Comparison of bone healing,as assessed by computed tomography,following tibial tuberosity advancement in dogs with and without autogenous cancellous bone grafts

Abstract

AIMS: To objectively compare measures of bone healing, using computed tomography (CT) in dogs following bilateral tibial tuberosity advancement (TTA), between tibiae treated with and without autogenous cancellous bone grafts.

METHODS: Ten dogs with bilateral cranial cruciate ligament disease requiring surgical stabilisation were prospectively recruited to undergo single-session bilateral TTA, with only one, randomly assigned, tibia receiving bone graft in the osteotomy deficit. Bone healing at the osteotomy site was assessed using CT performed 38–70 days post-operatively. CT images were evaluated using both objective measurements of osseous bridging and subjective evaluation by six radiologists. Repeated measures ANOVA was used to compare the objective outcomes between the grafted and non-grafted tibiae.

RESULTS: The mean percentage of the osteotomy deficit bridged at the lateral cortex was greater in grafted (77.6, SD 35.2%) compared to non-grafted (63.0, SD 36.5%) tibiae (p=0.001), but did not differ at the medial cortex (p=0.1). The mean minimum callus width was greater in grafted (7.2, SD 3.3 mm) compared to non-grafted (3.6, SD 2.9 mm) tibiae (p<0.001). There was no difference in mean attenuation (measured in Hounsfield units) of the callus between grafted and non-grafted tibiae (p=0.5). The grafted tibia was deemed to have superior bone healing in 50/60 subjective assessments made by radiologists.

CONCLUSIONS: Superior osseous bridging was detected by CT analysis following TTA using autogenous cancellous bone grafts compared with no graft. This was shown by greater bridging percentage at the lateral cortex and formation of a broader callus. Qualitative assessments made by six radiologists also supported the conclusion that bone healing was improved by use of autogenous cancellous bone graft. CT was a useful method for assessing evidence of bone healing following TTA.

CLINICAL RELEVANCE: These findings justify the application of autogenous cancellous bone graft to augment healing following TTA in dogs.     

Prolactin,Androstenedione and IGF1 Serum Concentrations During Induced Follicular Growth by eCG Administration in the Bitch

The oestrus cycle in the domestic bitch, a monoestrous species, differs considerably from that of other veterinary domestic animals species. In the bitch the combined use of eCG and hCG is effective to induce oestrus predictably and safely (Stornelli et al., Theriogenology, 78, 2012 and 1056). Although several studies were done to describe the hormonal changes during the canine oestrus cycle, to our knowledge none was done to describe the hormonal changes during induced follicular growth after the administration of eCG. The aim of this work was to study prolactin (PRL), insulin‐like growth factor (IGF1) and androstenedione (ANDR) serum concentrations during follicular growth induced by a single dose of eCG administered to late anoestrous bitches. PRL and ANDR concentrations were lower before than after eCG TRT (before eCG vs pro‐oestrus, oestrus and dioestrus; 4.3 ± 1.8 ng/ml vs 6.5 ± 1.6 ng/ml, p < 0.05; 0.08 ± 0.2 ng/ml vs 0.42 ± 0.16 ng/ml, p < 0.05). Conversely, IGF1 concentrations were similar before and after eCG TRT (286.0 ng/ml ±32.2, p > 0.53). Additionally, PRL concentrations were similar before oestrus compared to during oestrus and dioestrus (6.9 ± 1.7 ng/ml, p > 0.19). Furthermore, IGF1 concentrations were higher before and during oestrus compared to first day of dioestrus (286.1 ± 29.8vs 200.4 ± 29.2 ng/ml, p < 0.01). On the contrary, ANDR concentrations were lower before and during oestrus compared to first day of diestrum (0.35 ± 0.17 ng/ml and 0.38 ± 0.15 vs 0.68 ± 0.17 ng/ml, p < 0.05). These results show that treatment with a single injection of 50 IU/kg of eCG in late anoestrous bitches successfully induced changes in follicular growth which were paralleled with changes in PRL, IGF1 and ANDR serum concentration similar to those occurring during a normally occurring oestrous cycle. In addition, our results suggest that IGF1 in the bitch could play an important role in ovarian folliculogenesis.     

Chlorophyll fluorescence as an indicator of frost hardiness in white spruce seedlings from different latitudes

This study examined the utility of variable chlorophyll fluorescence (F_var) to detect freezing damage in white spruce seedlings of four seedlots. Logistic regression analysis done for freezing tests in September showed that visible needle damage from freezing could be estimated by the F_var attributes F_o/I_ABS(r²=0.94), F_p(r²=0.98), F_v/F_m (r²=0.99), and F₁(r²=0.86). The regression curves indicated that for all four fluorescence attributes, inflection points occurred between 10 and 20% visible needle damage. The lack of a relationship between fluorescence attributes and visible seedling needle damage in October through December is because the minimum temperature (–18 and –24°C respectively) applied was insufficient to cause needle damage. Freezing-induced changes to F_var attributes can be detected which also result in photosynthetic rate decreases when no visible needle damage, and even electrolyte conductivity changes are evident. F_var attribute differences due to freezing can be resolved to the seedlot level. The Fvar curve feature manifested 5 seconds after dark-adapted seedlings have been exposed to light (F_5s) will estimate (r²=0.76) photosynthetic rate after freezing.     

Overwinter Storability of Conifer Planting Stock: Operational Testing of Fall Frost Hardiness

Operational stock-testing facilities that estimate overwinter storability of seedlings (ability to survive and grow after storage) need a reliable method that provides fast results to forest nurseries. We compared three methods using container-grown seedlings of Douglas-fir, interior spruce, lodgepole pine, and western larch from forest nurseries in British Columbia. On three to nine dates in autumn, frost hardiness at −18°C was estimated using visible injury of foliage or stems (VI), electrolyte leakage from needles or stems (EL), and chlorophyll fluorescence of shoots (CF). Seedlings were placed into overwinter cold storage (−2°C). In the spring, stored seedlings were planted in nursery beds; survival and growth were assessed after one growing season. There were close correlations (r ≥ 0.93) between the assessment methods. Seedlings lifted after they reached thresholds of 69% or higher for CF and 25% or lower for EL and VI had over 90% survival at harvest and doubled shoot dry weight compared with seedlings lifted earlier. Measuring CF was the fastest and most easily replicated method to estimate successful storability, and reduced testing time by 6 days relative to VI tests.     

Seasonal changes in chlorophyll fluorescence of white spruce seedlings from different latitudes in relation to gas exchange and winter storability

Variable chlorophyll fluorescence (F_var) was investigated as a tool in detection of distinct seasonal physiological changes in 1+0 intact white spruce seedlings. The loss of the characteristic F_var peak (Fp) between 0.8 and 1.0 s after illumination of dark adapted seedlings is an indication of regulation of photosynthetic activity in August. The peak represents excess photochemical water-splitting of photosystem II. We interpret its loss as a physiological indicator of the process of dormancy induction. Three dimensional (i.e. X[0-300 s], Y[rfu], Z[time of year] axis) F_var curves of non-stressed seedlings measured over 300 s followed a three phase change over the growing season. In actively growing seedlings, the portion of the Kautsky induction curve between 60 and 300 s was 0.4 relative fluorescence units (rfu) in northern (i.e. >56° latitude) seedlots and 0.6 in the southern (i.e. <56° latitude) range seedlot from August until early September. About mid-September curve features between 60 and 300 s decreased sharply to approximately zero (rhu) by October. Freeze test data indicated seedlings became frost hardy during this time. The third, or inactive phase was seen as flat line from 40 to 100 s. The portion of the curve after 100 s was responsive to short term temperature changes. White spruce seedlots of northern and southern B.C. latitudes having curve fluorescence peak (F_p) values at about 1 s of 0.6, and 0.8 (rhu) respectively, plus curve minimum (F_min) values at about 60 s which do not decrease further over a 2–3 week period represent stock which can safely be lifted for cold storage. The F_var attribute at 5 s after the actinic light is turned on (F_5s) correlates well with net photosynthesis (r² =0.61) during the growing season.     

Hazard Analysis and Critical Control Points System for a Bull Semen Production Centre

Bull semen production centres (SPC) generally present satisfactory quality control for sperm processing, but non‐standardized hygiene procedures. This study describes a Hazard Analysis and Critical Control Points (HACCP) system developed for bull SPC and subsequently implemented in a commercial SPC. After the identification of hazards at each step of semen processing and the determination of their risk and severity, monitoring and corrective procedures were designed to assess the system's efficiency. The HACCP system identified six microbiological hazards, 10 physical hazards, four chemical hazards and three critical control points. After the establishment of Good Processing Practices, Standard Operating Procedures and Standard Sanitizing Operating Procedures, the system was validated through an audit, to identify eventual failures and to define measures to correct them.     

Fate of Maize DNA During Steeping,Wet-Milling,and Processing

The fate of DNA during steeping, wet-milling, and subsequent processing of maize was examined using a sensitive polymerase chain reaction (PCR-based) detection system. The system used specific amplification of maize DNA sequences by primers generated toward plant nuclear- and chloroplast-encoded genes. The PCR method facilitated analysis of DNA content in food products, which is an important issue in use of genetically modified organisms. In a conventional laboratory wet-milling countercurrent steep system, DNA was detected in maize kernels throughout the process but was not found in steepwater. After kernels were wet-milled, DNA was detected in the starch, germ, coarse fiber, and wet gluten fractions but not in the fine fiber fraction. When dried by heating at 135°C for 2 hr, DNA was degraded to undetectable levels in the wet-milled gluten fraction and hydrated kernels. DNA was not detected in feed pellets, starch, dextrose, sorbitol, or high-fructose maize syrup made from industrial wet-milled samples. Although DNA could be detected in laboratory wet-milled fractions, some degree of degradation occurred after extended exposure to steepwater. Countercurrent steepwater samples from the later stages of the steeping process were able to degrade DNA. The level of DNA degradation appeared to correspond to the presence of sulfur dioxide and may represent a physiochemical rather than an enzyme-mediated process. Our results indicate that some steps in the steeping and wet-milling process can degrade maize genomic and plastid DNA.     

Molecular detection of WA‐CMS restorers from tropical japonica‐derived lines,their evaluation for fertility restoration and adaptation

Hybrid rice based on wild‐abortive cytoplasmic male sterility (WA‐CMS) is important in boosting rice production, which requires diverse parents to harness heterosis. For this, exploiting the diversity of japonica through tropical japonica (TRJ) lines is an excellent route. In this study, 310 TRJ‐based new plant type (NPT) lines were developed and evaluated for Rf3 and Rf4 genes. Gene‐based (DRRM‐Rf3‐5 and DRRM‐Rf3‐10) and functional marker (RMS‐SF21‐5) targeted Rf3 locus, while gene‐linked (RM6100) and functional marker (RMS‐PPR9‐1) targeted the Rf4 locus. The frequency of the restorer allele of Rf3 gene was lower when compared to that of Rf4. Combined phenotypic and molecular screening using gene‐based and functional markers identified 42 lines that carried Rf3 and/or Rf4 genes. All the selected lines produced fertile F₁s when crossed to a WA‐CMS line, “Pusa 6A”, but with varying levels of spikelet fertility. This is the first report of a marker‐cum‐phenotype‐based restorer selection using TRJ‐derived lines. Multilocation evaluation of these lines at three locations indicated better adaptation for grain yield in some of the lines.     

Concurrent Aelurostrongylus abstrusus infection and salmonellosis in a kitten

A 14-week-old kitten had a history of vomiting, diarrhoea and pyrexia, all of which resolved without treatment. Three weeks later the kitten developed a violent non-productive dry cough. Thoracic radiographs revealed pneumothorax and nodular alveolar disease. Aelurostrongylus abstrusus larvae and intracellular Gram-negative bacilli were seen in bronchial wash fluid and pleural exudate, and Salmonella Typhimurium was cultured from both fluids but not from faeces. Therapy included unilateral closed-tube thoracostomy, enrofloxacin and fenbendazole. Historical signs were compatible with gastrointestinal salmonellosis and secondary broncho-pneumonia. Seeding of the lungs with salmonellae may have occurred as a result of migration of A abstrusus from a gastro-intestinal tract residually infected or colonised by S Typhimurium. Alternatively, the development of lungworm infection in the cat may have activated quiescent S Typhimurium pulmonary granulomata from bacteraemia secondary to gastro-intestinal salmonellosis. Two years after diagnosis the cat was reportedly in good health.