Treatment options, complications and long-term outcomes for limb fractures in pet rabbits.

BACKGROUND: Limb fractures represent the most common orthopaedic disease in pet rabbits. However, only a few studies have evaluated therapeutic details of limb fractures. There are no data available for long-term outcomes of limb fracture treatment. METHODS: The medical records of six institutions were reviewed retrospectively to identify cases of traumatic limb bone fractures in pet rabbits between 1999 and 2020. The medical records (n = 387) were analysed for details of fracture prevalence, aetiology, therapy protocols, treatment complications, outcome and long-term effects. In addition to the retrospective data evaluation, 13 rabbits were re-evaluated in person in recent clinical analyses, including orthopaedic examination, radiography and computed-tomographic imaging. Details of long-term effects of fracture treatment were requested over the telephone for a further 232 animals using a standardised questionnaire. RESULTS: Long bone fractures accounted for the majority of all fractures (296/387; 76.5%). Hindlimb fractures (301/387; 77.7%) were more common than forelimb fractures (86/387; 22.2%), and tibial fractures and combined fractures of the tibia and fibula (119/387; 30.8%) were observed most frequently. Most fracture treatments were based on osteosynthesis procedures (243/328; 74.1%). Treatment complications occurred in 130 out of 328 (39.6%) cases. A high bodyweight (p = 0.047) and an older age (p = 0.01) were found to be significant risk factors for the emergence of therapy complications. Overall, 75.4% of animals (175/232) had a satisfactory long-term outcome. Limb posture anomalies were evaluated in 61 cases (26.3%). LIMITATIONS: The multi-centre approach led to the inclusion of various institutions, veterinarians, treatment protocols and rabbit populations that might have influenced the results. The medical records were reviewed retrospectively, so there were some data that were lacking or could not be collected in a standardised manner. Furthermore, rabbit owners' evaluation of long-term outcomes might be prone to error, despite the use of a standardised interview questionnaire. CONCLUSION: Limb fractures are a common orthopaedic issue in pet rabbits. The patient's bodyweight and age are significant risk factors for the emergence of complications during the fracture treatment process. Long-term orthopaedic effects, such as abnormal limb posture and permanent lameness of the affected limb, were observed regularly.

Characterization of microglia/macrophage phenotypes in the spinal cord following intervertebral disc herniation.

Dogs frequently suffer from traumatic spinal cord injury (SCI). Most cases of SCI have a favorable prognosis but 40-50% of dogs with paraplegia and absence of nociception do not regain ambulatory abilities, eventually leading to euthanasia. Microglia and infiltrating macrophages play a crucial role in inflammatory process after SCI. However, little is known about microglia/macrophage phenotypes representing a potential target for future therapeutic strategies. In the present study, the microglia/macrophage phenotype was characterized by immunohistochemistry in the morphologically unaltered canine spinal cord (10 control dogs) and during acute and subacute SCI (1-4 and 5-10 days post injury, 9 and 8 dogs, respectively) using antibodies directed against IBA1, MAC387, MHC-II, lysozyme, EGR2, myeloperoxidase, CD18, CD204 and lectin from Griffonia simplicifolia (BS-1). The expression of these markers was also analyzed in the spleen as reference for the phenotype of histiocytic cells. Histological lesions were absent in controls. In acute SCI, 4 dogs showed mild to moderate hemorrhages, 2 dogs bilateral gray matter necrosis and 6 dogs mild multifocal axonal swellings and myelin sheath dilation. One dog with acute SCI did not show histological alterations except for few dilated myelin sheaths. In subacute SCI, variable numbers of gitter cells, axonal changes and dilated myelin sheaths were present in all dogs and large areas of tissue necrosis in 2 dogs. Neuronal chromatolysis was found in 3 dogs with acute and subacute SCI, respectively. In control dogs, microglia/macrophage constitutively expressed IBA1 and rarely other markers. In acute SCI, a similar marker expression was found except for an increase in MAC387-positive cells in the spinal cord white matter due to an infiltration of few blood-borne macrophages. In subacute SCI, increased numbers of microglia/macrophages expressed CD18, CD204 and MHC-II in the gray matter SCI indicating enhanced antigen recognition, processing and presentation as well as cell migration and phagocytosis during this stage. Interestingly, only CD204-positive cells were upregulated in the white matter, which might be related to gray-white matter heterogeneity of microglia as previously described in humans. The present findings contribute to the understanding of the immunological processes during SCI in a large animal model for human SCI.

In vitro storage of boar spermatozoa increases the demand of adenosine triphosphate for reactivation of motility.

BACKGROUND: Prolonging the shelf-life of liquid-preserved semen without compromising its fertilizing capacity may increase the efficiency of artificial insemination in pigs. Many fertilization-relevant processes are adenosine triphosphate dependent. The impact of semen storage and rewarming to body temperature on the energy status of spermatozoa is as yet unknown. OBJECTIVES: To investigate the energy status of boar spermatozoa during storage and subsequent rewarming and to reveal the potential role of mitochondrial function for reactivation and maintenance of sperm motility. MATERIALS AND METHODS: Extended semen samples (n = 7 boars) were used. Spermatozoa were challenged by storage at 17°C for 7 days and incubation at 38°C for 180 min. The adenosine triphosphate concentration and energy charge in semen samples and lactate concentration in the extracellular medium were assessed. Viability and mitochondrial activity were determined by flow cytometry, and clustered single-cell analysis of motility parameters was performed. RESULTS: The energy status was not affected by semen storage (p > 0.05). Rewarming resulted in a net reduction in adenosine triphosphate concentration, which increased with storage time (maximum Day 5: -24.2 ± 10.3%) but was not accompanied by a loss in viability, motility, or mitochondrial activity. Blocking glycolysis with 2-deoxy-d-glucose prevented the re-establishment of motility and mitochondrial activity after rewarming. Mitochondrial activity gradually subsided in virtually all spermatozoa during incubation at 38°C, while adenosine triphosphate and energy charge remained high. Concomitantly, extracellular lactate levels rose, and sperm populations with lower velocity, increased linearity, and low lateral head displacement grew larger. Size changes for major sperm subpopulations correlated with the percentage of viable spermatozoa with high mitochondrial activity (r = 0.44-0.70 for individual subpopulations, p < 0.01). CONCLUSION: Storage of boar spermatozoa increases the demand of adenosine triphosphate for reactivation of spermatozoa towards fast, non-linear, and hyperactivation-like motility patterns upon rewarming. Maintenance of glycolysis seems to be decisive for sperm function after long-term storage in vitro.

Evaluation of a Treadmill-Based Submaximal Fitness Test in Pugs, and Collecting Breed-Specific Information on Brachycephalic Obstructive Airway Syndrome.

Despite efforts of veterinarians and breeders, brachycephalic obstructive airway syndrome (BOAS) is still a common problem in pugs, underlining the need for objective tests to identify and prevent breeding with affected dogs. In the current study, a submaximal, treadmill-based fitness test was evaluated as a tool to identify signs of airway obstruction not recognisable under rest conditions. In addition to this, different body conformation and measurements were assessed regarding their association with BOAS. A total of 62 pugs and 10 mesocephalic dogs trotted with an individual comfort speed on a treadmill for 15 min. Before and during the examination, dogs were examined for signs of respiratory distress, and a functional BOAS grading was applied. The influence of body conformation on BOAS grading was tested in a univariable and multivariable logistic regression model. During exercise, more respiratory noises were observed, and existing respiratory noises became more apparent in comparison to when at rest. In the multivariable logistic regression model, no factor had a statistically significant influence on BOAS classification. Submaximal fitness testing helped to identify signs of respiratory distress not apparent under resting conditions, and could be a valuable addition for identifying dogs with BOAS. Performing testing on a treadmill facilitates continuous observation of the patients, and enables standardisation of the test regarding the test environment, as well as provides an uninterrupted, steady workload.

Evaluation of blood pressure and NT-proBNP in pugs with and without clinical signs of Brachycephalic Obstructive Airway Syndrome.

Brachycephalic Obstructive Airway Syndrome (BOAS) is a pathologic condition of the upper airways, frequently occurring in dogs of brachycephalic breeds including pugs. It has been suspected that BOAS may be associated with cardiovascular changes and an increased risk for hypertension. The cardiac biomarker NT-proBNP can help to differentiate cardiac from non-cardiac respiratory distress. A possible influence of BOAS on NT-proBNP values has not been investigated, however. The aim of the current study was to examine blood pressure and NT-proBNP levels in pugs with and without clinical signs of BOAS and compare them to values of mesocephalic dogs. For this purpose, NT-proBNP values of 42 pugs and six mesocephalic dogs and blood pressure measurements of 34 pugs and four mesocephalic dogs were explored in the present study. Pugs were examined for clinical signs of BOAS at rest and after a submaximal fitness test, and a functional BOAS grading was applied. Blood pressure (BP) was measured at the beginning and end of the study day and NT-proBNP values were obtained before and after exercise. Measured values of pugs with different degrees of clinical impairment due to BOAS were compared among each other as well as to the CG. In terms of systolic, mean, diastolic BP, and NT-pro BNP, there were no relevant differences between pugs and the CG and no obvious connection between the severity of BOAS symptoms and measured values. BP values of all groups were lower at the second measurement at the end of the study day. NT-proBNP measurements were higher after exercise. BP and NT-proBNP values in all groups were in agreement with commonly used reference ranges. In conclusion, the study adds evidence, that BP and NT-proBNP values did not differ between mesocephalic dogs and pugs with different levels of severity of BOAS but between the measurement times. Thus, in the present study, excitement and exercise seemed to have a greater influence on BP and NT-proBNP values than presence of BOAS symptoms or breed. Discovered values show that the commonly used reference ranges for BP and NT-proBNP are applicable in pugs. This indicates that NT-proBNP can be used to differentiate between cardiac and non-cardiac respiratory distress even in pugs with clinical symptoms of BOAS.

Interactive Rooting Towers and Behavioural Observations as Strategies to Reduce Tail Biting on Conventional Pig Fattening Farms.

Eight pens (25 pigs/pen; n = 200) provided with an interactive straw-filled rooting tower (experimental group) and five pens (25 pigs/pen; n = 125) with a stationary (fixed) tower without straw (control group) were compared within three fattening periods on a conventional farm with fully slatted flooring. The effectiveness of the tower to trigger favourable behaviour in feeding and outside feeding periods was assessed. The incidence of deep tail injuries was lower in the experimental group (experimental group: Odds Ratio 0.3, p < 0.001) and was influenced by the batch (Odds Ratio: 2.38, p < 0.001) but not by pen and sex. In spring, most pens were excluded due to severe tail biting. Tail injury scores were more severe in the control group in weeks 5, 6 and 7 compared to the experimental group (p = 0.002, p < 0.001, p < 0.001, respectively). Tower manipulation was more frequent during feeding compared to outside feeding time (p = 0.002). More head than tail manipulation occurred in the experimental group (p = 0.03). The interactive tower as the only measure was not appropriate to reduce tail biting sufficiently in pigs with intact tails on a conventional fattening farm. Of high priority to prevent tail biting outbreaks was the early detection of biting pigs.

The effect of treatment with pimobendan in dogs with preclinical mitral valve disease - a placebo-controlled double-blinded crossover study.

BACKGROUND: Pimobendan is a widely used medication for the treatment of dogs with congestive heart failure (CHF) and preclinical degenerative mitral valve disease (DMVD) with cardiomegaly. The benefit of a treatment in dogs with preclinical DMVD but without cardiomegaly has not yet been elucidated. Some positive effects concerning life quality and a decrease in cardiac biomarkers could be verified. This study aimed to further investigate these results using a placebo-controlled double-blinded crossover design. Out of a total of 15 dogs, eight were allocated to sequence-group AB, in which dogs received pimobendan (A) during the first treatment period and placebo (B) during the second period. Accordingly, sequence-group BA was treated first with placebo followed by pimobendan. Each treatment period lasted six months and included a baseline investigation and follow-ups after 90 and 180 days. The investigations included a questionnaire completed by the owners, echocardiographic examination, and measurements of NT-proBNP, cTnI and lactate before and after a standardised submaximal exercise test. RESULTS: NT-proBNP values decreased significantly during the treatment period with pimobendan, and the post-exercise increase was attenuated at day 180. No significant treatment effects could be verified for cTnI and lactate, neither pre- nor post-exercise. Left ventricular size decreased under treatment, whereas no significant changes in left atrial size were detected. The owners described their dogs under treatment with pimobendan as being more active at day 90 (11/15) and day 180 (12/15). Those animals treated with placebo were described as being more active at day 90 (2/15) and day 180 (5/15). CONCLUSIONS: Pimobendan had reducing effects on the concentrations of pre- and post-exercise cardiac biomarkers and the size of the left ventricle in dogs with DMVD ACVIM B1. Exercise testing in addition to an assessment of cardiac biomarkers might improve the decision when to initiate pimobendan treatment in dogs with DMVD.

Comparative assessment of left atrial volume in healthy cats by two-dimensional and three-dimensional echocardiography.

BACKGROUND: The left atrium (LA) is an important prognostic parameter in cardiac pathologies of cats. Its size is currently measured in one-dimensional methods, while human medicine considers two- and three-dimensional echocardiography as standard. The objectives of this study were to compare monoplane, biplane, triplane and real-time three dimensional echocardiography for volumetric measurement of the left atrium in healthy cats and establish a reference interval for further studies on cats with heart disease. Additionally, the influence of age, sex and weight on left atrial volume (LAV) was tested. RESULTS: One dimensional monoplane Simpson method of discs (SMOD) in the right parasternal four chamber view (r4) and the left apical 2 chamber view (l2) as well as biplane SMOD had no significant difference for left atrial maximum volume (LAMax). They can be used as equivalent in future studies and one common reference range was set up (1.96 ± 0.54 ml). Those three methods produced significantly higher volumes than triplane echocardiography (RTTPE) and real time three dimensional echocardiography (RT3DE) using TomTec® software. LA volumetry with RTTPE and RT3DE-TomTec™ was more feasible than expected, but low RT3DE image quality was the main reason for excluding patients. Neither age nor weight had an influence on LA volume in healthy cats. Male LAV results were only slightly, but in 2D and RTTPE significantly higher than those of female cats with a range of + 10.46% to + 19.58%. CONCLUSIONS: Monoplane, biplane, triplane and real-time three dimensional echocardiography were feasible for LA volumetry in healthy cats and showed acceptable intra- and interobserver variability. One common LAMax reference range for monoplane r4, l2 and biplane SMOD was set up. Raw data can be used for LA volumes and does not need to be correlated with the cat's weight or age. Male cats have only slightly but significantly larger atria than females in 2D and RTTPE. Therefore, under reservation, also sex related limit values were defined.

Effect of antioxidants, mitochondrial cofactors and omega-3 fatty acids on telomere length and kinematic joint mobility in young and old shepherd dogs - A randomized, blinded and placebo-controlled study.

In dogs, decreasing telomere length is a biomarker for cellular aging. On a systemic level, aging affects the locomotor system in particular, leading to restricted joint mobility. As aging is thought to be related to oxidative stress, it may be counteracted by a diet enriched with antioxidants, mitochondrial cofactors and omega-3 fatty acids. This randomized, blinded and placebo-controlled study examined the influence of an accordingly enriched diet compared to a control diet on 36 young and 38 old shepherd dogs. At the outset, after 3 and after 6 months, mean and minimum telomere lengths were measured. Furthermore, minimum and maximum joint angles and range of motion of the shoulder, elbow, carpal, hip, stifle and tarsal joints were measured by computer-assisted gait analysis. A positive influence of the enriched diet on old dogs could be verified for minimum telomere length and all three parameters of the shoulder joint on the side with the higher vertical ground reaction force after 6 months. In the other joints there were less significant differences; in some cases they indicated a contrary influence of the enriched diet on young dogs, probably due to its reduced protein content. The greater effect of the enriched diet on minimum than on mean telomere length may be due to the higher preference of telomerase for short telomeres. The greater effect on shoulder joint mobility is explained by the greater influence of musculature and connective tissue in this joint. For elderly dogs it is advisable to feed these nutritional supplements.

Influence of lauric acid on the susceptibility of chickens to an experimental Campylobacter jejuni colonisation.

Among the organic acids, lauric acid has shown a high level of in vitro activity against Campylobacter jejuni. The prevalence and intensity of C. jejuni excretion at slaughter often becomes lower with increasing age. In higher-aged broilers on organic farms which often use other breeds, in turn, the prevalence of C. jejuni is sometimes higher at slaughter. The question then arises as to whether a diet with higher lauric acid concentrations, the age alone or the genetic breed might have an effect in the spread and intensity of an experimental C. jejuni infection in vivo. Therefore, two complete diets with or without 2% lauric acid from palm kernel fatty acids were offered to 450 chickens (ten subgroups à 15 birds, repetitions: n = 3) of two broiler and two layer breeds (Ross 308, Hubbard JA 757, Lohmann Dual and Lohmann Brown-Classic). All breeds were reared for 42 days, Lohmann Brown-Classic also for about 98 days. Twenty-one days before dissection, three seeder birds per subgroup were orally infected with a 1 mL inoculum of C. jejuni (4.46±0.35 log10 CFU/mL). Qualitative detection of C. jejuni in cloacal swabs was performed at days 2, 4, 7, 14 after inoculation and at dissection in all birds. Quantitative detection was performed on excreta samples of seeder birds at days 2, 11 and 17 after experimental challenge and on caecal samples of all birds at dissection. Two days after experimental inoculation, C. jejuni prevalence was higher in control birds without lauric acid supplementation (48.9% vs. 39.6%; P = 0.0462). Depending on age, two days after inoculation the C. jejuni prevalence in young Lohmann Brown-Classic chickens was significantly lower (37.8% vs. 61.1%) whereas at dissection it was higher (99% vs. 67%). At day 2 after inoculation C. jejuni counts in the excreta of young Lohmann Brown-Classic were lower in comparison to those in old ones (log10 CFU/g: 3.30±2.68 vs. 5.24±1.56). Eleven (log10 CFU/g: 5.14±1.13 vs. 4.16±0.82) and 17 days after inoculatioin (log10 CFU/g: 3.77±2.02 vs. 1.72±1.87) it was the reverse situation. At dissection, the carriage of C. jejuni in caecal content was higher in younger than in older birds (log10 CFU/g: 8.57±0.46 vs. 6.66±1.43). An effect of genetic breed on C. jejuni prevalence was seen at dissection, this being lowest in Lohmann Dual chickens (91% vs. 98.9% in other breeds). At d 17 after challenge, C. jejuni counts in the excreta of young Lohmann Brown-Classic were lower in comparison to Ross 308 and Hubbard JA 757 (log10 CFU/g: 3.77±2.02 vs. 5.21±0.85 and 5.62±0.90). Lohmann Dual chickens showed an intermediary excretion, this being only significant lower compared to Hubbard JA 757 (log10 CFU/g: 4.31±0.89). In summary, the effect of lauric acid is limited to the initial phase after experimental inoculation. A higher age at infection seems to lead to a more rapid limitation of the infection. The excretion of C. jejuni appears to decrease more rapidly in layer breeds than in broiler lines after experimental inoculation.

Susceptibility of Methicillin-Resistant and -Susceptible Staphylococcus aureus Isolates of Various Clonal Lineages from Germany to Eight Biocides.

Few studies have been conducted on the susceptibility of bacteria to biocides. A total of 182 methicillin-resistant and -susceptible Staphylococcus aureus isolates collected from healthy or diseased humans and animals in Germany were included in the present study. Sixty-three isolates of animal origin and 119 human isolates were tested for their MICs to eight biocides or heavy metals by the broth microdilution method. The MIC50 and MIC90 values of human and animal isolates were equal or differed by not more than 1 dilution step, and statistical analysis revealed that differences between MICs of human and animal isolates were not significant. However, when taking into account the multilocus sequence type (MLST), a strong tendency (P = 0.054) to higher MICs of silver nitrate was detected for clonal complex 398 (CC398) isolates from humans compared to those from animals. Furthermore, a comparison of MIC values from isolates belonging to different clonal lineages revealed that important human lineages such as CC22 and CC5 exhibited significantly (P < 0.05) higher MICs for the biocides chlorhexidine, benzethonium chloride, and acriflavine than the main animal lineage sequence type 398 (ST398). Isolates with elevated MIC values were tested for the presence of biocide and heavy metal tolerance-mediating genes by PCR assays, and the following genes were detected: mepA (n [no. of isolates containing the gene] = 44), lmrS (n = 36), norA (n = 35), sepA (n = 22), mco (n = 5), czrC (n = 3), smr (n = 2), copA (n = 1), qacA and/or -B (n = 1), qacG (n = 2), and qacJ (n = 1). However, only for some compounds was a correlation between the presence of a biocide tolerance gene and the level of MIC values detected.IMPORTANCE Biocides play an essential role in controlling the growth of microorganisms and the dissemination of nosocomial pathogens. In this study, we determined the susceptibility of methicillin-resistant and -susceptible S. aureus isolates from humans and animals to various biocides and heavy metal ions and analyzed differences in susceptibilities between important clonal lineages. In addition, the presence of biocide or heavy metal tolerance-mediating genes was investigated. We demonstrated that important human lineages such as CC22 and CC5 had significantly higher MIC values for chlorhexidine, benzethonium chloride, and acriflavine than the main farm animal lineage, ST398. In addition, it was shown that for some combinations of biocides and tolerance genes, significantly higher MICs were detected for carriers. These findings provide new insights into S. aureus biocide and heavy metal tolerance.

Passive surveillance of Leptospira infection in swine in Germany.

BACKGROUND: As no current data are available on the prevalence of leptospiral infection in swine in Germany, we analysed laboratory data from diagnostic examinations carried out on samples from swine all over Germany from January 2011 to September 2016. A total of 29,829 swine sera were tested by microscopic agglutination test (MAT) for antibodies against strains of eleven Leptospira serovars. RESULTS: Overall, 20.2% (6025) of the total sample collection tested positive for leptospiral infection. Seropositivity ranged between 16.3% (964) in 2011 and 30.9% (941) in 2016 (January to September only). Of all samples, 11.6% (57.3% of the positives) reacted with only one Leptospira serovar, and only 8.6% (42.7% of the positives) reacted simultaneously with two or more serovars. The most frequently detected serovar was Bratislava, which was found in 11.6% (3448) of all samples, followed by the serovars Australis in 7.3% (2185), Icterohaemorrhagiae in 4.0% (1191), Copenhageni in 4.0% (1182), Autumnalis in 3.7% (1054), Canicola in 2.0% (585), and Pomona in 1.2% (368). Modelling shows that both the year and the reason for testing at the laboratory had statistically strong effects on the test results; however, no interactions were determined between those factors. The results support the suggestion that the seropositivities found may be considered to indicate the state of leptospiral infections in the German swine population. CONCLUSION: Although data from passive surveillance are prone to selection bias, stratified analysis by initial reason for examination and analyses by model approaches may correct for biases. A prevalence of about 20% for a leptospiral infection is most probable for sows with reproductive problems in Germany, with an increasing trend. Swine in Germany are probably a reservoir host for serovar Bratislava, but in contrast to other studies not for Pomona and Tarassovi.

The endocannabinoid system in canine Steroid-Responsive Meningitis-Arteritis and Intraspinal Spirocercosis.

Endocannabinoids (ECs) are involved in immunomodulation, neuroprotection and control of inflammation in the central nervous system (CNS). Activation of cannabinoid type 2 receptors (CB2) is known to diminish the release of pro-inflammatory factors and enhance the secretion of anti-inflammatory cytokines. Furthermore, the endocannabinoid 2-arachidonoyl glycerol (2-AG) has been proved to induce the migration of eosinophils in a CB2 receptor-dependent manner in peripheral blood and activate neutrophils independent of CB activation in humans. The aim of the current study was to investigate the influence of the endocannabinoid system in two different CNS inflammatory diseases of the dog, i.e. Steroid-Responsive Meningitis-Arteritis (SRMA) and Intraspinal Spirocercosis (IS). The two main endocannabinoids, anandamide (AEA) and 2-AG, were quantified by mass spectrometry in CSF and serum samples of dogs affected with Steroid- Responsive Meningitis-Arteritis in the acute phase (SRMA A), SRMA under treatment with prednisolone (SRMA Tr), intraspinal Spirocercosis and healthy dogs. Moreover, expression of the CB2 receptor was evaluated in inflammatory lesions of SRMA and IS and compared to healthy controls using immunohistochemistry (IHC). Dogs with SRMA A showed significantly higher concentrations of total AG and AEA in serum in comparison to healthy controls and in CSF compared to SRMA Tr (p<0.05). Furthermore, dogs with IS displayed the highest ECs concentrations in CSF, being significantly higher than in CSF samples of dogs with SRMA A (p<0.05). CSF samples that demonstrated an eosinophilic pleocytosis had the highest levels of ECs, exceeding those with neutrophilic pleocytosis, suggesting that ECs have a major effect on migration of eosinophils in the CSF. Furthermore, CB2 receptor expression was found in glial cells in the spinal cord of healthy dogs, whereas in dogs with SRMA and IS, CB2 was strongly expressed not only in glial cells but also on the cellular surface of infiltrating leukocytes (i.e. neutrophils, eosinophils, lymphocytes, plasma cells, and macrophages) at lesion sites. The present study revealed an upregulated endocannabinoid system in dogs with inflammatory CNS diseases, highlighting the endocannabinoid system as a potential target for treatment of inflammatory CNS diseases.

Utilization and acceptance of virtual patients in veterinary basic sciences - the vetVIP-project.

Context: In medical and veterinary medical education the use of problem-based and cased-based learning has steadily increased over time. At veterinary faculties, this development has mainly been evident in the clinical phase of the veterinary education. Therefore, a consortium of teachers of biochemistry and physiology together with technical and didactical experts launched the EU-funded project "vetVIP", to create and implement veterinary virtual patients and problems for basic science instruction. In this study the implementation and utilization of virtual patients occurred at the veterinary faculties in Budapest, Hannover and Lublin. Methods: This report describes the investigation of the utilization and acceptance of students studying veterinary basic sciences using optional online learning material concurrently to regular biochemistry and physiology didactic instruction. The reaction of students towards this offer of clinical case-based learning in basic sciences was analysed using quantitative and qualitative data. Quantitative data were collected automatically within the chosen software-system CASUS as user-log-files. Responses regarding the quality of the virtual patients were obtained using an online questionnaire. Furthermore, subjective evaluation by authors was performed using a focus group discussion and an online questionnaire. Results: Implementation as well as usage and acceptance varied between the three participating locations. High approval was documented in Hannover and Lublin based upon the high proportion of voluntary students (>70%) using optional virtual patients. However, in Budapest the participation rate was below 1%. Due to utilization, students seem to prefer virtual patients and problems created in their native language and developed at their own university. In addition, the statement that assessment drives learning was supported by the observation that peak utilization was just prior to summative examinations. Conclusion: Veterinary virtual patients in basic sciences can be introduced and used for the presentation of integrative clinical case scenarios. Student post-course comments also supported the conclusion that overall the virtual cases increased their motivation for learning veterinary basic sciences.

Comparative kinematic gait analysis in young and old Beagle dogs.

Age-related involution in dogs involves loss of muscle mass and changes in connective tissue and articular cartilage. The aim of this study was to examine whether an age-related influence on joint mobility can be detected in the absence of disease. Five young (mean age 2.0 years) and five old (mean age 10.4 years) healthy and sound Beagle dogs underwent computer-assisted gait analysis during locomotion on a treadmill. Shoulder, elbow, carpal, hip, stifle, and tarsal joint angles including joint angle progression curves, minimum and maximum joint angles, and range of motion (ROM) in degrees were analyzed. The old group had a smaller maximum joint angle (p = 0.037) and ROM (p = 0.037) of the carpal joint; there were similar tendencies in the shoulder, elbow, and carpal joints. Descriptive analysis of the progression curves revealed less flexion and extension of the forelimb joints. The results indicate restricted joint mobility of the forelimb in old dogs, primarily of the carpal joint. Results in the joints of the hindlimb were inconsistent, and the contrasting alterations may be due to a compensatory mechanism. As most alterations were found in the distal joints, these should receive particular attention when examining elderly dogs.

Th17-skewed immune response and cluster of differentiation 40 ligand expression in canine steroid-responsive meningitis-arteritis, a large animal model for neutrophilic meningitis.

BACKGROUND: Steroid-responsive meningitis-arteritis (SRMA) is an immune-mediated disorder characterized by neutrophilic pleocytosis and an arteritis particularly in the cervical leptomeninges. Previous studies of the disease have shown increased levels of IL-6 and TGF-ß1 in cerebrospinal fluid (CSF). In the presence of these cytokines, naive CD4+ cells differentiate into Th17 lymphocytes which synthesize interleukin 17 (IL-17). It has been shown that IL-17 plays an active role in autoimmune diseases, it induces and mediates inflammatory responses and has an important role in recruitment of neutrophils. The hypothesis of a Th17-skewed immune response in SRMA should be supported by evaluating IL-17 and CD40L, inducing the vasculitis. METHODS: An enzyme-linked immunosorbent assay (ELISA) was performed to measure IL-17 and CD40L in serum and CSF from a total of 79 dogs. Measurements of patients suffering from SRMA in the acute state (SRMA A) were compared with levels of patients under treatment with steroids (SRMA T), recurrence of the disease (SRMA R), other neurological disorders, and healthy dogs, using the two-part test. Additionally, secretion of IL-17 and interferon gamma (IFN-γ) from the peripheral blood mononuclear cells (PBMCs) was confirmed by an enzyme-linked immunospot (ELISpot) assay. RESULTS: Significant higher levels of IL-17 were found in CSF of dogs with SRMA A compared with SRMA T, other neurological disorders and healthy dogs (p < 0.0001). In addition, levels of CD40L in CSF in dogs with SRMA A and SRMA R were significantly higher than in those with SRMA T (p = 0.0004) and healthy controls (p = 0.014). Furthermore, CSF concentrations of IL-17 and CD40L showed a strong positive correlation among each other (rSpear = 0.6601; p < 0.0001) and with the degree of pleocytosis (rSpear = 0.8842; p < 0.0001 and rSpear = 0.6649; p < 0.0001, respectively). IL-17 synthesis from PBMCs in SRMA patients was confirmed; however, IL-17 is mainly intrathecally produced. CONCLUSIONS: These results imply that Th17 cells are inducing the autoimmune response in SRMA and are involved in the severe neutrophilic pleocytosis and disruption of the blood-brain barrier (BBB). CD-40L intrathecal synthesis might be involved in the striking vasculitis. The investigation of the role of IL-17 in SRMA might elucidate important pathomechanism and open new therapeutic strategies.

Infection Dynamics and Antimicrobial Resistance Profile of Salmonella Paratyphi B d-tartrate Positive (Java) in a Persistently Infected Broiler Barn.

The infection dynamics of S. Java were examined in three consecutive rearing periods on a broiler farm in Northwestern Germany which had been persistently infected with S. Java for more than five years. The barn was investigated for Salmonella occurrence after cleaning and disinfection to verify the persistent contamination of the broiler house with S. Java before the start of the first rearing cycle. Confirmation of Salmonella absence in day-old chicks (time-point 1) as well as early establishment of infection between days 5-7 (time-point 2) were confirmed by caecal swabs prepared for qPCR and classical microbiological methods. At three time-periods (between days 11-15 (time-point 3), days 25-28 (time-point 4), and days 38-40 (time-point 5)) caecal content was examined for colony forming units (CFU) Salmonella/g. In general, there was an increase in Salmonella Java load at time-point 4 compared to time-points 3 and 5. Therefore, we observed a bell-shaped course of infection resulting in higher rates of Salmonella CFU/g prior to prethinning than at final slaughter. The antimicrobial susceptibility testing revealed resistance to tetracycline, fluorquinolones, trimethoprim, and cefoxitin.

Towards a Standardized Method for Broth Microdilution Susceptibility Testing of Haemophilus parasuis.

Currently, there is no agreed method available for broth microdilution susceptibility testing of Haemophilus parasuis, one of the most important bacterial pathogens in pig production. Therefore, the aim of this study was to develop a method that could be easily performed by diagnostic laboratories and that appears suitable for a harmonized susceptibility testing. Growth determinations using one type strain and three field isolates revealed no visible growth of H. parasuis in media which have proven to be suitable for susceptibility testing of fastidious organisms. Therefore, a new medium, cation-adjusted Mueller-Hinton broth (CAMHB) plus NADH and sterile filtered heat-inactivated chicken serum, was developed. The reproducibility of MICs obtained in this medium was evaluated and statistically analyzed, considering a model with two different variables (precondition of five identical MICs and MIC mode accepting a deviation of ±1 dilution step, respectively). No significant differences for both variables were seen between two time points investigated and between results obtained with the recently proposed test medium broth (TMB). Nearly all MICs of quality control strains were in the acceptable range. Subsequently, 47 H. parasuis isolates representing 13 serovars were tested with the newly developed medium and TMB. Statistical analysis of all isolates and 15 antimicrobial agents and antimicrobial combinations showed no significant difference between MICs obtained in supplemented CAMHB and TMB. Because of a simplified implementation in routine diagnostic and a lower chance of interference between medium components and antimicrobial agents, supplemented CAMHB is recommended with an incubation time of 24 h.

Oral Supplementation with a Special Additive of Retinyl Palmitate and Alpha Tocopherol Reduces Growth Retardation in Young Pancreatic Duct Ligated Pigs Used as a Model for Children Suffering from Exocrine Pancreatic Insufficiency.

Pancreatic exocrine insufficiency (PEI) is a disease of diverse aetiology-e.g., majority of patients suffering from cystic fibrosis (CF) show PEI congenitally. Malnutrition and malabsorption of nutrients impair growth and nutritional status. As reduced fat digestion leads to a deficiency of fat-soluble vitamins the supplementation is standard, but absorption is a critical point in PEI-patients. The pancreatic duct ligated (PL) pig is an established model for PEI in humans and has been proven to be a suitable model to compare different vitamin additives for supplementation. In a former study, PEI caused distinct growth retardation in young piglets, but did not affect growth in older ones. Our study hypothesised that this age-dependent effect is caused by exhausted body reserves of fat-soluble vitamins and, therefore, extra supply reduces growth retardation. PEI was induced by PL at the age of seven (PL-7) or 16 weeks (PL-16). Controls (C) underwent a sham surgery. Some PL-7 pigs (PL-7 + Vit) were fed a special vitamin additive. PEI reduced the mean final body weight (kg) at 26 weeks of age significantly with lower effect in PL-16-pigs (C:117; PL-7:49.5; PL-7 + Vit:77.1; PL-16:96.4). Extra vitamin supply resulted in an increased growth and normalised serum concentration of alpha-tocopherol, underlining the importance of special supplementation in PEI-patients.

Evaluation of Functionalized Porous Titanium Implants for Enhancing Angiogenesis in Vitro.

Implant constructs supporting angiogenesis are favorable for treating critically-sized bone defects, as ingrowth of capillaries towards the center of large defects is often insufficient. Consequently, the insufficient nutritional supply of these regions leads to impaired bone healing. Implants with specially designed angiogenic supporting geometry and functionalized with proangiogenic cytokines can enhance angiogenesis. In this study, Vascular Endothelial Growth Factor (VEGF) and High Mobility Group Box 1 (HMGB1) were used for incorporation into poly-ε-caprolactone (PCL)-coated porous titanium implants. Bioactivity of released factors and influence on angiogenesis of functionalized implants were evaluated using a migration assay and angiogenesis assays. Both implants released angiogenic factors, inducing migration of endothelial cells. Also, VEGF-functionalized PCL-coated titanium implants enhanced angiogenesis in vitro. Both factors were rapidly released in high doses from the implant coating during the first 72 h.