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

Comparison of Selective Laser Melted Titanium and Magnesium Implants Coated with PCL

Degradable implant material for bone remodeling that corresponds to the physiological stability of bone has still not been developed. Promising degradable materials with good mechanical properties are magnesium and magnesium alloys. However, excessive gas production due to corrosion can lower the biocompatibility. In the present study we used the polymer coating polycaprolactone (PCL), intended to lower the corrosion rate of magnesium. Additionally, improvement of implant geometry can increase bone remodeling. Porous structures are known to support vessel ingrowth and thus increase osseointegration. With the selective laser melting (SLM) process, defined open porous structures can be created. Recently, highly reactive magnesium has also been processed by SLM. We performed studies with a flat magnesium layer and with porous magnesium implants coated with polymers. The SLM produced magnesium was compared with the titanium alloy TiAl6V4, as titanium is already established for the SLM-process. For testing the biocompatibility, we used primary murine osteoblasts. Results showed a reduced corrosion rate and good biocompatibility of the SLM produced magnesium with PCL coating.DFG/299/11-

SLM produced porous titanium implant improvements for enhanced vascularization and osteoblast seeding

To improve well-known titanium implants, pores can be used for increasing bone formation and close bone-implant interface. Selective Laser Melting (SLM) enables the production of any geometry and was used for implant production with 250-microm pore size. The used pore size supports vessel ingrowth, as bone formation is strongly dependent on fast vascularization. Additionally, proangiogenic factors promote implant vascularization. To functionalize the titanium with proangiogenic factors, polycaprolactone (PCL) coating can be used. The following proangiogenic factors were examined: vascular endothelial growth factor (VEGF), high mobility group box 1 (HMGB1) and chemokine (C-X-C motif) ligand 12 (CXCL12). As different surfaces lead to different cell reactions, titanium and PCL coating were compared. The growing into the porous titanium structure of primary osteoblasts was examined by cross sections. Primary osteoblasts seeded on the different surfaces were compared using Live Cell Imaging (LCI). Cross sections showed cells had proliferated, but not migrated after seven days. Although the cell count was lower on titanium PCL implants in LCI, the cell count and cell spreading area development showed promising results for titanium PCL implants. HMGB1 showed the highest migration capacity for stimulating the endothelial cell line. Future perspective would be the incorporation of HMGB1 into PCL polymer for the realization of a slow factor release.DFG/299/11-

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

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-beta(1) 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-gamma) 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

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

Starch Origin and Thermal Processing Affect Starch Digestion in a Minipig Model of Pancreatic Exocrine Insufficiency

Although steatorrhea is the most obvious symptom of pancreatic exocrine insufficiency (PEI), enzymatic digestion of protein and starch is also impaired. Low praecaecal digestibility of starch causes a forced microbial fermentation accounting for energy losses and meteorism. To optimise dietetic measures, knowledge of praecaecal digestibility of starch is needed but such information from PEI patients is rare. Minipigs fitted with an ileocaecal fistula with (n=3) or without (n=3) pancreatic duct ligation (PL) were used to estimate the rate of praecaecal disappearance (pcD) of starch. Different botanical sources of starch (rice, amaranth, potato, and pea) were fed either raw or cooked. In the controls (C), there was an almost complete pcD (>92%) except for potato starch (61.5%) which was significantly lower. In PL pcD of raw starch was significantly lower for all sources of starch except for amaranth (87.9%). Thermal processing increased pcD in PL, reaching values of C for starch from rice, potato, and pea. This study clearly underlines the need for precise specification of starch used for patients with specific dietetic needs like PEI. Data should be generated in suitable animal models or patients as tests in healthy individuals would not have given similar conclusions

Quantitative morphological changes in the interplacentomal wall of the gravid uterine horn of cattle during pregnancy

Background: The interplacentomal wall of the gravid uterine horn in cattle is the subject of reports dealing mainly with specific aspects of early pregnancy or the peripartal period. Only a very limited number of early and descriptive studies includes the whole period of pregnancy. Thus, there is a gap concerning quantitative morphological data of the uterine wall during pregnancy. We hypothesized that the specific requirements of pregnancy are reflected by significant and characteristic morphologic changes. Methods: Interplacentomal segments of the fetus-bearing horn of the uterus of 47 cows were collected at slaughter, assessed quantitatively by light microscopy, grouped into trimesters (trim), and data were analyzed statistically. Results: During pregnancy there were significant increases (p<0.05) in the measured parameters: heights of the endometrial surface epithelium (31 increased to 46 and 46 μm, in the 1st, 2nd and 3rd trim, respectively), glandular epithelium (19.6 to 22.4 and 25.4 μm, respectively), diameters of glands (94 to 166 to 239 μm, respectively) and glandular lumina (56 to 122 to 188 μm, respectively). Volume density of the glandular epithelium did not change, while that of glandular lumina increased significantly (8 to 26 to 40% in the 1st, 2nd and 3rd trim, respectively) and of endometrial stroma decreased with ongoing pregnancy (67 to 46 to 37%; p<0.05). Diameters of myometrial smooth muscle cells (MSMC) (9.7 to 12.4 and 12.9 μm, respectively, for the 1st, 2nd and 3rd trim; p<0.05), and the volume fraction of myometrial stroma increased (6 to 10 to 13%; p<0.05), while decreases were observed in MSMC nuclear volume density (4.4 and 4.0 to 2.4%; p<0.05). The fraction of MSMC cytoplasm (89 to 85%) and the nucleus:cytoplasm ratio (0.05 to 0.03%) both decreased for the 1st vs. 3rd trim, respectively (p<0.05). Conclusions: These results indicate that the interplacentomal wall of the gravid uterine horn is subjected to significant morphological changes during pregnancy, underlining the importance of endometrial surface epithelium and of gland hypertrophy for nourishment of the conceptus, of increased myometrial extracellular matrix for uterine tensile strength and of myometrial smooth muscle hypertrophy for expulsion of the fetus at term