A Novel Approach to the Treatment and Prevention of Laminitis: Botulinum Toxin Type A for the Treatment of Laminitis

Laminitis is an inflammation of the lamina of hoofed animals. According to the United States Department of Agriculture (USDA), laminitis impacts approximately 2% of the horse population each year. Because of the severity of the inflammation and chronic pain, it is frequently necessary to euthanize these horses. Surviving horses may be left useless, with resultant economic and social impact on the industry. Current interventions for laminitis are based on supportive care and alteration of biomechanical forces on the digit. The transition from laminitis to laminar failure (founder) is believed to be caused by the “weight of the horse and the forces of locomotion driving the bone down into the hoof capsule.” We hypothesize that an important factor is the torsional forces placed on the distal phalanx (coffin bone) by the deep digital flexor muscle and tendon. The paralyzing effect of botulinum toxin will result in a decrease of these torsional forces and therefore will aid in the prevention or treatment of the sequelae of laminitis. Seven horses with varying degrees of laminitis were evaluated. Each underwent pretreatment and posttreatment radiographs as well as pretreatment and posttreatment Obel grading. Each horse received botulinum toxin type A injected into the belly of the deep digital flexor muscle. In all cases there was radiographic stabilization of pedal displacement from the dorsal hoof wall. Obel scores showed improvement of 1 to 2 grades during the time monitored.     

The hen's egg: Shell cracking in eggs dropped on cage floors coated with plastic or zinc

A sequential method of estimating the mean and variance or the drop height (h ₅₀) at which the shells of 50% of eggs crack when dropped on a cage floor was used at each of four locations on each of five cage floors, made of 2.05 mm diameter (14 SWG) steel wire, that were similar save in respect of the coating on the wire. The coatings were: none, zinc at 2.7 g/m and polyvinyl chloride (PVC) at 2.0, 3.0 or 6.6 g/m. Relative to the uncoated floor the zinc‐coated one was markedly worse in respect of shell cracking and the PVC‐coated floors were marginally better. The relationship of h ₅₀ to the weight of PVG coating was curvilinear, with an optimum at about 4 g/m. The method of estimation proved very efficient.     

Survival in Dogs with Nasal Adenocarcinoma: 64 Cases (1981–1995)

Case records of 64 dogs with nasal adenocarcinoma were reviewed. The effects of age, gender, tumor stage, presence of metastatic lesions, and treatment method on survival time were examined. Surgery groups included rhinotomy (n = 9), transnasal curettage (n = 29), and no surgery (n = 26). Chemotherapy groups included fluorouracil-cyclophosphamide combination therapy (n = 15), mitoxantrone (n = 7), and no chemotherapy (n = 42). Fifty-three dogs received fractionated cobalt 60 radiation therapy. Surgical procedure, chemotherapy group, and stage of primary tumor were not significantly associated with survival time ( P > .05). Dogs that received radiation therapy had a significantly longer median survival time (424 days) than dogs that did not (126 days) ( P = .0001). The presence of either regional lymph node or pulmonary metastasis was associated with significantly shorter median survival time (109 days) when compared to dogs without metastases (393 days) ( P = .0125). When only dogs that had received radiation therapy were considered, neither surgical treatment nor chemotherapy group was associated with significant changes in median survival time. An alternate staging system emphasizing the presence or absence of metastases is proposed. Key words: Chemotherapy; Metastasis; Radiotherapy.     

Presettlement and modern disturbance regimes in coast redwood forests: Implications for the conservation of old-growth stands

Coast redwood (Sequoia sempervirens), a western North American conifer of ancient lineage, has a paradoxical combination of late-successional characteristics and strong adaptations to disturbance. Despite its shade tolerance and heavy dominance of the canopy on many sites, redwood saplings are uncommon in upland old-growth stands. Information needed to ensure the conservation of old-growth redwood forests has been limited. In this review paper, we integrate evidence on redwood biology with data on the historic and modern disturbance regimes to help clarify the degree to which key attributes of redwood forests may have been dependent upon periodic disturbance. Available evidence suggests that episodes of fire, flooding, and slope failure prior to European settlement were frequent but predominantly of low to moderate severity and extent, resulting in broadly uneven-aged forests. The majority of fires prior to European settlement were apparently of human origin. Frequency and severity of the major disturbance agents have been radically changed in modern times. Fires have been largely excluded, and flooding has been altered in ways that have often been detrimental to old-growth redwoods on alluvial terraces. However, because of the apparent anthropogenic origin of most presettlement fires, the long-term evolutionary role of fire for coast redwood is ecologically ambiguous. With fire exclusion, redwood possibly could be displaced to some extent on upland sites by increasing abundance of fire-sensitive competitors. Alternatively, redwood may be able to maintain dominance by vegetative sprouting and new seedling establishment on root-wad mounds, fallen logs, and on soil exposed by slope failure. Future research priorities are suggested that will help resolve some of the current ambiguities.     

Canine corneal fibroblast and myofibroblast transduction with AAV5

Objective The aims of this study were (1) to determine the efficacy of adeno‐associated vector serotype 5 (AAV5) for delivering gene therapy to canine corneal fibroblasts (CCFs) and myofibroblasts (CCMs) using enhanced green fluorescent protein (GFP) marker gene and (2) to evaluate the cytotoxicity of AAV5 to CCFs and CCMs using an in vitro model. Methods Healthy donor canine corneas were used to generate primary CCFs by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum‐free medium containing transforming growth factor β1 (1 ng/mL). An AAV5 titer (6.5 × 10¹² μg/mL) expressing GFP under control of hybrid cytomegalovirus + chicken β‐actin promoters (AAV5‐gfp) was used to transduce CCF and CCM cultures. Delivered gene expression in CCFs and CCMs was quantified using immunocytochemistry, fluorescent microscopy, and real‐time PCR. Transduction efficacy of the AAV5 vector was determined by counting DAPI‐stained nuclei and EGFP‐positive cells in culture. Phase‐contrast microscopy, trypan blue, and dUTP nick end labeling (TUNEL) assays were used to determine the toxicity and safety of AAV5 in this canine corneal model. Results Topical AAV5 application successfully transduced a significant population of CCFs (42.8%; P < 0.01) and CCMs (28%; P < 0.01). Tested AAV5 did not affect CCF or CCM phenotype or cellular viability and did not cause significant cell death. Conclusions The tested AAV5 is an effective and safe vector for canine corneal gene therapy in this in vitro model. In vivo studies are warranted.     

Effect of pharmacological concentrations of zinc oxide with or without the inclusion of an antibacterial agent on nursery pig performance

A study involving nine research stations from the NCR-42 Swine Nutrition Committee used a total of 1,978 crossbred pigs to evaluate the effects of dietary ZnO concentrations with or without an antibacterial agent on postweaning pig performance. In Exp. 1, seven stations (IA, MI, MN, MO, NE, ND, and OH) evaluated the efficacy of ZnO when fed to nursery pigs at 0, 500, 1,000, 2,000, or 3,000 mg Zn/kg for a 28-d postweaning period. A randomized complete block experiment was conducted in 24 replicates using a total of 1,060 pigs. Pigs were bled at the 28-d period and plasma was analyzed for Zn and Cu. Because two stations weaned pigs at < 15 d (six replicates) and five stations at > 20 d (18 replicates) of age, the two sets of data were analyzed separately. The early-weaned pig group had greater (P < 0.05) gains, feed intakes, and gain:feed ratios for the 28-d postweaning period as dietary ZnO concentration increased. Later-weaned pigs also had increased (P < 0.01) gains and feed intakes as the dietary ZnO concentration increased. Responses for both weanling pig groups seemed to reach a plateau at 2,000 mg Zn/kg. Plasma Zn concentrations quadratically increased (P < 0.01) and plasma Cu concentrations quadratically decreased (P < 0.01) when ZnO concentrations were > 1,000 mg Zn/kg. Experiment 2 was conducted at seven stations (KY, MI, MO, NE, ND, OH, and OK) and evaluated the efficacy of an antibacterial agent (carbadox) in combination with added ZnO. The experiment was a 2 x 3 factorial arrangement in a randomized complete block design conducted in a total of 20 replicates. Carbadox was added at 0 or 55 mg/kg diet, and ZnO was added at 0, 1,500, or 3,000 mg Zn/ kg. A total of 918 pigs were weaned at an average 19.7 d of age. For the 28-d postweaning period, gains (P < 0.01), feed intakes (P < 0.05), and gain:feed ratios (P < 0.05) increased when dietary ZnO concentrations increased and when carbadox was added. These responses occurred in an additive manner. The results of these studies suggest that supplemental ZnO at 1,500 to 2,000 mg Zn/kg Zn improved postweaning pig performance, and its combination with an antibacterial agent resulted in additional performance improvements.     

Use of biochemical markers of osteoarthritis to investigate the potential disease-modifying effect of tibial plateau levelling osteotomy

OBJECTIVES: To evaluate the hypothesis that the concentration of the 1/20/5D4 epitope of keratan sulphate, cartilage oligomeric matrix protein and total sulphated glycosaminoglycans in synovial fluids from dogs with cranial cruciate ligament disease would be affected by tibial plateau levelling osteotomy. In addition, to evaluate the hypothesis that medial meniscal release or meniscal injury would alter the expression of these candidate biomarkers. METHODS: Forty-one dogs with naturally occurring cranial cruciate ligament disease were recruited prospectively. Synovial fluids were collected from the index joint before surgery and six weeks and six months postsurgery. Following tibial plateau levelling osteotomy, synovial fluids were assayed for 1/20/5D4 epitope of keratan sulphate and cartilage oligomeric matrix protein concentration using an inhibition ELISA and for sulphated glycosaminoglycans using a direct dye-binding assay. RESULTS: The sulphated glycosaminoglycans ratio did not change significantly during the study. Medial meniscal injury at entry was associated with lower concentrations of synovial fluid cartilage oligomeric matrix protein (P<0.05, unpaired t test). There was no association between medial meniscal release and the changes in marker concentrations, either from 0 to six weeks or 0 to six months. CLINICAL SIGNIFICANCE: Tibial plateau levelling osteotomy did not significantly alter the expression of the named candidate biomarkers. These findings reflect the limited nature of the arthrotomy or indicate that tibial plateau levelling osteotomy does not influence the progression of osteoarthritis (OA). From these studies, there is no evidence that tibial plateau levelling osteotomy affects cartilage metabolism.     

Stress-related behaviors among horses used in a therapeutic riding program

OBJECTIVE: To determine whether therapeutic riding resulted in higher levels of stress or frustration for horses than did recreational riding and whether therapeutic riding with at-risk individuals was more stressful for the horses than was therapeutic riding with individuals with physical or emotional handicaps. DESIGN: Observational study. ANIMALS: 14 horses in a therapeutic riding program. PROCEDURE: An ethogram of equine behaviors was created, and horses were observed while ridden by 5 groups of riders (recreational riders, physically handicapped riders, psychologically handicapped riders, at risk children, and special education children). Number of stress-related behaviors (ears pinned back, head raised, head turned, head tossed, head shaken, head down, and defecation) was compared among groups. RESULTS: No significant differences in mean number of stress-related behaviors were found when horses were ridden by recreational riders, physically handicapped riders, psychologically handicapped riders, or special education children. However, mean number of stress-related behaviors was significantly higher when horses were ridden by the at-risk children. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for horses in a therapeutic riding program, being ridden by physically or psychologically handicapped individuals is no more stressful for the horses than is being ridden in the same setting by recreational riders. However, at-risk children caused more stress to the horses, suggesting that the time horses are ridden by at-risk children should be limited both daily and weekly.