurs in horses with colitis at different levels of the intestinal tract, and microbiota composition is different between the mucosa and luminal content of diarrheic horses. The changes in some key taxa associated with dysbiosis in the equine intestinal microbiota, such as Escherichia, Fusobacterium and Lactobacillus, deserve further inquiry in order to determine their utility for disease diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Background: Increased concentration of haptoglobin (Hp) in serum is associated with survival of critically ill humans and horses. High serum activity of liver‐derived enzyme is associated with sepsis in children and foals. Hypothesis/Objectives: Investigate whether admission serum Hp and glutamic dehydrogenase (GLDH) are associated with systemic inflammatory response syndrome (SIRS) and survival of sick calves. Animals: One hundred two calves. Methods: Retrospective cross‐sectional study. Electronic medical records from all calves <30 days of age admitted to a teaching hospital for 8 years were reviewed. The signalment, clinicopathological findings, the presence of SIRS, final diagnosis, hospitalization time and outcome were recorded. A Cox proportional hazard ratio (HzR) were calculated to assess the association between clinicopathological variables and survival to discharge. Results: Serum Hp concentrations were similar between SIRS (0.29 g/L; range, 0.05‐3.6) and non‐SIRS calves (0.22 g/L; range, 0‐4.2; P =.62). GLDH activity was similar between SIRS (12 U/L; range, 1‐1025) and non‐SIRS calves (9 U/L; range, 2‐137; P =.2). Absent suckle reflex (HzR: 6.44, 95% CI: 1.44‐28.86), heart rate (HR) < 100 beats per minute (bpm; HzR: 12.2; 95% CI: 2.54‐58.62), HR > 140 bpm (HzR: 3.59, 95% CI: 1.05‐12.33), neutrophil count <1.7 × 109/L (HzR: 7.36; 95% CI: 2.03‐26.66) and increased gamma‐glutamyl transferase activity (every 50‐unit, HzR: 1.12; 95% CI: 1.03‐1.21) were predictive of nonsurvival. Conclusions and Clinical Importance: The use of Hp and GLDH for prediction of survival in sick calves cannot be recommended at this time. [ABSTRACT FROM AUTHOR]

The article presents a letter to the editor about Clostridium difficile-associated disease.

A letter to the editor is presented in response to the article "Effects of Alkalinisation and Rehydration on Plasma Potassium Concentration in Nenonatal Calves with Diarrhea," by F. M. Trefz et al. in the March 2015 issue.

OBJECTIVE: To describe histologic findings in the small airways during postmortem examination of actively racing horses and to quantify the degree of airway inflammation by use of a semiquantitative scoring system. SAMPLE: Lung tissues obtained from 95 horses (Thoroughbreds, Standardbreds, and Quarter Horses) that had been actively racing or training. PROCEDURES: 10 standardized lung sections were obtained during postmortem examination of 37 horses that had been actively racing or training, and 2 histologic sections of the caudodorsal lung lobes previously collected from each of 58 horses that died or were euthanized during racing or training were also obtained. Sections were evaluated by use of a validated histologic small airway scoring system. RESULTS: Scores for inflammatory cells, smooth muscle, and hemosiderin typically were high. Signalment and cause of death were not significant predictors of lung scores. Lung sample location was a significant predictor, with the highest scores in the caudal and dorsal sections. Inflammatory cell infiltration in peribronchiolar tissues, smooth muscle hyperplasia, and hemosiderin (prevalence of 86%, 98%, and 80%, respectively) were common findings in lungs of these horses, with the caudodorsal regions more severely affected. Correlation was moderate between smooth muscle hyperplasia and inflammatory cell infltration, with minimal correlation between hemosiderin and inflammatory cell infiltration. CONCLUSIONS AND CLINICAL RELEVANCE: Inflammatory airway disease has been identified by use of bronchoalveolar lavage in young athletic horses throughout the world. In the study reported here, pathological changes were detected in the wall of small airways of horses that were actively training or racing. [ABSTRACT FROM AUTHOR]

A case study involves a 2-day-old Appaloosa filly presenting with straining and abdominal distension. Diagnostic imaging via abdominal ultrasonography reveals a large amount of homogeneous anechoic free peritoneal fluid with aggregation of the intestinal tube, indicative of uroperitoneum. Differential diagnoses include uroperitoneum, Foley, neonatal foal, medical management and equine.

Next generation sequencing has demonstrated that alpha diversity of the fecal microbiota is significantly altered in horses with typhlocolitis. The objective of this study was to evaluate the bacterial composition of the cecum content of horses with and without typhlocolitis through direct and culture-enriched 16S gene sequencing of six healthy horses and six horses with acute typhlocolitis; a case-control study design. Cecal content was collected after euthanasia. An aliquot was used for direct 16S gene sequencing. Another was serially diluted with brain heart infusion (BHI) and plated onto five different agar media. All culture medias, except for MacConkey, were incubated anaerobically. Bacterial colonies were harvested in bulk and used for DNA extraction, 16S PCR amplification, and sequenced using the Illumina MiSeq platform. Predominant phyla in healthy and diseased horses were Firmicutes, followed by Bacteroidetes in all cultured medias, except for MacConkey agar, in which Proteobacteria was the dominant phylum. Greater bacterial richness was identified in sequenced cecal contents as compared to cultured plates (P < 0.05). Culture-enriched molecular profiling combined with 16S rRNA gene sequencing offer an alternative method for the study of the gut microbiota of horses. For direct cecum content 16S gene amplification, the alpha diversity indices were lower in diarrheic horses compared to healthy horses (P < 0.05). A higher relative abundance of Fusobacteriota was found in 2/6 samples from diarrheic horses. The role of Fusobacteriota in equine colitis deserves investigation. [ABSTRACT FROM AUTHOR]

Alveolar macrophages (AMs) are the predominant innate immune cell in the distal respiratory tract. During inflammatory responses, AMs may be supplemented by blood monocytes, which differentiate into monocyte-derived macrophages (MDMs). Macrophages play important roles in a variety of common equine lower airway diseases, including severe equine asthma (SEA). In an experimental model, an inhaled mixture of Aspergillus fumigatus spores, lipopolysaccharide, and silica microspheres (FLS), induced SEA exacerbation in susceptible horses. However, whether equine AMs and MDMs have differing immunophenotypes and cytokine responses to FLS stimulation is unknown. To address these questions, alveolar macrophages/monocytes (AMMs) were isolated from bronchoalveolar lavage fluid and MDMs derived from blood of six healthy horses. Separately, AMMs and MDMs were cultured with and without FLS for six hours after which cell surface marker expression and cytokine production were analyzed by flow cytometry and a bead-based multiplex assay, respectively. Results showed that regardless of exposure conditions, AMMs had significantly higher surface expression of CD163 and CD206 than MDMs. Incubation with FLS induced secretion of IL-1β, IL-8, TNF-α and IFN-γ in AMMs, and IL-8, IL-10 and TNF-α in MDMs. These results suggest that AMMs have a greater proinflammatory response to in vitro FLS stimulation than MDMs, inferring differing roles in equine lung inflammation. Variability in recruitment and function of monocyte-macrophage populations warrant more detailed in vivo investigation in both homeostatic and diseased states. [ABSTRACT FROM AUTHOR]

Background: The intestinal tract is a rich and complex environment and its microbiota has been shown to have an important role in health and disease in the host. Several factors can cause disruption of the normal intestinal microbiota, including antimicrobial therapy, which is an important cause of diarrhea in horses. This study aimed to characterize changes in the fecal bacterial populations of healthy horses associated with the administration of frequently used antimicrobial drugs. Results: Twenty-four adult mares were assigned to receive procaine penicillin intramuscularly (IM), ceftiofur sodium IM, trimethoprim sulfadiazine (TMS) orally or to a control group. Treatment was given for 5 consecutive days and fecal samples were collected before drug administration (Day 1), at the end of treatment (Days 5), and on Days 14 and 30 of the trial. High throughput sequencing of the V4 region of the 16S rRNA gene was performed using an Illumina MiSeq sequencer. Significant changes of population structure and community membership were observed after the use of all drugs. TMS caused the most marked changes on fecal microbiota even at higher taxonomic levels including a significant decrease of richness and diversity. Those changes were mainly due to a drastic decrease of Verrucomicrobia, specifically the "5 genus incertae sedis". Changes in structure and membership caused by antimicrobial administration were specific for each drug and may be predictable. Twenty-five days after the end of treatment, bacterial profiles were more similar to pre-treatment patterns indicating a recovery from changes caused by antimicrobial administration, but differences were still evident, especially regarding community membership. Conclusions: The use of systemic antimicrobials leads to changes in the intestinal microbiota, with different and specific responses to different antimicrobials. All antimicrobials tested here had some impact on the microbiota, but TMS significantly reduced bacterial species richness and diversity and had the greatest apparent impact on population structure, specifically targeting members of the Verrucomicrobia phylum. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the fecal shedding of C. difficile in calves on farms in Sao Paulo State, Brazil. Fecal samples (n = 300) were collected from diarrheic (n = 78) and nondiarrheic (n = 222) calves less than 60 days of age from 20 farms. Fecal samples were inoculated into enrichment broth supplemented with taurocholate and cultured under anaerobic conditions. Colonies suspected to be C. difficile were harvested for DNA extraction and then multiplex PCR for the detection of genes encoding toxins A and B and binary toxins. All toxigenic isolates were ribotyped and tested for antimicrobial susceptibility, and five selected strains were subjected to whole-genome sequencing to determine their sequence type. C. difficile was isolated from 29.3 % (88/300) of the samples. All toxigenic isolates (17/88, 19.3 %) were classified as ribotypes RT046 (13/17–79.47 %, A+B+ CDT−) and RT126 (4/17 = 20.53 %, A+B+ CDT+). The sequenced strains from RT046 were classified as ST35 (Clade 1), while those from RT126 were classified as ST11 (Clade 5). No associations between the epidemiological factors in any of the groups and C. difficile isolation were observed. Most of the toxigenic isolates (16/17 = 94.41 %) were classified as multidrug-resistant. Calves can be an important source of toxigenic C. difficile strains, including multidrug-resistant isolates from ribotypes commonly observed in humans. • C. difficile was isolated from fecal samples of calves with and without diarrhea. • Toxigenic isolates were classified as RT046 and RT126. • Most of the toxigenic isolates were classified as multidrug resistant. [ABSTRACT FROM AUTHOR]

Objective--To determine effects of extracorporeal shock wave therapy (ESWT) on healing of wounds in the distal portion of the forelimb in horses. Animals-6 horses. Procedures--Five 6.25-cm² superficial wounds were created over both third metacarpi of 6 horses. Forelimbs were randomly assigned to treatment (ESWT and bandage) or control (bandage only) groups. In treated limbs, each wound was treated with 625 shock wave pulses from an unfocused electrohydraulic shock wave generator. In control limbs, each wound received sham treatment. Wound appearance was recorded weekly as inflamed or healthy and scored for the amount of protruding granulation tissue. Standardized digital photographs were used to determine the area of neoepithelialization and absolute wound area. Biopsy was performed on 1 wound on each limb every week for 6 weeks to evaluate epithelialization, fibroplasia, neovascularization, and inflammation. Immunohistochemical staining fora smooth muscle actin was used to label myofibroblasts. Results--Control wounds were 1.9 times as likely to appear inflamed, compared with treated wounds. Control wounds had significantly higher scores for exuberant granulation tissue. Treatment did not affect wound size or area of neoepithelialization. No significant difference was found for any of the histologic or immunohistochemical variables between groups. Conclusions and Clinical Relevance--Treatment with ESWT did not accelerate healing of equine distal limb wounds, but treated wounds had less exuberant granulation tissue and appeared healthier than controls. Therefore, ESWT may be useful to prevent exuberant granulation tissue formation and chronic inflammation of such wounds, but further studies are necessary before recommending ESWT for clinical application. [ABSTRACT FROM AUTHOR]

Introduction: In the equine racehorse industry, reduced athletic performance due to joint injury and lameness has been extensively reviewed. Intra-articular injections of glucocorticoids are routinely used to relieve pain and inflammation associated with osteoarthritis. Intra-articular injections of pharmaceutical agents require practice for precise needle placement and to minimize complications. Training on simulators or models is a viable alternative for developing these technical skills. The purpose of this study was to compare the qualitative ultrasonographic characteristics of three-dimensional (3D) printed models of equine cervical articular process joints to that of a dissected equine cervical spine (gold standard). Methods: A randomized complete block design study was conducted in which a total of thirteen cervical articular process joint models were printed using several materials, printers, and printing technologies. Ultrasound video clips with the models immersed in water were recorded. Two board certified veterinary radiologists and three veterinary radiology residents reviewed the videos and responded to a survey assessing and comparing the ultrasonographic characteristics of the 3D printed models to those of the gold standard. Results: Six 3D printed models had ultrasonographic characteristics similar to the gold standard. These six models were (material, printer, printing technology): nylon PA 12, EOS Formiga P100, selective laser sintering (P = 0.99); Onyx nylon with chopped carbon fiber, Markforged Onyx Two, fused deposition modeling (P = 0.48); polycarbonate, Ultimaker 3, fused deposition modeling (P = 0.28); gypsum, ProJet CJP 660 Pro, ColorJet Printing (P = 0.28); polylactic acid, Prusa I3, fused deposition modeling (P = 0.23); and high temperature V1 resin, Form 2, stereolithography (P = 0.22). Conclusion: When assessed in water, it is possible to replicate the qualitative ultrasonographic characteristics of bone using three dimensional printed models made by combining different materials, printing technologies, and printers. However, not all models share similar qualitative ultrasonographic characteristics with bone. We suggest that the aforementioned six models be used as proxy for simulating bones or joints for use with ultrasound. In order to replicate the resistance and acoustic window provided by soft tissues, further work testing the ability of these models to withstand embedding in material such as ballistic gelatin is required. [ABSTRACT FROM AUTHOR]

Objectives: To investigate whether lipopolysaccharide (LPS) is present in plasma of calves with naturally occurring diarrhea. The second objective was to determine whether plasma [LPS] correlates with clinical, hematological, biochemical, and acid‐base variables, and whether [LPS] differs between surviving and nonsurviving diarrheic calves. Design: Prospective observational study (January 2012–May 2014). Setting: Veterinary teaching hospital. Animals: Thirty‐four calves <28 days old admitted for diagnosis and treatment of diarrhea and 30 healthy control calves. Measurements and Main Results: Admission demographics, physical examination, blood gas, biochemistry analysis, and outcome data were recorded. Plasma concentration of LPS was determined using a bovine LPS ELISA assay. Plasma [LPS] was detected in both healthy and diarrheic calves. Plasma [LPS] was significantly higher in diarrheic than healthy calves (median: 0.99 ng/mL; Interquartile range (IQR): 0.068, vs 0.88 ng/mL; 0.065 ng/mL, respectively; P < 0.001). Plasma [LPS] was higher in nonsurviving (1.04 ng/mL; 0.07 ng/mL) than in surviving calves (0.98 ng/mL; 0.022 ng/mL; P < 0.001). Plasma [LPS] was higher in beef (1.07 ng/mL; 0.182 ng/mL) than in dairy diarrheic calves (0.99 ng/mL; 0.022 ng/mL; P < 0.001). In diarrheic calves, plasma [LPS] correlated with [l‐lactate] (r2 = 0.496; P = 0.002); hypoglycemia (r2 = −0.453; P = 0.007); increased unmeasured strong ions (r2 = 0.332; P = 0.050), [Mg2+] (r2 = 0.475; P = 0.004), and [phosphate] (r2 = 0.468; P = 0.005), and increased aspartate aminotransferase activity (r2 = 0.348; P = 0.003). Conclusions: This study highlights a potential role of LPS in the pathogenesis of metabolic derangements such as hyperlactatemia, hypoglycemia, and increased concentration of unmeasured strong anions in diarrheic calves. Further investigation evaluating the effect of LPS on l‐lactate and glucose metabolism in diarrheic calves is warranted. [ABSTRACT FROM AUTHOR]

Collagenous lectins are a family of soluble pattern recognition receptors that play an important role in innate immune resistance to infectious disease. Through recognition of carbohydrate motifs on the surface of pathogens, some collagenous lectins can activate the lectin pathway of complement, providing an effective means of host defense. Genetic polymorphisms in collagenous lectins have been shown in several species to predispose animals to a variety of infectious diseases. Infectious diseases are an important cause of morbidity in horses, however little is known regarding the role of equine collagenous lectins. Using a high-throughput, targeted re-sequencing approach, the relationship between genetic variation in equine collagenous lectin genes and susceptibility to disease was investigated. DNA was isolated from tissues obtained from horses submitted for post-mortem examination. Animals were divided into two populations, those with infectious or autoinflammatory diseases ( n  = 37) and those without ( n  = 52), and then subdivided by dominant pathological process for a total of 21 pools, each containing 4–5 horses. DNA was extracted from each horse and pooled in equimolar amounts, and the exons, introns, upstream (approximately 50 kb) and downstream (approximately 3 kb) regulatory regions for the 11 equine collagenous lectin genes and related MASP genes were targeted for re-sequencing. A custom target capture kit was used to prepare a sequencing library, which was sequenced on an Illumina MiSeq. After implementing quality control filters, 4559 variants were identified. Of these, 92 were present in the coding regions (43 missense, 1 nonsense, and 48 synonymous), 1414 in introns, 3029 in the upstream region, and 240 in the downstream region. In silico analysis of the missense short nucleotide variants identified 12 mutations with potential to disrupt collagenous lectin protein structure or function, 280 mutations located within predicted transcription factor binding sites, and 95 mutations located within predicted microRNA binding elements. Analysis of allelic association identified 113 mutations that segregated between the infectious/autoinflammatory and non-infectious populations. The variants discovered in this experiment represent potential genetic contributors to disease susceptibility of horses, and will serve as candidates for further population-level genotyping. This study contributes to the growing body of evidence that pooled, high-throughput sequencing is a viable strategy for cost-effective variant discovery. [ABSTRACT FROM AUTHOR]

This study compared the prevalence of C. innocuum DNA in the feces of healthy horses and horses with acute colitis. C. innocuum was identified in 22% (15/68) of colitis cases and 18% (12/68) of healthy horses (p = 0.416). • Clostridium innocuum prevalence in healthy and diarrheic horses is similar. • Clostridium innocuum detection is not associated with non-survival. • PCR can be used to detect Clostridium innocuum DNA in the feces of horses. [ABSTRACT FROM AUTHOR]

Neorickettsia (formerly Ehrlichia) risticii is an obligatory intracellular bacterium of digenetic trematodes. When a horse accidentally ingests aquatic insects containing encysted trematodes infected with N. risticii, the bacterium is transmitted from trematodes to horse cells and causes an acute and often fatal disease called Potomac horse fever (PHF). Since the discovery of N. risticii in the United States in 1984, using immunofluorescence and PCR assays, PHF has been increasingly recognized throughout North America and South America. However, so far, there exist only a few stable N. risticii culture isolates, all of which are from horses within the United States, and the strain diversity and environmental spreading and distribution of pathogenic N. risticii strains remain poorly understood. This paper reports the isolation of N. risticii from the blood of a horse with acute PHF in Ontario, Canada. Intracellular N. risticii colonies were detected in P388D1 cells after 47 days of culturing and 8 days after the addition of rapamycin. Molecular phylogenetic analysis based on amino acid sequences of major surface proteins P51 and Ssa1 showed that this isolate is distinct from any previously sequenced strains but closely related to midwestern U.S. strains. This is the first Canadian strain cultured, and a new method was developed to reactivate dormant N. risticii to improve culture isolation. [ABSTRACT FROM AUTHOR]

Clostridium difficile is a gram-positive, spore-forming enteric anaerobe which can infect humans and a wide variety of animal species. Recently, the incidence and severity of human C. difficile infection has markedly increased. In this study, we evaluated the genomic content of 73 C. difficile strains isolated from humans, horses, cattle, and pigs by comparative genomic hybridization with microarrays containing coding sequences from C. difficile strains 630 and QCD-32g58. The sequenced genome of C. difficile strain 630 was used as a reference to define a candidate core genome of C. difficile and to explore correlations between host origins and genetic diversity. Approximately 16% of the genes in strain 630 were highly conserved among all strains, representing the core complement of functional genes defining C. difficile. Absent or divergent genes in the tested strains were distributed across the entire C. difficile 630 genome and across all the predicted functional categories. Interestingly, certain genes were conserved among strains from a specific host species, but divergent in isolates with other host origins. This information provides insight into the genomic changes which might contribute to host adaptation. Due to a high degree of divergence among C. difficile strains, a core gene list from this study offers the first step toward the construction of diagnostic arrays for C. difficile. [ABSTRACT FROM AUTHOR]

We investigated Clostridium difficile in calves and the similarity between bovine and human C. difficile PCR ribotypes by conducting a case-control study of calves from 102 dairy farms in Canada. Fecal samples from 144 calves with diarrhea and 134 control calves were cultured for C. difficile and tested with an ELISA for C. difficile toxins A and B. C. difficile was isolated from 31 of 278 calves: 11 (7.6%) of 144 with diarrhea and 20 (14.9%) of 134 controls (p = 0.009). Toxins were detected in calf feces from 58 (56.8%) of 102 farms, 57 (39.6%) of 144 calves with diarrhea, and 28 (20.9%) of 134 controls (p = 0.0002). PCR ribotyping of 31 isolates showed 8 distinct patterns; 7 have been identified in humans, 2 of which have been associated with outbreaks of severe disease (PCR types 017 and 027). C. difficile may be associated with calf diarrhea, and cattle may be reservoirs of C. difficile for humans. [ABSTRACT FROM AUTHOR]