Application of locking compression plates as type 1 external fixators to treat unilateral mandibular fractures in four equids and one dromedary

Objectives To describe a novel fixation of open, unstable, unilateral mandibular fractures applying a locking compression plate as an external skeletal fixator (ESF-LCP). Animals Four horses and one dromedary. Study design Short case series. Methods Animals presented with unstable, open, unilateral fractures of the mandible. Fracture fixation was performed under general anesthesia. A 4.5/5.5 narrow LCP was applied externally above the level of the skin and combined with intraoral tension band wiring. Results Fracture fixation was achieved successfully using an ESF-LCP. Minimal tissue manipulation was required during application and removal of the construct. The ESF-LCPs provided adequate access to the wounds at the fracture site, were well tolerated, and did not interfere with any objects in the animals’ environment. Mild drainage at the screw-skin interface developed in all cases, requiring early implant removal due to surgical site infection in one case. The use of longer plates was associated with superficial pressure necrosis of the skin in the masseter area in two cases. Implants were removed after 3 to 12 weeks, and the long-term functional outcome after 11 to 41 (median 13) months was good in all cases. Conclusion Stabilization of mandibular fractures with ESF-LCP led to good outcomes in this case series. The use of longer plates positioned more caudally and in a ventrolateral position seemed associated with surgical site infection and pressure necrosis of the skin. Clinical significance Use of a locking compression plate as an external skeletal fixator seems to offer a viable alternative to treat unilateral mandibular fractures, especially when these are open and/or infected

Computed tomographic imaging and surgical management of distal insertional avulsion fragments of the caudal cruciate ligament in four horses

Objective To describe cases with caudal cruciate ligament (CdCL) avulsion fragments diagnosed based on computed tomography (CT) examination and report on arthroscopic fragment removal. Animals Four Warmblood horses with hindlimb lameness and osseous fragments located in the caudal medial femorotibial joint (mFTJ). Study design Short case series. Methods CT and arthroscopic evaluation of the caudal mFTJ were performed. The caudal mFTJ and the insertion of the CdCL on the tibia were assessed and removal of the avulsion fragments was attempted in three horses using a cranial intercondylar approach. Results The fragment was not accessible via caudomedial approaches in one horse. A cranial intercondylar approach was used in three horses, allowing removal of the intra-articular fragment in two horses, and removal of two-thirds of the proximal fragment in the last horse. Acute, profuse, arterial bleeding occurred in this horse during surgery with transient postoperative soft tissue swelling. Comorbidities included medial femoral condyle cartilage defects (3), cranial cruciate ligament lesions (2), and medial collateral ligament lesions (2). Horses were followed up for 16 months (median, range 11–28 months), at which point all were back in ridden exercise; owners' satisfaction was good. Conclusion CT examination confirmed the diagnosis and allowed evaluation of the stifle joint for comorbidities. A cranial intercondylar arthroscopic approach facilitated the removal of CdCL insertional avulsion fragments, although not always complete. Clinical significance A cranial intercondylar approach can allow access to CdCL avulsion fragments, but complications and incomplete removal remain possible

Objective evaluation for analgesia of the distal interphalangeal joint, the navicular bursa and perineural analgesia in horses with naturally occurring forelimb lameness localised to the foot

Background The outcome and interpretation of intra-synovial diagnostic analgesia of the distal interphalangeal joint (DIPJ) and the navicular bursa (NB) remain in dispute, and no objective studies have been carried out to establish the percentage of improvement over time from these two analgesia techniques. Objectives To investigate the qualitative and time-dependent outcome of DIPJ-A and NB-A in naturally occurring forelimb lameness. Study design Case series. Methods Twenty-three clinical cases with forelimb lameness were evaluated objectively using a body mounted inertial sensor system (BMIS). Lameness was localised to the foot with a palmar digital nerve block and/or an abaxial sesamoidean nerve block on day 1, and analgesia of the DIPJ (DIPJ-A) and NB (NB-A) were performed on days 2 and 3. Improvement following perineural analgesia was measured after 10 min and intra-synovial blocks after 2-, 5- and 10-min. Horses with at least 70% improvement measured objectively after diagnostic analgesia were included in the study. Results There was no significant association between improvement following perineural analgesia and the DIPJ-A and NB-A. The mean improvement in the lameness differed between DIPJ-A and NB-A at 2 min (p < 0.001) and at 5 min (p = 0.04), and it was no longer observed after 10 min (p = 0.06). A positive NB-A produced a high degree of improvement that remained stable, whereas the DIPJ-A improved over time. Main limitations Perineural and intra-synovial analgesia were performed without contrast medium to assess the diffusion of mepivacaine. Conclusions Our results suggest that perineural analgesia is not reliable enough to differentiate pain originating from DIPJ and NB. Early evaluation of the DIPJ-A and NB-A can determine the origin of the pain. An improvement following NB-A was constant over time, but an improvement following DIPJ-A varied by up to 10 min

Diagnosis and outcome following tenoscopic surgery of the digital flexor tendon sheath in German sports and pleasure horses

Background Digital flexor tendon sheath (DFTS) pathology is an important cause of lameness in horses. The outcome after surgical treatment is variable and depends on the exact diagnosis. Objectives To (1) describe the prevalence of lesions associated with lameness caused by nonseptic DFTS tenosynovitis in a large population of German sports and pleasure horses; (2) determine the sensitivity and specificity of diagnostic imaging techniques for identifying lesions within the DFTS with tenoscopic diagnosis being the gold standard; (3) explore associations between tenoscopically diagnosed lesions and signalment, purpose, and limb affected; and (4) describe the outcome following DFTS tenoscopy with nonseptic DFTS tenosynovitis in this population. Study design Retrospective case series. Methods Medical records of horses admitted for tenoscopic surgery of nonseptic DFTS tenosynovitis between 2011 and 2020 were reviewed. Follow‐up information was obtained via telephone contact. Sensitivity and specificity of ultrasonography and contrast tenography were determined using tenoscopic diagnosis as gold standard and univariable analysis was used to explore associations between signalment, case history, and tenoscopic diagnosis. Results Medical records from 131 horses were retrieved, of which 8 horses had bilateral disease and 6 horses were presented for tenoscopy on two separate occasions (3 for tenoscopy in the same limb, 3 in a different limb), thus, making a total of 145 limbs. Lesions were most commonly diagnosed in the deep (DDFT; n = 55 limbs) and superficial (SDFT; n = 55 limbs) digital flexor tendons. Manica flexoria (MF) lesions were detected in 44 limbs and palmar/plantar annular ligament (PAL) constriction in 99 limbs. In 36 limbs, only one structure within the DFTS was injured, whereas in 109 limbs a combination of lesions was noted, the most common being the combination of a SDFT lesion with PAL constriction. All affected limbs were examined with diagnostic ultrasonography; contrast tenography was performed in 86 limbs. For diagnosis of MF and DDFT tears, tenography was more sensitive (89% [confidence interval, CI: 65.4%–95.2%]; 72% [CI: 46.4%–89.3%], respectively) than specific (64% [CI: 52.5%–77.6%]; 53% [CI: 42.2%–73.3%], respectively) whereas ultrasonography was more specific (92% [CI: 84.5%–96.3%]; 92% [CI: 83.6%–96.0%]) with lower sensitivity (64% [CI: 47.7%–77.2%]; 54% [CI: 39.5%–67.9%]). For SDFT lesions, ultrasonography was highly specific (94% [CI: 86.9%–97.9%]) with lower sensitivity (66% [CI: 51.3%–77.4%]). Follow‐up information following first surgery was obtained for 118 horses (132 limbs): 18 (15.3%) of 118 horses remained chronically lame, 40 (33.8%) performed at a reduced level and 60 (50.8%) performed at the same or higher level following rehabilitation after tenoscopy. Horses with DDFT lesions had the poorest outcomes with only 36.6% returning to the same or higher level of exercise. Main limitations Retrospective analysis of clinical records and subjective outcome assessment based on owner follow‐up with potential recall bias. Findings on diagnostic imaging are impacted by many factors including equipment quality and operator expertise and experience. Conclusion Diagnostic imaging techniques were complimentary and contrast tenography was sensitive and ultrasonography was specific for the diagnosis of MF and DDFT lesions. Following tenoscopic surgery for nonseptic tenosynovitis of the DFTS, approximately half the cases were able to return to preinjury level of exercise

Local and systemic effect of transfection-reagent formulated DNA vectors on equine melanoma

Background Equine melanoma has a high incidence in grey horses. Xenogenic DNA vaccination may represent a promising therapeutic approach against equine melanoma as it successfully induced an immunological response in other species suffering from melanoma and in healthy horses. In a clinical study, twenty- seven, grey, melanoma-bearing, horses were assigned to three groups (n = 9) and vaccinated on days 1, 22, and 78 with DNA vectors encoding for equine (eq) IL-12 and IL-18 alone or in combination with either human glycoprotein (hgp) 100 or human tyrosinase (htyr). Horses were vaccinated intramuscularly, and one selected melanoma was locally treated by intradermal peritumoral injection. Prior to each injection and on day 120, the sizes of up to nine melanoma lesions per horse were measured by caliper and ultrasound. Specific serum antibodies against hgp100 and htyr were measured using cell based flow- cytometric assays. An Analysis of Variance (ANOVA) for repeated measurements was performed to identify statistically significant influences on the relative tumor volume. For post-hoc testing a Tukey-Kramer Multiple-Comparison Test was performed to compare the relative volumes on the different examination days. An ANOVA for repeated measurements was performed to analyse changes in body temperature over time. A one-way ANOVA was used to evaluate differences in body temperature between the groups. A p–value < 0.05 was considered significant for all statistical tests applied. Results In all groups, the relative tumor volume decreased significantly to 79.1 ± 26.91% by day 120 (p < 0.0001, Tukey-Kramer Multiple-Comparison Test). Affiliation to treatment group, local treatment and examination modality had no significant influence on the results (ANOVA for repeated measurements). Neither a cellular nor a humoral immune response directed against htyr or hgp100 was detected. Horses had an increased body temperature on the day after vaccination. Conclusions This is the first clinical report on a systemic effect against equine melanoma following treatment with DNA vectors encoding eqIL12 and eqIL18 and formulated with a transfection reagent. Addition of DNA vectors encoding hgp100 respectively htyr did not potentiate this effect

Diagnostic endoscopy of the navicular bursa using a needle endoscope by direct or transthecal approach: A comparative cadaver study

OBJECTIVES: To compare the practicability, visualization of structures, and iatrogenic damage of direct and transthecal approaches to the navicular bursa for diagnostic needle endoscopy. STUDY DESIGN: Descriptive study. SAMPLE POPULATION: Equine cadaver forelimbs (n = 30). METHODS: Direct and transthecal approaches for insertion of a needle endoscope into the navicular bursa were performed. Video recordings of endoscopic procedures were assessed to determine all structures visualized within the navicular bursa. Number of attempts to gain access to the navicular bursa and total time for insertion and examination were recorded. Distribution and severity of iatrogenic lesions were assessed and scored after dissection. RESULTS: There were no statistical differences for number of attempts or time needed for insertion and examination between direct and transthecal approaches. The direct approach offered significantly increased visibility of the ipsilateral abaxial and proximal margins of the navicular bone, and ipsilateral collateral sesamoidean ligament. Iatrogenic lesions were superficial and focal, regardless of approach taken, or whether a blunt or sharp trocar tip was used. CONCLUSIONS: The direct approach provided significantly better visualization of the ipsilateral structures within the navicular bursa compared to the transthecal approach. Needle endoscopy offers a reliable technique to evaluate the navicular bursa and may complement or replace other diagnostic modalities in horses with lameness localized to the navicular region

Increased FOXP3 expression in tumour-associated tissues of horses affected with equine sarcoid disease.

Recent studies suggest that regulatory T cells (Tregs) are associated with disease severity and progression in papilloma virus induced neoplasia. Bovine papilloma virus (BPV) is recognised as the most important aetiological factor in equine sarcoid (ES) disease. The aim of this study was to compare expression levels of Treg markers and associated cytokines in tissue samples of ES-affected equids with skin samples of healthy control horses. Eleven ES-affected, and 12 healthy horses were included in the study. Expression levels of forkhead box protein 3 (FOXP3), interleukin 10 (IL10), interleukin 4 (IL4) and interferon gamma (IFNG) mRNA in lesional and tumour-distant samples from ES-affected horses, as well as in dermal samples of healthy control horses were measured using quantitative reverse transcription polymerase chain reaction (PCR). Expression levels were compared between lesional and tumour-distant as well as between tumour-distant and control samples. Furthermore, BPV-1 E5 DNA in samples of ES-affected horses was quantified using quantitative PCR, and possible associations of viral load, disease severity and gene expression levels were evaluated. Expression levels of FOXP3, IL10 and IFNG mRNA and BPV-1 E5 copy numbers were significantly increased in lesional compared to tumour-distant samples. There was no difference in FOXP3 and cytokine expression in tumour-distant samples from ES- compared with control horses. In tumour-distant samples viral load was positively correlated with IL10 expression and severity score. The increased expression of Treg markers in tumour-associated tissues of ES-affected equids indicates a local, Treg-induced immune suppression

Comprehensive characteristics of microRNA expression profile of equine sarcoids

Equine sarcoids are the most common neoplasms occurring in horses. Despite frequent occurrence, they are still not well described at the molecular level. Thus, in the present study, we performed a comprehensive comparative analysis of sarcoid miRNAome profile to identify aberrantly expressed microRNAs, along with their structural variants, potentially useful as biomarkers and, in a wider perspective, broaden the knowledge about this tumor and underlying mechanisms. To this end, we conducted next generation sequencing and as a result we identified both known and potentially novel miRNAs. Differential expression analysis revealed the existence of almost one hundred miRNAs being over- or underexpressed in sarcoids in comparison to healthy tissue (p-adj<0.05), of which many are known for their involvement in processes crucial for neoplastic transformation. Among upregulated miRNAs there were those associated with decreased cell adhesion abilities as well as engaged in global protein production, while downregulation of some miRs i.a. increased cell expansion abilities. Moreover, we identified altered expression levels of miRNA variants (isomiRs) between the investigated tissues. Further analysis revealed that 5' isomiRs comprise different seed sequences leading to target gene switching followed by activation of different biological pathways. Our results are the first which revealed the complexity of microRNA profiles in equine sarcoids and skin tissue, along with the dynamism of their growing in importance concomitants, namely isomiRs. They also showed miRNA molecules and biological pathways important from the sarcoid oncogenesis point of view

DNA hypomethylation and oxidative stress-mediated increase in genomic instability in equine sarcoid-derived fibroblasts

It is widely accepted that equine sarcoid disease, the most common skin associated neoplasm in equids, is induced by bovine papillomavirus (BPV-1). Although BPV-1 DNA has been found in almost all examined sarcoids so far, its detailed impact on the horse's host cell metabolism is largely unknown. We used equine fibroblast cell lines originating from sarcoid biopsies to study BPV-1-associated changes on DNA methylation status and oxidative stress parameters. Sarcoid-derived fibroblasts manifested increased proliferation in vitro, transcriptional rDNA activity (NORs expression) and DNA hypomethylation compared to control cells. Cells isolated from equine sarcoids suffered from oxidative stress: the expression of antioxidant enzymes was decreased and the superoxide production was increased. Moreover, increased ploidy, oxidative DNA damage and micronuclei formation was monitored in sarcoid cells. We postulate that both altered DNA methylation status and redox milieu may affect genomic stability in BPV-1-infected cells and in turn contribute to sarcoid pathology

Genomic landscape of copy number variation and copy neutral loss of heterozygosity events in equine sarcoids reveals increased instability of the sarcoid genome.

Although they are the most common neoplasms in equids, sarcoids are not fully characterized at the molecular level. Therefore, the objective of this study was to characterize the landscape of structural rearrangements, such as copy number variation (CNV) and copy neutral loss of heterozygosity (cnLOH), in the genomes of sarcoid tumor cells. This information will not only broaden our understanding of the characteristics of this genome but will also improve the general knowledge of this tumor and the mechanisms involved in its generation. To this end, Equine SNP64K Illumina microarrays were applied along with bioinformatics tools dedicated for signal intensity analysis. The analysis revealed increased instability of the genome of sarcoid cells compared with unaltered skin tissue samples, which was manifested by the prevalence of CNV and cnLOH events. Many of the identified CNVs overlapped with the other research results, but the simultaneously observed variability in the number and sizes of detected aberrations indicated a need for further studies and the development of more reliable bioinformatics algorithms. The functional analysis of genes co-localized with the identified aberrations revealed that these genes are engaged in vital cellular processes. In addition, a number of these genes directly contribute to neoplastic transformation. Furthermore, large numbers of cnLOH events identified in the sarcoids suggested that they may play no less significant roles than CNVs in the carcinogenesis of this tumor. Thus, our results indicate the importance of cnLOH and CNV in equine sarcoid oncogenesis and present a direction of future research