[Survey on the use of antibiotics among Swiss equine veterinarians]. / Umfrage zum Antibiotikagebrauch bei Schweizer Tierärzten mit Pferdeanteil.

INTRODUCTION: The aim of this study was to evaluate the current use of antibiotics by Swiss equine veterinarians and to compare the results with a similar study from 2013 before the introduction of the web tool Antibiotic Scout. The survey was sent to equine veterinarians according to the member database of the Swiss Veterinary Association (GST, SVS). The demographic data of the respondents and their antibiotics usage were collected. In addition, six different case scenarios were presented with questions to their potential antibiotic usage, active substance/preparation and the dosing scheme. The dosage provided was compared with the dosage information approved by Swissmedic in the information for healthcare professionals and the recommendations of the antibiotic scout. A backward logistic regression analysis was performed to assess the association between different aspects of antibiotic use and demographic data. The response rate was 94/739 (13 %), 22 of the 94 (23 %) had also participated in the 2013 study. 47/94 (50 %) of the respondents obtained their information from the antibiotic scout. The respondents indicated that they used an antibiotic in 16 %-88 % depending on the case scenario. Neither 3rd nor 4th generation cephalosporins or fluoroquinolones were used in the case scenarios. Dihydrostreptomycin was indicated as a possible antibiotic in a case scenario by 14/94 (15 %) of the respondents. Respondents who had already taken part in the 2013 survey used dihydrostreptomycin significantly more frequently (7/22, 32 % vs. 7/72, 10 %; p = 0,047). 29/81 (36 %) had underdosed compared to the prescribing information and 38/81 (47 %) compared to the antibiotic scout; neither was associated with demographic data. The use of non-equine-licensed antimicrobial products was directly related to the number of veterinarians in the practice (p = 0,007) and to the percentage of horses (p = 0,02). No association between demographics and peri-operative antibiotic use >24h (17/44, 39 %) was detected. The antibiotic prescribing habits of Swiss equine veterinarians have improved over the last 10 years. The antibiotic use decreased compared to the study of Schwechler et al. in 2013 by 0-16 % depending on the case scenario. The use of 3rd and 4th generation cephalosporins was reduced by 4 % and fluoroquinolones by 7 %. Underdosing according to scientific recommendations was reduced by 32 %. Furthermore, there is a need for additional information regarding the indication for antimicrobial use and the adequate use of perioperative antibiotics.

INTRODUCTION: L'objectif de cette étude était d'évaluer l'utilisation actuelle des antibiotiques par les vétérinaires équins suisses et de comparer les résultats avec une étude similaire de 2013 avant l'introduction de l'outil web AntibioticScout. L'enquête a été envoyée aux vétérinaires équins selon la base de données des membres de la Société des Vétérinaires Suisses (SVS). Les données démographiques des répondants et leur utilisation d'antibiotiques ont été collectées. En outre, six scénarios de cas différents ont été présentés avec des questions quant à l'utilisation potentielle d'antibiotiques, la substance active/préparation et le schéma posologique. La posologie fournie a été comparée à celle approuvée par Swissmedic dans les informations destinées aux professionnels de la santé et aux recommandations de l'AntibioticScout. Une analyse de régression logistique inverse a été réalisée pour évaluer l'association entre les différents aspects de l'utilisation des antibiotiques et les données démographiques. Le taux de réponse était de 94/739 (13 %), 22 des 94 (23 %) avaient également participé à l'étude de 2013. 47/94 (50 %) des répondants ont obtenu leurs informations à partir de AntibioticScout. Les répondants ont indiqué avoir utilisé un antibiotique dans 16 %-88 % selon le cas de figure. Ni les céphalosporines de 3ème ou 4ème génération ni les fluoroquinolones n'ont été utilisées dans les scénarios de cas. La dihydrostreptomycine a été indiquée comme un antibiotique possible dans un scénario de cas par 14/94 (15 %) des répondants. Les répondants qui avaient déjà participé à l>enquête de 2013 ont utilisé la dihydrostreptomycine significativement plus fréquemment (7/22, 32 % vs. 7/72, 10 % ; p = 0,047). 29/81 (36 %) avaient sous-dosé par rapport aux informations de prescription et 38/81 (47 %) par rapport à AntibioticScout ; ni l'un ni l'autre n'était associé aux données démographiques. L'utilisation de produits antimicrobiens non homologués pour les équidés était directement liée au nombre de vétérinaires dans le cabinet (p = 0,007) et au pourcentage de chevaux (p = 0,02). Aucune association entre les données démographiques et l'utilisation périopératoire d'antibiotiques >24h (17/44, 39 %) n'a été détectée. Les habitudes de prescription d'antibiotiques des vétérinaires équins suisses se sont améliorées au cours des 10 dernières années. L'utilisation d'antibiotiques a diminué par rapport à l'étude de Schwechler et al. en 2013 de 0 à 16 % selon le scénario du cas. L'utilisation des céphalosporines de 3ème et 4ème génération a été réduite de 4 % et les fluoroquinolones de 7 %. Le sous-dosage selon les recommandations scientifiques a été réduit de 32 %. Il est d'autre part nécessaire d>obtenir des informations supplémentaires concernant l>indication de l>utilisation d>antimicrobiens et l>utilisation adéquate des antibiotiques périopératoires.

A case series highlighting the role of different gamma-herpesviruses in Equine Multinodular Pulmonary Fibrosis.

INTRODUCTION: This case series describes three cases of equine multinodular pulmonary fibrosis (EMPF) diagnosed at the Clinic for Equine Internal Medicine at the University of Zurich between 2012 and 2017. Current information on etiology and treatment options are presented. Two horses showed mild signs of chronic lower respiratory tract disease and one horse was presented with acute signs of disease including recurrent fever spikes and tachypnea. Diagnosis was achieved by physical examination, radiographic findings, and PCR testing for equine herpesviruses (EHV) of bronchoalveolar lavage (BAL) fluid or lung tissue obtained by biopsy. All horses were euthanized due to continuing deterioration after attempted treatment. Post mortem histological examination of lung tissue showed severe multifocal diffuse to confluent fibrosis in two cases and in another horse a discrete nodular fibrosis pattern. Panherpes nested PCR revealed the presence of equine herpesvirus 5 (EHV-5) DNA in lung tissue of one horse whereas in two other horses, asinine herpes virus 5 (AHV-5) was detected. EMPF should be considered as a differential diagnosis in horses with acute and chronic respiratory disease, including horses non-responsive to treatment for equine asthma.

INTRODUCTION: Cette série de cas décrit trois cas de fibrose pulmonaire multinodulaire équine (EMPF) diagnostiqués à la Clinique de médecine interne équine de l'Université de Zurich entre 2012 et 2017. Des informations actuelles sur l'étiologie et les options de traitement sont présentées. Deux chevaux présentaient de légers signes de maladie chronique des voies respiratoires inférieures et un cheval présentait des signes aigus de maladie, notamment des pics de fièvre récurrents et une tachypnée. Le diagnostic a été obtenu grâce à un examen physique, des résultats radiographiques et des tests PCR pour les virus herpès équins (EHV) du liquide de lavage broncho-alvéolaire (BAL) ou du tissu pulmonaire obtenus par biopsie. Tous les chevaux ont été euthanasiés en raison d'une détérioration continue après une tentative de traitement. L'examen histologique post mortem du tissu pulmonaire a montré une fibrose multifocale diffuse à confluente sévère dans deux cas et chez un cheval un type de fibrose nodulaire discret. La PCR par Panherpes a révélé la présence d'ADN de virus herpès équin 5 (EHV-5) dans le tissu pulmonaire d>un cheval alors que chez deux autres chevaux, le virus de l>herpès asinien 5 (AHV-5) a été détecté. L'EMPF doit être considéré comme un diagnostic différentiel chez les chevaux souffrant d'une maladie respiratoire aiguë et chronique, y compris les chevaux ne répondant pas au traitement de l'asthme équin.

The effect of different antimicrobial treatment regimens on the faecal shedding of ESBL-producing Escherichia coli in horses.

Fourth-generation cephalosporins can select for extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae in horses, but it is unknown to what extent this occurs compared to penicillin/gentamicin combination treatment. The objective was to evaluate the effect of different antimicrobial treatments on faecal shedding and diversity of ESBL-producing Escherichia coli (ESBL-EC) in horses. Upon hospital admission, 86 horses in need of antimicrobial treatment or prophylaxis were randomly allocated to receive penicillin and gentamicin (PG) or cefquinome (CEF). Untreated horses were included as controls (NOAMD, n = 33). Faecal samples from admission (T1), 3 days after admission (T2), and faecal swabs 28 days after discharge (T3) were cultured selectively. Differences in prevalence (T1, T2, T3) and counts (T1, T2) of ESBL-EC between groups and over time were analysed. On a subset of ESBL-EC isolates, antimicrobial susceptibility testing (n = 45) and whole-genome sequencing followed by SNP-analysis (n = 46) were performed. The prevalence of ESBL-EC at T1 was 12 % with no significant difference between groups. In all groups, significantly higher carriage rates were observed at T2 and T3 compared to T1. Carriage and counts of ESBL-EC at T2 were significantly higher in treated compared to untreated horses. There was no significant difference between PG and CEF at any time points. Despite a large genetic diversity, indistinguishable ESBL clones were observed in different horses over time. In conclusion, antimicrobial treatment and hospitalization increased prevalence and counts of ESBL-EC, and transmission of ESBL-EC in the hospital was suspected. These findings highlight the importance of antimicrobial stewardship and infection control practices in equine medicine.

Prevalence of Clostridium difficile and Clostridium perfringens in Swiss horses with and without gastrointestinal disease and microbiota composition in relation to Clostridium difficile shedding.

Overgrowth of enteric clostridia in dysbiosis in horses with colic is presumed but scarcely investigated. The objective was to provide prevalence data of Clostridium difficile and Clostridium perfringens in horses with and without gastrointestinal disease in Switzerland, and investigate microbiota differences between C. difficile shedders and non-shedders. Fecal samples were taken from healthy horses (n = 103), horses with colic (n = 98) and horses with diarrhea (n = 151). Colic horses were sampled on three days. Selective enrichment culture and molecular typing for C. difficile and C. perfringens was performed. Microbiota differences between horses with colic shedding (n = 7) and not shedding (n = 7) C. difficile were assessed using metagenomic sequencing. The cumulative prevalence (19% C. difficile; 16% C. perfringens) was higher compared to single day samples (1-10% C. difficile; 3-8% C. perfringens, all p < 0.003). Horses with colic shed significantly more C. difficile (p < 0.001) but not C. perfringens (p = 0.09) compared to healthy horses. Prevalence in horses with diarrhea was 8% for both Clostridium species. There were no significant microbiota differences between C. difficile shedders and non-shedders with regards to relative abundance on any phylogenetic level, and alpha diversity. Limited differences were seen on LEfSE analysis and in beta diversity indices. Multiple fecal samples should be taken when investigating shedding of enteric clostridia. As horses with colic shed more enteric clostridia compared to healthy horses special biosecurity protocols for horses with colic should be considered in hospitals. Differences in microbiota composition between C. difficile shedders and non-shedders were limited. Further studies on the role of dysbiosis in C. difficile are needed.

Complications of intravenous catheterization in horses.

INTRODUCTION: Intravenous catheterization is a necessity for continuous administration of intravenous fluids and for intermittent intravenous access to avoid discomfort and potential complications of repeated needle insertions into the vein. Intravenous catheterization is commonly performed and well tolerated in horses, but catheter associated complications have been reported. The most commonly reported complication is thrombophlebitis, but others such as venous air embolism, exsanguination and catheter fragmentation may also occur. This article aims to review clinical signs, pathogenesis, diagnosis, therapy, risk factors and prevention of common catheter associated complications.

INTRODUCTION: Le cathétérisme veineux est une nécessité pour l'administration continue de fluides par voie intraveineuse et pour garantir un accès veineux intermittent afin d'éviter l'inconfort et les complications potentielles liées à la pénétration répétée d'une aiguille dans la veine. Le cathétérisme veineux est usuellement pratiqué chez le cheval et il est bien toléré mais des complications associées sont rapportées. La plus commune d'entre elle est la thrombophlébite mais d'autre, telles l'embolie gazeuse, l'exsanguination ou la fragmentation du cathéter peuvent aussi survenir. Cet article vise à résumer les signes cliniques, la pathogénèse, le diagnostic, le traitement les facteurs de risque et la prévention des complications communément associées avec le cathétérisme.

Validation of a Point-of-Care Quantitative Equine IgG Turbidimetric Immunoassay and Comparison of IgG Concentrations Measured with Radial Immunodiffusion and a Point-of-Care IgG ELISA.

BACKGROUND: Point-of-care (POC) diagnostic tests with good sensitivity and specificity are needed for diagnosing failure of transfer of passive immunity (FTPI) in foals. Turbidimetric immunoassays (TIA) have these characteristics and provide quantitative results. A commercially available TIA-based POC test (POC-TIA) has not been validated in horses. OBJECTIVE: To validate a POC-TIA and compare results of POC-TIA, a POC-ELISA, and radial immunodiffusion (RID). ANIMALS: Heparinized blood samples (n = 127) from 48 hospitalized foals (<12 hour to 48 days). METHODS: Prospective validation study. IgG concentrations were measured using RID (gold standard), POC-TIA, and POC-ELISA. Agreement between assays was assessed using Bland-Altman analysis. Sensitivity and specificity were calculated using ROC curves. Inter- and intra-assay coefficients of variation (CVs) and linearity were evaluated for POC-TIA. RESULTS: The mean bias (95% limits of agreement) between RID and POC-TIA was -4 (-185 to 176), 27 (-201 to 255), and 308 (-377 to 993) mg/dL for samples with IgG concentrations of <400, 400-800, and >800 mg/dL, respectively. Sensitivity and specificity at optimal cutoff were 94 and 100% for the POC-TIA and 94 and 100% for the POC-ELISA to detect IgG <400 mg/dL, and 85 and 87% (POC-TIA) and 69 and 79% (POC-ELISA) to detect IgG ≤800 mg/dL. Intra- and interassay CVs for POC-TIA ranged between 1.6-3.8 and 11.9-18.8%, respectively. Linearity of the dilution series was preserved (R2 > 0.96). CONCLUSIONS AND CLINICAL IMPORTANCE: The POC-TIA provided unambiguous results and had sufficient sensitivity, specificity, accuracy, and precision to be used as an alternative to other POC tests to assess FTPI in foals.

Comparison of the fecal bacterial microbiota of healthy and diarrheic foals at two and four weeks of life.

BACKGROUND: Diarrhea in foals affects up to 60% of foals during the first six months of life. The effect of diarrhea on the fecal bacterial microbiota in foals has not been investigated. Little is known on the fecal bacterial microbial richness and diversity of foals at a young age. The objective was to compare the fecal bacterial microbiota of healthy foals to foals with diarrhea at two and four weeks of life. METHODS: Fecal samples were collected from foals (n = 20) at 1-14 (T1) and 15-28 (T2) days of age and analyzed using high throughput sequencing. Differences in relative abundance of bacterial taxa, alpha diversity and beta diversity indices were assessed between age-matched foals with diarrhea (n = 9) and healthy foals (n = 11), and between time points. RESULTS: Differences in microbial community composition based on time point and health status were observed on all taxonomic levels. Of 117 enriched species in healthy foals at T2, 50 (48%) were Lachnospiraceae or Ruminococcaceae. The Chao richness index was increased in healthy foals at T2 compared to T1 (p = 0.02). Foals with diarrhea had a significantly lower richness index than non-diarrheic foals at T2 (p = 0.04). Diarrhea had an inconsistent effect, while time point had a consistent effect on microbial community structure. CONCLUSIONS: Preventative and therapeutic measures for diarrhea should focus on maintaining bacterial microbiota richness. Lachnospiraceae and Ruminococcaceae were underrepresented in foals with diarrhea. These should be evaluated further as potential therapeutic options.

A retrospective analysis of antimicrobial resistance in bacterial pathogens in an equine hospital (2012-2015).

INTRODUCTION: Antimicrobial resistance has become an important concern in veterinary medicine. The aim of this study was to describe the rate of antimicrobial resistance in common equine pathogens and to determine the occurrence of multidrug-resistant isolates. A retrospective analysis of all susceptibility testing results from bacterial pathogens cultured from horses at the University of Zurich Equine Hospital (2012-2015) was performed. Strains exhibiting resistance to 3 or more antimicrobial categories were defined as multidrug-resistant. Susceptibility results from 303 bacterial pathogens were analyzed, most commonly Escherichia coli (60/303, 20%) and Staphylococcus aureus (40/303, 13%). High rates of acquired resistance against commonly used antimicrobials were found in most of the frequently isolated equine pathogens. The highest rate of multidrug resistance was found in isolates of Acinetobacter baumannii (23/24, 96%), followed by Enterobacter cloacae complex (24/28, 86%) and Escherichia coli (48/60, 80%). Overall, 60% of Escherichia coli isolates were phenotypically ESBL-producing and 68% of Staphylococcus spp. were phenotypically methicillin-resistant. High rates of acquired antimicrobial resistance towards commonly used antibiotics are concerning and underline the importance of individual bacteriological and antimicrobial susceptibility testing to guide antimicrobial therapy. Minimizing and optimizing antimicrobial therapy in horses is needed.

INTRODUCTION: Une connaissance des bactéries pathogènes présentes localement est importante pour pouvoir mettre en œuvre dès le début un traitement antibiotique ciblé. Le but de cette étude était de décrire les agents infectieux les plus fréquents et leur évolution dans le temps chez les chevaux hospitalisés à l'hôpital vétérinaire de Zürich entre 1988 et 2014. Tous les résultats positifs de cultures ont été analysés rétrospectivement. Les isolats ont été classés sur la base de leur localisation, du diagnostic et de l'année de leur isolation. Diverses périodes ont été comparées au moyen d'un test de chi carré. Escherichia coli (299/1'723, 17%), Streptococcus equi ssp. zooepidemicus (295/1'723, 17%) et les staphylocoques coagulase positive (196/1'723, 11%) ont été dans l'ensemble les bactéries mises en évidence le plus souvent. Des bactéries anaérobes strictes ont été trouvées principalement dans les infections dentaires (18/66, 27%), les péritonites (6/43, 14%) et les infections des tissus mous (28/208, 13%). Des cultures mixtes étaient particulièrement fréquentes dans les infections cutanées et dentaires (24/55, 44% et 17/43, 40%). Les principaux germes isolés dans la plupart des systèmes d'organes étaient aussi bien Gram positifs que Gram négatifs. Pour cette raison, il conviendrait, jusqu'à ce que les résultats des cultures et des antibiogrammes soient connus, de choisir un médicament à large spectre d'action.

The longitudinal effect of a multi-strain probiotic on the intestinal bacterial microbiota of neonatal foals.

REASONS FOR PERFORMING THE STUDY: The microbiota plays a key role in health and disease. Probiotics are a potential way to therapeutically modify the intestinal microbiota and prevent disease. OBJECTIVES: The aim of this study was to investigate the effects of probiotics on the bacterial microbiota of foals during and after administration. STUDY DESIGN: Randomised placebo controlled field trial. METHODS: Thirty-eight healthy neonatal foals enrolled in a previous study were selected. The foals had received a multi-strain probiotic (four Lactobacillus spp. 3-4 × 103 colony-forming units (cfu)/g each, Bifidobacterium animalis spp. lactis, 1 × 103-4 cfu/g) or placebo once daily for 3 weeks. A total of 3 faecal samples were collected from each foal at 2-week intervals and assessed via metagenomic sequencing. The Wilcoxon test was used to compare data between treatment groups. RESULTS: There were no changes on the phylum, order or class level between treatment groups at any age (all P>0.05) but some significant changes in relative abundance of families. Probiotic administration did not result in an increased relative abundance of lactobacilli or bifidobacteria at any age (Lactobacillus: P = 0.9, P = 0.1 and P = 0.2, Bifidobacterium: P = 0.3, P = 0.6 and P = 0.1 for Weeks 2, 4 and 6, respectively). Lactobacillus was enriched in the probiotic group at Week 6 on LEfSe analysis (linear discriminant analysis score 0.34, P = 0 .02). There was no effect on alpha diversity (all P>0.2) or community structure when parsimony and unifrac analysis were applied (all P>0.6). CONCLUSIONS: There were limited effects of probiotic treatment on the bacterial microbiota of foals. The studied probiotic based on lactobacilli and bifidobacteria has a limited potential for therapeutic modification of the gastrointestinal microbiota.

Effects of transport, fasting and anaesthesia on the faecal microbiota of healthy adult horses.

REASONS FOR PERFORMING STUDY: The intestinal microbiota is important for health and disease. Factors that disturb the equine intestinal microbiota need further investigation. OBJECTIVES: To determine the effects of transport, fasting and anaesthesia on the faecal microbiota of healthy adult horses using next-generation sequencing. STUDY DESIGN: Experimental trial. METHODS: Faecal samples were taken from 8 horses at baseline, after transport, 12 h of fasting and 24, 48 and 72 h after a 6 h anaesthesia. Next generation sequencing of the V4 region of the 16S rRNA gene was used to assess the microbial composition of faeces. Alpha diversity, phylogenetic structures and beta diversity were assessed. RESULTS: There were significant changes in the relative abundances of phyla, classes, orders and families after transport, fasting and anaesthesia. Most notably horses had a significantly lower abundance of Clostridiales after transport compared with baseline (P = 0.03) and a decreased abundance of Rickettsiales after fasting (P = 0.024). Alpha diversity was not significantly different between time points (all P>0.21). When parsimony analysis was applied, anaesthesia had a significant effect on community membership and structure (Jaccard index and Yue and Clayton index both P = 0.02). CONCLUSIONS: There was some effect of transport, fasting and anaesthesia on the composition and structure of the microbiota of healthy horses. This indicates these are potentially stress factors for the equine intestinal microbiota. Further investigation is required to look at the potential impact of changes in the microbiota on the development of disease in the post anaesthetic period.

The Use of Recombinant Tissue Plasminogen Activator (rTPA) in The Treatment of Fibrinous Pleuropneumonia in Horses: 25 Cases (2007-2012).

BACKGROUND: Information about treatment protocols, adverse effects and outcomes with intrapleural recombinant tissue plasminogen activator (rTPA) use in horses with fibrinous pleuropneumonia is limited. HYPOTHESIS/OBJECTIVES: Describe factors that contribute to clinical response and survival of horses treated with rTPA intrapleurally. ANIMALS: Horses with bacterial pneumonia and fibrinous pleural effusion diagnosed by ultrasonography, that were treated with rTPA intrapleurally. METHODS: Retrospective multicenter case series from 2007-2012. Signalment, history, clinical and laboratory evaluation, treatment, and outcome obtained from medical records. Regression analysis used to identify associations between treatments and outcomes. RESULTS: Thirty three hemithoraces were treated in 25 horses, with 55 separate treatments. Recombinant tissue plasminogen activator (375-20,000 µg/hemithorax) was administered 1-4 times. Sonographically visible reduction in fibrin mat thickness, loculations, fluid depth, or some combination of these was seen in 32/49 (65%) treatments. Response to at least 1 treatment was seen in 17/20 (85%) horses with sonographic follow-up evaluation after every treatment. Earlier onset of rTPA treatment associated with increased survival odds. No association was found between cumulative rTPA dose or number of rTPA doses and survival, development of complications, duration of hospitalization or total charges. Clinical evidence of hypocoagulability or bleeding was not observed. Eighteen horses (72%) survived to discharge. CONCLUSIONS AND CLINICAL IMPORTANCE: Treatment with rTPA appeared safe and resulted in variable changes in fibrin quantity and organization within the pleural space. Recombinant tissue plasminogen activator could be a useful adjunct to standard treatment of fibrinous pleuropneumonia, but optimal case selection and dosing regimen remain to be elucidated.

Effect of a probiotic on prevention of diarrhea and Clostridium difficile and Clostridium perfringens shedding in foals.

BACKGROUND: Up to 60% of foals develop diarrhea within 6 months after birth. Preventive measures are limited but potentially probiotics could be used. OBJECTIVE: To evaluate the effect of a newly designed probiotic on the incidence of foal diarrhea in a randomized field trial. ANIMALS: Seventy-two healthy neonatal foals. METHODS: Randomized, placebo-controlled field trial. Foals were administered a placebo or probiotic for 3 weeks and monitored for an additional week. A total of 3 fecal samples were taken from each foal at biweekly intervals. Statistical modeling was applied for comparison of incidence and duration of diarrhea and fecal shedding of Clostridium perfringens and Clostridium difficile between treatment and age groups. RESULTS: The overall incidence of diarrhea was 41 of 72 (59%) and did not differ (P = 0.37) between treatment groups. Foals treated with probiotics were more likely to develop diarrhea requiring veterinary intervention (P = 0.007). Age had a significant effect on incidence of diarrhea (P < 0.001); foals 8-15 days old having the highest probability of developing diarrhea. Duration of diarrhea and soft feces were not significantly different between groups. The prevalence of C. perfringens shedding was 55% with no difference between treatment groups (P = 0.23). The prevalence of C. difficile shedding was 11%. CONCLUSION AND CLINICAL IMPORTANCE: There was no benefit of administering a 3-week course of probiotics, but potential adverse effects were noted. Whether the probiotics lacked a clinical effect, or the choice of strains or dose was inadequate, is unknown. Clostridial shedding was not influenced by probiotics despite in vitro activity of probiotics.

Preliminary investigation of the area under the L-lactate concentration-time curve (LACArea) in critically ill equine neonates.

BACKGROUND: A variety of measures of L-lactate concentration ([LAC]) in the blood of critically ill neonatal foals have shown utility as prognostic indicators. These measures, evaluating either the severity of hyperlactatemia or the duration of exposure to hyperlactatemia, perform fairly well and have correctly classified 75-80% of foals examined in several studies. The area under the L-lactate concentration versus time curve (LACArea) encompasses both severity and duration of hyperlactatemia and should improve correct classification of patient survival. HYPOTHESIS/OBJECTIVES: LACArea is larger in nonsurviving critically ill neonatal foals. ANIMALS: Forty-nine foals admitted for critical illness to 1 of 4 referral hospitals. METHODS: Whole blood was obtained at admission and 6, 12, 18, and 24 hours after admission for measurement of L-lactate using a handheld lactate meter. LACArea was calculated for: admission-6, 6-12, 12-18, 18-24 hours, and admission-24 hours using the trapezoidal method and summing the 6-hours interval areas to determine total 24 hours area. Differences between survivors and nonsurvivors were determined using robust regression and Kruskal-Wallis testing, P < .05. RESULTS: LACArea was significantly larger in nonsurviving foals (n = 9) than in surviving foals (n = 40) at all time periods examined. CONCLUSIONS AND CLINICAL IMPORTANCE: Differences in LACArea between surviving and nonsurviving critically ill neonatal foals are large and support further investigation of this method as an improved biomarker for survival in critically ill neonatal foals is indicated.

Probiotic use in horses - what is the evidence for their clinical efficacy?

The gastrointestinal microbiota is extremely important for human and animal health. Investigations into the composition of the microbiota and its therapeutic modification have received increasing interest in human and veterinary medicine. Probiotics are a way of modifying the microbiota and have been tested to prevent and treat diseases. Probiotics are proposed to exert their beneficial effects through various pathways. Production of antimicrobial compounds targeting intestinal pathogens, general immune stimulation, and colonization resistance are among these mechanisms. Despite widespread availability and use, scientific, peer-reviewed evidence behind commercial probiotic formulations in horses is limited. Additionally, quality control of commercial over-the-counter products is not tightly regulated. Although promising in vitro results have been achieved, in vivo health benefits have been more difficult to prove. Whether the ambiguous results are caused by strain selection, dosage selection or true lack of efficacy remains to be answered. Although these limitations exist, probiotics are increasingly used because of their lack of severe adverse effects, ease of administration, and low cost. This review summarizes the current evidence for probiotic use in equine medicine. It aims to provide veterinarians with evidence-based information on when and why probiotics are indicated for prevention or treatment of gastrointestinal disease in horses. The review also outlines the current state of knowledge on the equine microbiota and the potential of fecal microbial transplantation, as they relate to the topic of probiotics.

In vitro inhibition of Clostridium difficile and Clostridium perfringens by commercial probiotic strains.

Probiotics have gained importance in human and veterinary medicine to prevent and control clostridial enteric disease. Limited information is available on the ability of different probiotic bacteria used in food products to inhibit Clostridium difficile and Clostridium perfringens. The objective of this study was to examine the in vitro inhibitory effects of selected commercial bacterial strains on pathogenic clostridia and their growth characteristics under simulated gastrointestinal conditions. The inhibitory effects of 17 commercial strains of Lactobacillus (n = 16) and Bifidobacterium (n = 1) on the reference strains of C. difficile and C. perfringens were assessed by an agar well diffusion assay and by a broth culture inhibition assay using cell-free supernatant harvested at different growth phases, with and without pH neutralization. To study growth characteristics, probiotic strains were cultivated in different acid and bile environments, and growth in the modified media was compared to growth in standard medium. In the agar well diffusion assay, supernatant obtained from two probiotic strains inhibited the growth of both reference and clinical strains of C. perfringens. This effect as seen when supernatant was assessed with and without pH neutralization. Supernatants obtained from 10 probiotic strains inhibited C. difficile only when supernatant was added without pH neutralization. In the broth culture inhibition assay, growth of C. perfringens and C. difficile was inhibited by supernatant without pH neutralization from 5 and 10 probiotic strains, respectively. All potential probiotic strains were able to grow at pH 4.0 and in the presence of 0.15% and 0.3% bile but none were able to grow or survive at pH 2.0. Altogether five probiotic strains [Lactobacillus plantarum (n = 2), Lactobacillus rhamnosus (n = 2), Bifidobacterium animalis lactis (n = 1)] were shown to inhibit all strains of C. difficile and C. perfringens. The inhibitory effect was probiotic strain-specific. Two strains showed a pH-independent inhibitory effect likely due to production of either antibiotics or bacteriocins inhibiting C. perfringens only. These strains have favourable growth characteristics for use as probiotics and their efficacy as prophylactic or therapeutic measures against clostridial enteric disease should be further evaluated by clinical trials in animals.

Longitudinal study of Clostridium difficile and antimicrobial susceptibility of Escherichia coli in healthy horses in a community setting.

Point prevalence studies have reported carriage rates of enteric pathogens in healthy horses, but longitudinal data are lacking. Commensal E. coli is an indicator organism to evaluate antimicrobial resistance of enteric bacteria, yet there are limited data for horses. The objectives of this study were to investigate and molecularly characterize isolates of Clostridium difficile, Clostridium perfringens and Salmonella, collected sequentially over a one year period, and to determine the antibiotic susceptibility profile for E. coli. Fecal samples were collected monthly from 25 adult horses for one year. Selective cultures were performed for all above bacteria. C. difficile isolates were characterized via PCR toxin gene profiling and ribotyping. Broth microdilution was performed to assess antimicrobial susceptibility profiles of E. coli. Toxigenic Clostridium difficile was isolated from 15/275 (5.45%) samples from 10/25 (40%) horses. Four horses were positive at multiple sampling times but different ribotypes were found in three. Ribotypes included 078 (n=6), 001 (n=6) and C (n=3). C. perfringens was not isolated, nor was Salmonella. E. coli was isolated from 232/300 (77%) fecal samples. Resistance to ≥ 1 and ≥ 3 antimicrobials was present in 31/232 (13.4%) and 6/232 (2.6%) respectively. Only two horses shed the same strain of toxigenic C. difficile for more than one month, indicating that shedding is transient. The high number of ribotype 078 is consistent with recent emergence of this strain in the local horse population. The low prevalence of antibiotic resistance in commensal E. coli suggests that healthy horses are not likely a major reservoir of resistance for enteric bacteria.

Consistent detection of bovine papillomavirus in lesions, intact skin and peripheral blood mononuclear cells of horses affected by hoof canker.

REASONS FOR PERFORMING THE STUDY: Equine hoof canker is a chronic proliferative pododermatitis of as yet unknown aetiology. Like equine sarcoid disease, canker is a therapy-resistant disorder characterised by hyperkeratosis, acanthosis and a marked tendency to recur. HYPOTHESIS: There is an association of sarcoid-inducing bovine papillomaviruses of types 1 and 2 (BPV-1, BPV-2) with hoof canker disease. METHODS: Using PCR-based techniques, we assessed canker tissue, intact skin and/or peripheral blood mononuclear cells (PBMCs) of 25 canker-affected horses for the presence of sarcoid-associated BPV-1 and -2. RESULTS: Conventional PCR revealed BPV-1/-2 DNA in 24/24 canker, 12/13 skin and 10/11 PBMC DNA isolates. Using inverse PCR, full-length BPV episomes were detected in 1/5 canker specimens. Sequencing of viral early and late genes amplified from canker, intact skin and PBMC DNA of 2 cases revealed an overall identity of 98% to BPV-1. Viral DNA loads amounted to ≤16 copies per cell in canker tissue and intact skin, and to ≤0.35 copies per PBMC, as determined by quantitative PCR. Using RT-PCR, the viral major oncogene E5 was shown to be transcribed in 2/4 canker tissue specimens and 5/7 PBMC isolates. Immunocapture PCR from 7 canker and 6 skin extract supernatants revealed capsomere-associated viral DNA in one canker and one skin sample. Hoof tissue, skin and PBMCs collected from 13 individuals with no signs of canker or BPV-related malignancies scored negative throughout the experiments. CONCLUSION: These findings suggest that the observed presence of BPV-1/-2 in canker-affected horses is not coincidental but indicative of an active contribution to hoof canker disease. POTENTIAL RELEVANCE: The use of antivirals and/or immune modulators may help improving canker therapy.