Using Viscum album extracts (ISCADOR) for successful management of neoplasms of the skin in horses and cats in consideration of aspects relevant to human medicine.

Background: Mistletoe extracts (Viscum album extracts, VAE) such as ISCADOR, have been used successfully in human oncology for many years. In veterinary medicine, there have been reports of successful treatment, but scientific data on efficacy are lacking. Two studies were therefore conducted to assess whether VAE can serve as an adjunct to standard treatment in horses and cats. Methods: As part of a placebo-controlled double-blind study (study 1), 53 horses with equine sarcoid (a solitary or multifocal skin tumor) were treated with VAE pini (pine mistletoe) in progressively increasing doses (n=32) or with a sodium chloride solution (n=21). Three times a week for 15 weeks. All horses were observed over a period of 12 months for tumor recurrence and cure. In an observational study (study 2), 44 cats with fibrosarcoma, a very common skin tumor with a high rate of recurrence, were treated postoperatively with 0.1% (0.5ml per dose) oral VAE quercus (oak mistletoe) twice daily, and disease free survival was measured. Results: In study 1, treatment with ISCADOR led to significantly better results than placebo. Improvement was seen in 41% of the cases (placebo 14%), and cure was achieved in 28% of the cases (placebo 14%). In study 2, disease-free survival time in cats treated postoperatively with VAE quercus was 438 days compared with 365 to 475 days for conventional chemotherapies and 120 to 261 days for surgery alone, as seen in literature. Aggressive surgery combined with radiation and chemotherapy led to markedly superior results (661 to 986 days). Conclusion: The study 1 was able to demonstrate, for the first time, the efficacy of VAE compared to placebo control. This study also showed that the effect of VAE apparently extends considerably beyond the treatment period; unexpectedly, only very few recurrences were observed between the end of treatment and the 12th month of observation. The results of the study 2 show that oral administration of mistletoe preparations also yields satisfactory prophylactic effects. The discussion surrounding oral treatment and the question whether it can replace injection treatment must therefore be continued, even in human medicine

Clinical Presentation and Laboratory Diagnostic Work-Up of a Horse with Tick-Borne Encephalitis in Switzerland

Tick-borne encephalitis is an important viral tick-borne zoonosis in Europe and Asia. The disease is induced by tick-borne encephalitis virus (TBEV). This report describes a 16-year-old Warmblood gelding presenting with sudden onset of lethargy, ataxia, and muscle fasciculations on the nostrils, the lips, and the eye lids as the most important clinical findings. The horse further had a mild facial nerve paralysis with drooping of the right upper and lower lip. Diagnosis was based on paired serum samples using TBEV-ELISAs revealing high serum IgM in the first sample with normal IgM in the second sample and an increase in serum IgG and neutralizing antibodies, indicating acute and recent infection. TBEV was confirmed by a virus-neutralization test, revealing a fivefold increase in antibodies 32 days after of the onset of clinical signs. Although the specific PCR on cerebrospinal fluid (CSF) was negative, TBEV-specific IgG and IgM were identified in the CSF of the horse. Treatment consisted of anti-inflammatory and anti-oxidative treatment and the horse recovered with a mild drooping of the right nostril as the only remaining clinical sign. TBEV infection is a potential differential diagnosis of neurological disease in horses living in endemic areas and this is the first report to describe the diagnostic criteria in a horse as recommended in humans with suspected TBEV infection

Antimicrobial susceptibility patterns of blood culture isolates from foals in Switzerland.

INTRODUCTION We report blood culture results of 43 foals admitted to an equine hospital for medical or surgical disorders and determine minimal inhibitory concentrations (MIC) of different antibiotics. Eleven foals had a positive blood culture result despite prior administration of antibiotics in 10 of these animals. MIC values above EUCAST and/or CLSI breakpoints were identified in coagulase-negative staphylococci, methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecium. Gram-negative isolates were less frequently identified and did not appear to exhibit increased MIC values. This study shows that bloodstream infections in foals in Switzerland are caused by diverse bacteria including Gram-positive bacteria which exhibit resistance to several classes of antibiotics

Long-Read Transcriptome of Equine Bronchoalveolar Cells

We used Pacific Biosciences long-read isoform sequencing to generate full-length transcript sequences in equine bronchoalveolar lavage fluid (BALF) cells. Our dataset consisted of 313,563 HiFi reads comprising 805 Mb of polished sequence information. The resulting equine BALF transcriptome consisted of 14,234 full-length transcript isoforms originating from 7017 unique genes. These genes consisted of 6880 previously annotated genes and 137 novel genes. We identified 3428 novel transcripts in addition to 10,806 previously known transcripts. These included transcripts absent from existing genome annotations, transcripts mapping to putative novel (unannotated) genes and fusion transcripts incorporating exons from multiple genes. We provide transcript-level data for equine BALF cells as a resource to the scientific community

Safety and Efficacy of Mistletoe extract (Viscum album) Iscador(R) P in Equine Sarcoid

Equine Sarcoid (ES), a semi malignant skin tumour, is the most common neoplasia of horses (Marti et al.1993). There is no generally effective single treatment existing and the recurrence rate is high (Martens et al.2001). Furthermore as a treatment obstacle, Sarcoids tend to occur as multi-focal tumours or can be found in critical locations (peri-ocular) where success of surgical interventions is very limited (Carstanjen et al.1997). Due to encouraging results in human medicine and small animals practice after adjuvant cancer treatment by mistletoe extracts (Biegel et al.2007) the viscum therapy could hypothetically fill a therapy gap for equine tumour diseases as well. Hence, goal of this clinical trial was to evaluate the safety and efficacy of Viscum album pini (Iscador® P) as a primary therapy of Equine Sarcoid in the context of a prospective, randomized, controlled, double-blind study. Material and Methods: 53 horses with histologically confirmed diagnosis Equine Sarcoid were randomly assigned as Iscador (I; n=32) or Control group (C; n=21). There are 23 mares, 25 geldings and 5 stallions ranged in age from 3 to 17 years with an average of 7.2 years. The horses received scalating dosages of Iscador® P starting at 0.1mg up to 20 mg extract per 1 ml dose or physiological NaCl solution 3 times a week over 105 days sub-cutanously into the pectoral region. The number, localization and morphology of the ES were observed and documented over 12 months. As target variable the positive outcome rate (PO rate) was calculated representing the patients status at the end of the investigation. Results: In the Iscador Group 13 horses (41%) showed an improvement (at least 50% tumour remission). Of these cases a number of 9 patients showed complete cure (28%). In the Control Group only 3 cases (14%; all with complete cure) were classified as improved (p<0.05). Discussion & Conclusions: Iscador® P extract (viscum album pini) represents a safe and efficient therapy alternative in Equine Sarcoids, particularly in cases of multi-focal character. In tumours grown in problematic localizations a therapy attempt with viscum album extracts should be conducted to avoid the risk of damage the intact surrounding tissue, particularly in the eye area. Iscador® P shows significantly better outcomes (cure and improvement) compared to placebo control. Younger horses and horses with more than 2 tumours are responding with higher PO rates than older patients and horses with 1 to 2 ES. This fact could support the control of multiple sarcoids which can be excised only by immense surgery expense. Further investigations to understand factors of response and non-.response have to be conducted in the future. Additionally, due to the good tolerance by the patients and the acceptance of long-term protocol the practicability of Iscador ® P therapy can be assumed. Younger horses responded more often with a positive outcome (PO) than older (Fig. 2a) with PO rates of 57%, 40% and 13% for horses being less than 5 years, 5 to 9 years and more than 9 years, respectively compared to PO 25%, 10% and 0% in control. A significant better therapy course appeared in the Iscador group for horses with >2 ES (Fig. 2b): a number of 12 of 25 horses (48%) showed an improvement, 9 of these a complete remission (36%) including ES localized at the critical areas (head and around the eyes) compared to 6% in the control group. Iscador® P extract (viscum album pini) represents a safe and efficient therapy alternative in Equine Sarcoids, particularly in cases of multi-focal character. In tumours grown in problematic localizations a therapy attempt with viscum album extracts should be conducted to avoid the risk of damage the intact surrounding tissue, particularly in the eye area. Iscador® P shows significantly better outcomes (cure and improvement) compared to placebo control. Younger horses and horses with more than 2 tumours are responding with higher PO rates than older patients and horses with 1 to 2 ES. This fact could support the control of multiple sarcoids which can be excised only by immense surgery expense. Further investigations to understand factors of response and non-.response have to be conducted in the future. Additionally, due to the good tolerance by the patients and the acceptance of long-term protocol the practicability of Iscador ® P therapy can be assumed

Exercise‐induced airflow changes in horses with asthma measured by electrical impedance tomography

Background: Equine asthma (EA) causes airflow impairment, which increases in severity with exercise. Electrical impedance tomography (EIT) is an imaging technique that can detect airflow changes in standing healthy horses during a histamine provocation test. Objectives: To explore EIT-calculated flow variables before and after exercise in healthy horses and horses with mild-to-moderate (MEA) and severe equine asthma (SEA). Animals: Nine healthy horses 9 horses diagnosed with MEA and 5 with SEA were prospectively included. Methods: Recordings were performed before and after 15 minutes of lunging. Absolute values from global and regional peak inspiratory (PIF, positive value) and expiratory (PEF, negative value) flows were calculated. Data were analyzed using a mixed model analysis followed by Bonferroni's multiple comparisons test to evaluate the impact of exercise and diagnosis on flow indices. Results: Control horses after exercise had significantly lower global PEF and PIF compared to horses with SEA (mean difference [95% confidence interval, CI]: 0.0859 arbitrary units [AU; 0.0339-0.1379], P < .001 and 0.0726 AU [0.0264-0.1188], P = .001, respectively) and horses with MEA (0.0561 AU [0.0129-0.0994], P = .007 and 0.0587 AU [0.0202-0.0973], P = .002, respectively). No other significant differences were detected. Conclusions and clinical importance: Electrical impedance tomography derived PIF and PEF differed significantly between healthy horses and horses with SEA or MEA after exercise, but not before exercise. Differences between MEA and SEA were not observed, but the study population was small

Prevalence and WGS-based characteristics of Staphylococcus aureus in the nasal mucosa and pastern of horses with equine pastern dermatitis.

BACKGROUND Many contributing factors are involved in the development of equine pastern dermatitis (EPD). Among the most frequently suspected is Staphylococcus aureus, known for its pathogenic potential in skin and soft tissue infections. We therefore investigated the association between S. aureus carriage and EPD. RESULTS One hundred five EPD-affected horses and 95 unaffected controls were examined for the presence of methicillin-resistant and -susceptible Staphylococcus aureus (MRSA and MSSA) on the pastern skin and in the nostrils. S. aureus isolates were cultivated from swab samples on selective MSSA and MRSA chromogenic agar and identified using MALDI-TOF MS. Isolates were analysed by Illumina whole genome sequencing for genetic relatedness (cgMLST, spa typing), and for the presence of antimicrobial resistance and virulence determinants. A markedly higher proportion of samples from EPD-affected horses proved positive for S. aureus, both from the pastern (59.0 % vs. 6.3 % in unaffected horses; P<0.001), and from the nose (59.0 % vs. 8.4 %; P<0.001). Isolates belonged to 20 sequence types (ST) with lineages ST15-t084 (spa) (18 %), ST1-t127 (13 %), and ST1-t1508 (12 %) being predominant. Eight S. aureus were MRSA ST398-t011 and ST6239-t1456, and contained the staphylococcal cassette chromosome SCCmecIVa. Antimicrobial resistance genes were almost equally frequent in pastern and in nasal samples, whereas some virulence factors such as the beta-hemolysin, ESAT-6 secretion system, and some enterotoxins were more abundant in isolates from pastern samples, possibly enhancing their pathogenic potential. CONCLUSIONS The markedly higher prevalence of S. aureus containing specific virulence factors in affected skin suggests their contribution in the development and course of EPD

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Quantitative gait analysis before and after a cross-country test in a population of elite eventing horses.

Early recognition of lameness is crucial for injury prevention. Quantitative gait analysis systems can detect low-grade asymmetries better than the human eye and may be useful in early lameness recognition. The aims of this study were 1) to investigate the frequency of gait asymmetries based on head and pelvic movement in elite eventing horses using inertial mounted measurement units and 2) to assess the association between asymmetries and muscle enzymes and blood lactate (LA) levels post exercise. Movement asymmetry of the head, wither, and pelvis were quantified in 33 elite eventing horses prior to and one day after the cross-country test of three Concours Complet International (CCI3* and CCI4*) events held three weeks apart. The effects of LA concentration immediately after completion of the cross-country course and of serum levels of creatine kinase (CK) and aspartate amino-transferase (AST) four hours post-exercise on gait asymmetry parameters were tested with linear models. A total of 58% and 77% of the 33 horses exhibited gait asymmetries that exceeded published threshold values before and after the cross-country course, respectively. The magnitude of pre-existing gait asymmetries was not significantly increased after the cross-country test and no associations with post-exercise levels of CK, AST, or LA were detected. The stride duration was significantly shorter the day following the cross-country test and was associated with LA, the age and the weight of the horses. In conclusion, a majority of the horses studied presented gait asymmetries and strenuous exercise resulted in decreased stride duration but did not worsen gait asymmetries

Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells

The transcriptomic profile of a cell population can now be studied at the cellular level using single-cell mRNA sequencing (scRNA-seq). This novel technique provides the unprecedented opportunity to explore the cellular composition of the bronchoalveolar lavage fluid (BALF) of the horse, a species for which cell type markers are poorly described. Here, scRNA-seq technology was applied to cryopreserved equine BALF cells. Analysis of 4,631 cells isolated from three asthmatic horses in remission identified 16 cell clusters belonging to six major cell types: monocytes/macrophages, T cells, B/plasma cells, dendritic cells, neutrophils and mast cells. Higher resolution analysis of the constituents of the major immune cell populations allowed deep annotation of monocytes/macrophages, T cells and B/plasma cells. A significantly higher lymphocyte/macrophage ratio was detected with scRNA-seq compared to conventional cytological differential cell count. For the first time in horses, we detected a transcriptomic signature consistent with monocyte-lymphocyte complexes. Our findings indicate that scRNA-seq technology is applicable to cryopreserved equine BALF cells, allowing the identification of its major (cytologically differentiated) populations as well as previously unexplored T cell and macrophage subpopulations. Single-cell gene expression analysis has the potential to facilitate understanding of the immunological mechanisms at play in respiratory disorders of the horse, such as equine asthma