The assessment of paraspinal muscle epimuscular fat in participants with and without low back pain: A case-control study.

It remains unclear whether paraspinal muscle fatty infiltration in low back pain (LBP) is i) solely intramuscular, ii) is lying outside the epimysium between the muscle and fascial plane (epimuscular) or iii) or combination of both, as imaging studies often use different segmentation protocols that are not thoroughly described. Epimuscular fat possibly disturbs force generation of paraspinal muscles, but is seldomly explored. This project aimed to 1) compare epimuscular fat in participants with and without chronic LBP, and 2) determine whether epimuscular fat is different across lumbar spinal levels and associated with BMI, age, sex and LBP status, duration or intensity. Fat and water lumbosacral MRIs of 50 chronic LBP participants and 41 healthy controls were used. The presence and extent of epimuscular fat for the paraspinal muscle group (erector spinae and multifidus) was assessed using a qualitative score (0-5 scale; 0 = no epimuscular fat and 5 = epimuscular fat present along the entire muscle) and quantitative manual segmentation method. Chi-squared tests evaluated associations between qualitative epimuscular fat ratings and LBP status at each lumbar level. Bivariate and partial spearman's rho correlation assessed relationships between quantitative and qualitative epimuscular fat with participants' characteristics. Epimuscular fat was more frequent at the L4-L5 (X2 = 13.781, p = 0.017) and L5-S1 level (X2 = 27.825, p < 0.001) in participants with LBP compared to controls, which was not found for the higher lumbar levels. The total qualitative score (combined from all levels) showed a significant positive correlation with BMI, age, sex (female) and LBP status (r = 0.23-0.55; p < 0.05). Similarly, the total area of epimuscular fat (quantitative measure) was significantly correlated with BMI, age and LBP status (r = 0.26-0.57; p < 0.05). No correlations were found between epimuscular fat and LBP duration or intensity. Paraspinal muscle epimuscular fat is more common in chronic LBP patients. The functional implications of epimuscular fat should be further explored.

Radiological features of arterial channels in the equine third phalanx measured using a novel customized software represent changes of laminitis.

OBJECTIVE: To identify and measure radiolucencies at the solear margin of the distal phalanx in radiographs of healthy and laminitic hooves. SAMPLE: Clinical records and dorsoproximal-palmarodistal radiographs of equine forelimbs with radiological diagnoses of either laminitis (n = 40, L) or navicular syndrome (n = 40, NS). METHODS: Outlines of the radiolucent structures at the solar margin were drawn in ImageJ, and a customized novel plugin "Arteries Analyzer/ImageJ" was used for measurements. The diverging radiolucencies outside the terminal arc of the distal phalanx were differentiated as arterial channels (open at the solear margin) and ellipses (closed at the solear margin). Comparisons between L and NS, between distal phalanges with and without ellipses, and of arterial channels and ellipses in areas were compared using Wilcoxon and the Mann-Whitney U tests, respectively. The reliability and repeatability of the method were tested using Friedman's test. RESULTS: Fewer arterial channels but more ellipses were identified in L than in NS. In phalanges with ellipses (n = 47), the number of ellipses and the number of arterial channels were negatively correlated (PCC -0.181, P = .224). The number of ellipses correlated positively with the severity of laminitis (PCC 0.495, P < .001; n = 80) and with the degree of rotation of the distal phalanx (PCC 0.392, P < .001; n = 80). CLINICAL RELEVANCE: The software tool successfully measured arterial channels and ellipses outlined by the evaluators. Results indicate that healthy arteries develop into pathological ellipses in laminitic feet. This may be used to complement the interpretation of radiographs and support clinical decision-making.

Health Promotion for Students of Veterinary Medicine: A Preliminary Study on Active Microbreaks and Ergonomics Education.

Occupational hazards, such as psychosocial stressors, physical injuries from human-animal interactions, and physically demanding work tasks, are common in the veterinary profession, and musculoskeletal discomfort and pain (MDP) may already be present in veterinary undergraduates. This preliminary study investigates the effects of very short, active interventions, called microbreaks, in 36 veterinary students. At the start, participants had a high prevalence of MDP, especially in the neck and lower back. Within a 12-week observational period, 6 weeks of active intervention comprised teaching microbreaks (nine strengthening, stretching, and relaxation exercises; 30-90 s each) and a weekly veterinary-specific ergonomics education and discussion. After the intervention, participants reported fewer painful body regions and an increase in their self-efficacy in potentially painful, risky, or dangerous human-animal interactions. After the 12-week observational period, participants had increased self-efficacy in the maintenance of physical health and self-protection but decreased self-efficacy in healing injuries after veterinary human-animal interactions. Participants felt to have increased and decreased control over dangerous situations with dogs and horses, respectively, although self-efficacy in handling horses increased. Participants integrated microbreaks well into their undergraduate activities and rated the topic relevant to their (later) profession. This should encourage the inclusion of similar programs in undergraduate curricula.

Comparison of gluteus medius muscle activity in Haflinger and Noriker horses with polysaccharide storage myopathy.

Type 1 polysaccharide storage myopathy caused by genetic mutation in the glycogen synthase 1 gene is present in many breeds including the Noriker and Haflinger horses. In humans, EMG has already been used to document changes in the muscle activity patterns of patients affected by human glycogen storage disorders. Therefore, the aim of the present study was to describe gluteus muscle activity with surface electromyography (sEMG) in Haflinger and Noriker horses with known GYS1 mutation status during walk and trot. Thirty-two horses (11 Haflinger and 21 Noriker horses) with homozygous non-affected (GG), heterozygous affected (GA) and homozygous affected (AA) status of GYS1 mutation without overt clinical signs of any myopathy were selected for the current study. Using surface electromyography gluteus medius muscle activity at walk and at trot was measured, and muscle activity was described in relation to the maximum observed value at the same sensor and the same gait. In order to further describe the signals in detail comprising both frequencies and amplitudes, the crossings through the baseline and the 25, 50 and 75 percentile lines were determined. The result of the relative muscle activity did not show a consistent difference between affected and non-affected horses. Genetically affected (GA and AA) horses showed significantly less density of muscle activity for both gaits and horse breeds except for the crossings per second at the baseline and 75 percentile at walk in the Haflinger horses and 75 percentile at trot in the Noriker horses. The medians of all calculated density values were significantly lower in the GA Haflingers compared to the GG Haflingers (p = 0.012) and also in the AA Norikers compared to the GG Norikers (p = 0.011). Results indicate that the GYS1 mutation reduces the number of functional muscle fibres detected by sEMG measurements even in the absence of overt clinical signs.

Movement Caused by Electrical Stimulation of the Lumbosacral Region in Standing Horses.

Electrical stimulation is commonly used as a modality for physical therapy in human and veterinary medicine. However, studies measuring the movement generated by electrical stimulation in horses are rare. The present study therefore evaluates the range of movement provoked by a commercially available physical therapy unit (FES310) and contrasts it with the movement generated by manually induced pelvic inclination (back rounding). Ten horses were tested on three measurement days over one week. Electrical stimulation was applied via a back treatment pad (belonging to the FES310 system) containing six electrodes (three on either side of the spine) placed over the lumbosacral region. This system produced a pulsed, biphasic electrical stimulation in a rectangular waveform which was gradually increased to a maximum of 10 volts. Before and after electrical stimulation testing, manual pelvic inclination was achieved by pressure on two points lateral to the root of the tail. Muscle tone and lameness were evaluated before and after treatments. Skinfold thickness, body condition score, and body mass were measured to detect possible confounding factors. Using kinematics, the angle ranges during movement of ten three-dimensional angles of the trunk, the pelvis, and the hind limbs were further analyzed. Movement was produced with manual stimulation in every tested individual on all measurement days and with electrical stimulation on at least one measurement day. The electrical stimulation led to significantly (P < .05) smaller angle ranges which were 15 %-57 % of the median of the manually stimulated movement. Strong positive correlations between angle ranges of the electrically generated movement were found for the hind limbs implicating their involvement in the movement created. Correlations between skinfold thickness, body condition score, and body mass with the angle ranges were weak and not significant. Before and after electrical and manual stimulation, muscle tone and lameness were similar. In the present study, both electrical and manual stimulation were proven to produce significant trunk and hind limb movement. Within this study's electrical stimulation treatment protocol, the movement generated by electrical stimulation was significantly less than the movement caused by manual pelvic inclination. However, electrical stimulation could easily be applied over a longer period and in a higher frequency than it would be possible for manual pelvic inclination. This treatment shows potential for stabilization and or mobilization of the lumbosacral region, although its efficiency as a therapeutic tool and its effect on specific orthopedic problems and is to be evaluated in further research.

Equine Hoof Canker: Bovine Papillomavirus Infection Is Not Associated With Impaired Keratinocyte Differentiation.

Impaired keratinocyte differentiation has recently been suggested as a key event in equine hoof canker development. Koilocytotic appearance of keratinocytes, one of the most characteristic morphological alterations in hoof canker tissue, is also a common marker for papillomavirus (PV) infection, and bovine PV-1 and/or -2 (BPV-1/2) has previously been detected in equine canker patients. Therefore, the present study aimed to correlate the frequency and severity of koilocytotic keratinocytes with BPV detection in hoof canker samples. Hoof tissue of 5/18 canker-affected horses and 2/6 control horses tested positive for BPV-1/2 DNA using polymerase chain reaction. Thus, no association between the presence of BPV-1/2 papillomaviral DNA and koilocytotic appearance was found. Proteins associated with but not specific for PV infection were also investigated. Using immunohistochemistry, specific adhesion molecules (E-cadherin and ß-catenin) and intermediate filaments (keratins 6 and 14) important for intact epidermal barrier function and keratinocyte differentiation were documented in control samples (n = 6) and in hoof canker tissue samples (n = 19). Altered expression patterns of intermediate filaments and adhesion molecules were demonstrated in canker tissue, confirming the importance of incomplete keratinocyte differentiation, as well as the crucial role of keratinocyte differentiation in hoof canker.

Broad-Spectrum Cephalosporin-Resistant Klebsiella spp. Isolated from Diseased Horses in Austria.

The aim of the present study was to investigate the diversity of broad-spectrum cephalosporin-resistant Klebsiella spp. isolated from horses in Austria that originated from diseased horses. A total of seven non-repetitive cefotaxime-resistant Klebsiella sp. isolates were obtained during diagnostic activities from autumn 2012 to October 2019. Antimicrobial susceptibility testing was performed. The isolates were genotyped by whole-genome sequencing (WGS). Four out of seven Klebsiella isolates were identified as K. pneumoniae, two as K. michiganensis and one as K. oxytoca. All isolates displayed a multi-drug resistant phenotype. The detection of resistance genes reflected well the phenotypic resistance profiles of the respective isolates. All but one isolate displayed the extended-spectrum ß-lactamases (ESBL) phenotype and carried CTX-M cefotaximases, whereas one isolate displayed an ESBL and AmpC phenotype and carried cephamycinase (CMY)-2 and sulfhydryl variable (SHV)-type b and Temoniera (TEM) ß-lactamases. Among Klebsiella pneumoniae isolates, for different sequence types (ST) could be detected (ST147, ST307, ST1228, and a new ST4848). Besides resistance genes, a variety of virulence genes, including genes coding for yersiniabactin were detected. Considering the high proximity between horses and humans, our results undoubtedly identified a public health issue. This deserves to be also monitored in the years to come.

Endotoxin-induced changes in expression of cyclooxygenase isoforms in the lamellar tissue of extracorporeally haemoperfused equine limbs.

Angiogenesis and sepsis-related equine laminitis have several features in common. Both events can be induced by endotoxin (lipopolysaccharide- LPS) and both are associated with increased expression of the enzyme cyclooxygenase (COX), of which two isoforms (COX-1 and COX-2) exist. To examine the causal relationship between LPS exposure and COX expression and to investigate the tissue distribution of COX in the LPS-exposed tissue, the technique of extracorporeal haemoperfusion of isolated equine forelimbs was utilized. Perfusion was performed for 10 hr under physiological conditions (control-perfused limbs, n = 5) and with addition of 80 ng/L of endotoxin (LPS-perfused limbs; n = 5). After perfusion, samples of lamellar tissue were collected from the dorsal aspect of the hoof wall. Additional control samples were collected from three non-perfused limbs. Immunohistochemical analysis was performed using antibodies against COX-1 and COX-2, and intensity of immunohistochemical staining was scored for each isoform. In the lamellar tissue of control- and LPS-perfused limbs, there was no significant difference in COX-1 staining intensity and distribution, whereas COX-2 expression was significantly increased in LPS-perfused limbs (especially in endothelial cells, fibroblasts and intravasal leucocytes as well as in epidermal basal cells at the base of the primary epidermal lamellae). These results suggest that COX-2 and its metabolites are involved in the initiation of pathological changes seen in sepsis-associated events such as sepsis-related laminitis. In such cases, COX-2 could therefore be an important therapeutic target; however, early therapy may be required as increase in COX-2 expression occurs within 10 hr after LPS exposure.

Use of Physical Self-Experience for Teaching Lameness Evaluation: Short-Term Effects on Lameness Evaluation of Horses with Mild Forelimb Lameness by Novice Veterinary Students.

Lameness evaluation (LE) is an important veterinary skill, but it can be a challenging task to teach. Acoustic and visual input in conjunction with acting are the three major learning channels for acquiring new information. Acting (e.g., physical self-experience) has not been reported as a didactic tool for LE; therefore, we investigated two physical self-experience techniques, imitation of lameness during LE instruction and head nod during LE. Undergraduate veterinary students (N = 78) who had not yet received any instructions for LE underwent standard instruction for LE, followed by instructions to head nod, instructions to imitate lameness, or neither. Students' subjective LE skills were then tested against those of senior clinicians on two horses that were either sound or mildly forelimb lame. In more than 80%, there was agreement on the presence or absence of lameness independent of instruction, and we found no significant evaluation differences between didactic strategies. Specific instructions for head nodding did not significantly influence the quality of LE, possibly because head nodding was widespread even when no specific instructions had been given. Veterinary students who had been taught to imitate lameness without instructions to head nod were consistently closest to the assessment of the senior clinicians, and in walk these students were significantly better at LE than students additionally instructed to head nod. This finding indicates that imitation of lameness, but not necessarily head nodding, may be beneficial for understanding body movement such as mild equine forelimb lameness.

Trot Accelerations of Equine Front and Hind Hooves Shod with Polyurethane Composite Shoes and Steel Shoes on Asphalt.

The present study investigated accelerations of the front and hind hooves of horses comparing two different shoe types. A standard steel shoe, with studs, pins, and in some instances with toe grabs, was compared to a steel shoe covered on the bottom with a layer of polyurethane. Four horses were used; they trotted in hand on an asphalt track at their self-selected speed. The results showed significantly reduced decelerations during the stance phase with the polyurethane-covered shoes (10th percentile median steel -2.77 g, polyurethane -2.46 g; p = 0.06) and significantly increased decelerations in front hooves compared to hind hooves with steel shoes (70th percentile median -1.04 g front hooves, 0.12 g hind hooves, p = 0.04). Horses trotted faster using longer strides with the polyurethane-covered shoes compared to the steel shoes. The results show that effects of shoe types should be investigated simultaneously in front and hind hooves, and that PU shoes may aid in reducing the overload present in the front limbs of horses.

Establishment of a model for equine small intestinal disease: effects of extracorporeal blood perfusion of equine ileum on metabolic variables and histological morphology - an experimental ex vivo study.

BACKGROUND: In horses a number of small intestinal diseases is potentially life threatening. Among them are Equine Grass Sickness (EGS), which is characterised by enteric neurodegeneration of unknown aetiology, as well as reperfusion injury of ischaemic intestine (I/R), and post-operative ileus (POI), common after colic surgery. The perfusion of isolated organs is successfully used to minimize animal testing for the study of pathophysiology in other scenarios. However, extracorporeal perfusion of equine ileum sourced from horses slaughtered for meat production has not yet been described. Therefore the present study evaluated the potential of such a model for the investigation of small intestinal diseases in an ex vivo and cost-efficient system avoiding experiments in live animals. RESULT: Nine ileum specimens were sourced from horses aged 1-10 years after routine slaughter at a commercial abattoir. Ileum perfusion with oxygenated autologous blood and plasma was successfully performed for 4 h in a warm isotonic bath (37.0-37.5 °C). Ileum specimens had good motility and overall pink to red mucosa throughout the experiment; blood parameters indicated good tissue vitality: 82 ± 34 mmHg mean arterial partial pressure of oxygen (pO2) compared to 50 ± 17 mmHg mean venous pO2, 48 ± 10 mmHg mean arterial partial pressure of carbon dioxide (pCO2) compared to 66 ± 7 mmHg venous pCO2 and 9.8 ± 2.8 mmol/L mean arterial lactate compared to 11.6 ± 2.7 mmol/L venous lactate. There was a mild increase in ileum mass reaching 105 ± 7.5% of the pre-perfusion mass after 4 hours. Histology of haematoxylin and eosin stained biopsy samples taken at the end of perfusion showed on average 99% (±1%) histologically normal neurons in the submucosal plexus and 76.1% (±23.9%) histologically normal neurons in the myenteric plexus and were not significantly different to control biopsies. CONCLUSION: Extracorporeal, normothermic perfusion of equine ileum over 4 h using autologous oxygenated blood/plasma perfusate showed potential as experimental model to test whether haematogenous or intestinal exposure to neurotoxins suspected in the pathogenesis of EGS can induce neuronal damage typical for EGS. Also, this model may allow investigations into the effect of pharmaceuticals on I/R injury, as well as into the pathogenesis of equine POI.

Increased genetic diversity of methicillin-resistant Staphylococcus aureus (MRSA) isolated from companion animals.

The aim of the present study was to investigate the diversity of methicillin-resistant Staphylococcus aureus (MRSA) that originated from Austrian companion animals during the last five-year period. A total of 90 non-repetitive MRSA isolates were obtained during diagnostic activities from autumn 2013 to autumn 2018. They originated from horses (n = 62), cats (n = 13), dogs (n = 10), rabbits (n = 2), a domestic canary, a zoo-kept hammer-headed bat (Hypsignathus monstrosus) and a semi-captive northern bald ibis (Geronticus eremita). Antimicrobial susceptibility testing was performed. All isolates were mecA-positive and mecC-negative. The isolates were genotyped by SCCmec, spa and dru typing, Multiple-Locus Variable number of tandem repeat Analyses (MLVA), S. aureus DNA microarray, and whole-genome sequencing (WGS). Eight sequence types (STs - ST398, ST5275 (new ST), ST225, ST8, ST22, ST152, ST1, and ST45), three SCCmec types (II, IV, and V), sixteen spa types (t003, t008, t011, t015, t032, t034, t1381, t1928, t1985, t223, t334, t355, t430, t6447, t6867, and t7105), fourteen dru types (dt10a, dt10az, dt10q, dt10r, dt11a, dt5e, dt6j, dt9a, dt9ak, dt9g, and four new types dt8as, dt7ak, dt4j, dt14n), and thirty-five MLVA types were detected. WGS-based core genome MLST (cgMLST) displayed five main clusters. Compared to the time period 2004-2013, the results of the present study show not only a higher diversity among the MRSA isolates within the population of Austrian companion animals, but also the introduction of new clones. Although ST398 isolates remained predominant, mainly due to high presence of this lineage among horses, increasing isolation rates of human-associated MRSA clones were observed in cats and dogs.

Endotoxin-induced changes of type VII collagen- cleaving matrix metalloproteinases in lamellar tissue of extracorporeally perfused equine limbs.

OBJECTIVE To investigate the effect of lipopolysaccharide (LPS) on type VII collagen- cleaving matrix metalloproteinases (MMPs) in the lamellar tissue of extracorporeally perfused equine limbs. SAMPLE 10 right forelimbs and 3 left forelimbs collected from 10 adult horses after slaughter at a licensed abattoir. PROCEDURES Extracorporeal perfusion of the isolated equine limbs was performed for 10 hours under physiologic conditions (control-perfused limbs; n = 5) and with the addition of 80 ng of LPS/L of perfusate (LPS-perfused limbs; 5). Lamellar tissue specimens were then collected from the dorsal aspect of the hooves. Additionally, corresponding control specimens were collected from the 3 nonperfused left forelimbs. Immunohistochemical analysis was performed on paraffin-embedded tissue blocks with antibodies against total (latent and active) MMP-1, MMP-2, MMP-8, and MMP-9 as well as antibody against active MMP-9. Intensity of immunohistochemical staining was scored, and stain distribution in the lamellar tissue was noted. RESULTS Staining intensity of total and active MMP-9 was significantly increased in LPS-perfused versus control-perfused limbs. No such difference was identified for MMP-1, MMP-2, and MMP-8. CONCLUSIONS AND CLINICAL RELEVANCE Of the 4 MMPs that are capable of degrading type VII collagen, MMP-9 was the only one for which production increased in the lamellar tissue of isolated equine limbs perfused with versus without a clinically relevant concentration of LPS. These results suggested that MMP-9 may be involved in initiation of pathological changes in lamellar tissue in endotoxin-induced laminitis, whereas MMP-1, MMP-2, and MMP-8 may be less relevant.

Long term consistency and location specificity of equine gluteus medius muscle activity during locomotion on the treadmill.

BACKGROUND: The equine m. gluteus medius (GM) is the largest muscle of the horse, its main movement function is the extension of the hip joint. The objective of the present study was to measure equine GM activity in three adjacent locations on GM during walk and trot on a treadmill, in order to document potential differences. Fourteen Haflinger mares were measured using surface electromyography and kinematic markers to identify the motion cycles on three occasions over 16 weeks. The electrodes were placed on left and right gluteus medius muscle over the middle of its widest part and 5 cm lateral and medial of it. For data processing, electrical activity was normalised to its maximum value and timing was normalised to the motion cycle. A Gaussian distribution approach was used to determine up to 10 modes of focussed activity, and results were analysed separately for stance and swing phase of the ipsilateral hindlimb. RESULTS: Fair reliability was found for mean mode values (Cronbach's alpha = 0.66) and good reliability was found for mean mode locations (Cronbach's alpha = 0.71) over the three data collection days. The magnitude of muscle activity identified as mean mode value was much larger at trot than at walk, and mean mode value was significantly different between stance phases of walk and trot for all electrode positions (p < 0.01). The pattern of muscle activity identified as mean mode location was significantly different for walk and trot at all electrode positions, both during stance and swing phases (p < 0.001). This indicates the different timing pattern between the gaits. Results of the three electrode positions on the same muscle during each gait were not significantly different when comparing the same measurement. CONCLUSIONS: The middle of the equine GM does not show any indication of functional differentiation during walk and trot on a treadmill; this might be due to lack of segmentation as such, or due to lack of need for segmented use for these very basic main tasks of the muscle. The reliability of the sEMG measurements over several weeks was fair to good, an indication for the robustness of the methodology.

Assessment of tissue-specific cortisol activity with regard to degeneration of the suspensory ligaments in horses with pituitary pars intermedia dysfunction.

OBJECTIVE To identify signs of tissue-specific cortisol activity in samples of suspensory ligament (SL) and neck skin tissue from horses with and without pituitary pars intermedia dysfunction (PPID). SAMPLE Suspensory ligament and neck skin tissue samples obtained from 26 euthanized horses with and without PPID. PROCEDURES Tissue samples were collected from 12 horses with and 14 horses without PPID (controls). Two control horses had received treatment with dexamethasone; data from those horses were not used in statistical analyses. The other 12 control horses were classified as old horses (≥ 14 years old) and young horses (≤ 9 years old). Standard histologic staining, staining for proteoglycan accumulation, and immunostaining of SL and neck skin tissue sections for glucocorticoid receptors, insulin, 11ß hydroxysteroid dehydrogenase type 1, and 11ß hydroxysteroid dehydrogenase type 2 were performed. Findings for horses with PPID were compared with findings for young and old horses without PPID. RESULTS Compared with findings for old and young control horses, there were significantly more cells stained for glucocorticoid receptors in SL samples and for 11 ß hydroxysteroid dehydrogenase type 1 in SL and skin tissue samples from horses with PPID. Insulin could not be detected in any of the SL or skin tissue samples. Horses with PPID had evidence of SL degeneration with significantly increased proteoglycan accumulation. Neck skin tissue was found to be significantly thinner in PPID-affected horses than in young control horses. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that tissue-specific dysregulation of cortisol metabolism may contribute to the SL degeneration associated with PPID in horses.

Extracorporeal perfusion of isolated organs of large animals - Bridging the gap between in vitro and in vivo studies.

Since the early 20th century, extracorporeal perfusion of large animal organs has been used to investigate a broad variety of research questions, thereby overcoming common drawbacks of in vitro studies without suffering from ethical concerns associated with live animal research. The technique is in accordance with the 3R principles and represents an excellent opportunity to investigate the physiology of organs in detail under standardized conditions. It is also suitable for the translation of basic pre-clinical research into a more relevant arena prior to or avoiding live animal research altogether. Furthermore, organ perfusion has also been an important tool in developing methods of organ preservation for transplantation surgery. Yet, due to the nature of the experiments, only short-term observations can be made and while cells are still exposed to their regional secretome, the whole organ itself is isolated from the body and correlations between organ systems cannot be taken into consideration. This review gives an overview over the history of extracorporeal perfusion of large animal organs and limbs, highlighting major achievements in the field and discussing different experimental set-ups. Advantages and limitations of the technique are presented. Prospective future research perspectives, which may include tracking of specific cell types and study of their distinct behavior towards different stimuli, are given to illustrate the relevance of this method.

Initial adhesion of methicillin-sensitive and methicillin-resistant Staphylococcus aureus strains to untreated and electropolished surgical steel drill bits.

Electropolishing of stainless steel has been thoroughly investigated as a prophylactic measure to prevent bacterial colonization of orthopaedic implants and infection. Initial bacterial adhesion onto surgical drill bits as a possible factor for orthopaedic surgical site infections has not yet been documented. The present study investigated the influence of electropolishing on initial staphylococcal adhesion onto AISI 440A stainless steel drill bits. Specifically, one methicillin-susceptible standard laboratory Staphylococcus aureus type strain (DSM 20231T), one methicillin-resistant S. aureus reference strain (DSM 46320) and one methicillin-resistant clinical isolate from an infected orthopaedic implant were used. After standard sterilization, drill bits were immersed in the respective bacterial suspension; bacteria adherent to surface were harvested by vortexing the drill bits in phosphate-buffered saline and viable counts of bacteria transferred from the suspension were made (transferred to log10 for further analysis). Electropolishing significantly reduced adhesion of the clinical S. aureus strain and the S. aureus DSM 20231T. However, electropolishing significantly increased adhesion of the S. aureus DSM 46320. These results show that electropolishing significantly influences initial adhesion of S. aureus strains to surgical drill bits and that the nature of this influence depends on the S. aureus strain examined. For a general recommendation of electropolishing drill bits and guidelines for their handling during surgery, further studies with more strains isolated from infected wounds are suggested.

Course of serum amyloid A (SAA) plasma concentrations in horses undergoing surgery for injuries penetrating synovial structures, an observational clinical study.

BACKGROUND: Injuries penetrating synovial structures are common in equine practice and often result in septic synovitis. Significantly increased plasma levels of serum amyloid A (SAA) have been found in various infectious conditions in horses including wounds and septic arthritis. Plasma SAA levels were found to decrease rapidly once the infectious stimulus was eliminated. The purpose of the current study was to investigate the usefulness of serial measurements of plasma SAA as a monitoring tool for the response to treatment of horses presented with injuries penetrating synovial structures. In the current study plasma SAA concentrations were measured every 48 hours (h) during the course of treatment. RESULTS: A total of 19 horses with a wound penetrating a synovial structure were included in the current study. Horses in Group 1 (n = 12) (injuries older than 24 h) only needed one surgical intervention. Patients in this group had significantly lower median plasma SAA levels (P = 0.001) between 48 h (median 776 mg/L) and 96 h (median 202 mg/L) after surgery. A significant decrease (P = 0.004) in plasma SAA levels was also observed between 96 h after surgery (median 270 mg/L) and 6 days (d) after surgery (median 3 mg/L). Four horses (Group 2) required more than one surgical intervention. In contrast to Group 1 patients in Group 2 had either very high initial plasma concentrations (3378 mg/L), an increase or persistently high concentrations of plasma SAA after the first surgery (median 2525 mg/L). A small group of patients (n = 3) (Group 3) were admitted less than 24 h after sustaining a wound. In this group low SAA values at admission (median 23 mg/L) and peak concentrations at 48 h after surgery (median 1016 mg/L) were observed followed by a decrease in plasma SAA concentration over time. CONCLUSIONS: A decrease in plasma SAA concentrations between two consecutive time points could be associated with positive response to treatment in the current study. Therefore, serial measurements of plasma SAA could potentially be used as an additional inexpensive, quick and easy tool for monitoring the treatment response in otherwise healthy horses presented with injuries penetrating synovial structures. However further studies will be necessary to ascertain its clinical utility.

Algometry to measure pain threshold in the horse's back - An in vivo and in vitro study.

BACKGROUND: The aim of this study was to provide information on algometric transmission of pressure through the dorsal thoracolumbar tissues of the equine back. Using a commercially available algometer, measurements were carried out with six different tips (hemispheric and cylindrical surfaces, contact areas 0.5 cm2, 1 cm2, and 2 cm2). In nine live horses the threshold of pressure that lead to any reaction was documented. In postmortem specimens of five euthanized horses the transmission of algometer pressure onto a pressure sensor placed underneath the dorsal thoracolumbar tissues at the level of the ribs or the transverse lumbar processes respectively was measured. RESULTS: Algometer tips with a contact area of 1 cm2 led to widely similar results irrespective of the surface shape; these measurements also had the lowest variance. Contact areas of 0.5 cm2 resulted in a lower pressure threshold, and those of 2 cm2 resulted in a higher pressure threshold. The hemispheric shape of the contact area resulted in a higher pressure threshold, than the cylindrical contact area. Compared to the thoracic region, a significantly higher pressure threshold was found in the lumbar region in the live horses. This result corresponds to the increased tissue thickness in the lumbar region compared to the thoracic region, also documented as less pressure transmission in the lumbar region on the in vitro specimens. CONCLUSIONS: Algometry is an easily practicable and well tolerated method to quantify pain but it is important to consider the many factors influencing the results obtained.

Novel Methods for Surface EMG Analysis and Exploration Based on Multi-Modal Gaussian Mixture Models.

This paper introduces a new method for data analysis of animal muscle activation during locomotion. It is based on fitting Gaussian mixture models (GMMs) to surface EMG data (sEMG). This approach enables researchers/users to isolate parts of the overall muscle activation within locomotion EMG data. Furthermore, it provides new opportunities for analysis and exploration of sEMG data by using the resulting Gaussian modes as atomic building blocks for a hierarchical clustering. In our experiments, composite peak models representing the general activation pattern per sensor location (one sensor on the long back muscle, three sensors on the gluteus muscle on each body side) were identified per individual for all 14 horses during walk and trot in the present study. Hereby we show the applicability of the method to identify composite peak models, which describe activation of different muscles throughout cycles of locomotion.