Plasma concentrations of endotoxin and platelet activation in the developmental stage of oligofructose-induced laminitis.

The link between the fermentation of carbohydrate in the equine large intestine and the development of acute laminitis is poorly understood. Absorption of endotoxin (lipopolysaccharide; LPS) into the plasma has been observed in one experimental model of laminitis, but does not cause laminitis when administered alone. Thus, the potential role of endotoxin is unclear. Platelet activation has previously been demonstrated in the developmental stage of laminitis. Equine platelets are more sensitive than leukocytes to activation by endotoxin, and can be activated directly by LPS in the low pg/ml range, activating p38 MAP kinase and releasing serotonin (5-HT) and thromboxane. The objectives of this study were firstly to determine whether endotoxin and platelet activation could be measured in the plasma of horses in the developmental phase of laminitis induced with oligofructose. Secondly, the time course of events involving platelet activation and platelet-derived vasoactive mediator production was investigated. Laminitis was induced in six Standardbred horses by the administration of 10 g/kg bwt of oligofructose. Plasma samples were obtained every 4h, and platelet pellets were obtained by centrifugation. LPS was measured using a kinetic limulus amebocyte lysate assay, and platelet activation was assessed by Western blotting for the phosphorylated form of p38 MAP kinase. Plasma 5-HT was assayed by HPLC with electrochemical detection and thromboxane B(2) was measured by radioimmunoassay. Clinical signs of laminitis and histopathologic changes were observed in lamellar sections from five of the six horses. Onset of lameness was between 20 and 30 h after the administration of oligofructose. LPS increased above the limit of detection (0.6 pg/ml) to reach a peak of 2.4+/-1.0 pg/ml at 8 h. TNFalpha was also detectable in the plasma from 12 to 24 h. There was a time-dependent increase in platelet p38 MAPK phosphorylation, which peaked at approximately 12 h (3.8+/-1.3 fold increase); plasma 5-HT and thromboxane increased steadily after this time (2.9+/-0.6 and 11.3+/-5.0 fold increases, respectively). These data indicate that small quantities of endotoxin may move into the circulation from the large intestine after the sharp decrease in pH that occurs as a result of carbohydrate fermentation. Correlating these findings with in vitro studies suggests that LPS may primarily activate platelets, leading indirectly to the activation of leukocytes. Therefore, endotoxin may contribute in the initiation of the early inflammatory changes observed in experimental models of acute laminitis.

Retrospective analysis of local injection site adverse reactions associated with 230 allogenic administrations of bone marrow-derived mesenchymal stem cells in 164 horses.

BACKGROUND: Bone marrow-derived mesenchymal stem cells (BM-MSCs) are frequently used in the treatment of musculoskeletal injuries. Fully characterised cells that are readily available for use is optimum. Allogenic BM-MSCs can satisfy the need for rapid treatment, however, their safety has been questioned. OBJECTIVES: Objectives were to characterise BM-MSCs from an adult donor horse, in vitro, and to identify and describe adverse reactions that occurred following their injection into other horses. We hypothesised that BM-MSCs capable of proliferation, differentiation and lacking MHC II from one donor could be implanted into another individual without significant adverse reactions and the frequency of adverse reactions in clinical cases would be similar to that previously reported for autologous BM-MSCs. STUDY DESIGN: Retrospective clinical study. METHODS: BM-MSCs were proliferated and characterised from one donor and cryopreserved for clinical use. Medical records for horses injected with allogenic BM-MSCs from this donor at a single hospital were used. After routine lameness exam, lesions were identified using diagnostic ultrasound or MRI. Post injection reaction was defined as increased pain, swelling, or heat at or near injection site, or increased lameness. Treatments required for each reaction were noted. RESULTS: BM-MSCs proliferated and underwent differentiation. Cells were found to be negative for MHC-II (<2%) and were viable after cryopreservation and shipping. Ten of 230 (4.35%) injections were noted to be associated with an adverse reaction. Adverse reactions occurred in synovial structures (n = 3) and in soft tissues (n = 7). MAIN LIMITATIONS: This investigation could underestimate the number and severity of reactions. Mild reactions, such as synovitis, may have been missed. Also, anti-inflammatory drugs could overshadow mild reactions, making them less likely to be detected. CONCLUSIONS: Fully characterised allogenic BM-MSCs originating from a single donor horse can be administered to horses with soft tissue injuries with a low rate of adverse reaction. The Summary is available in Portuguese - see Supporting Information.

Integrative veterinary medical education and consensus guidelines for an integrative veterinary medicine curriculum within veterinary colleges.

Integrative veterinary medicine (IVM) describes the combination of complementary and alternative therapies with conventional care and is guided by the best available evidence. Veterinarians frequently encounter questions about complementary and alternative veterinary medicine (CAVM) in practice, and the general public has demonstrated increased interest in these areas for both human and animal health. Consequently, veterinary students should receive adequate exposure to the principles, theories, and current knowledge supporting or refuting such techniques. A proposed curriculum guideline would broadly introduce students to the objective evaluation of new veterinary treatments while increasing their preparation for responding to questions about IVM in clinical practice. Such a course should be evidence-based, unbiased, and unaffiliated with any particular CAVM advocacy or training group. All IVM courses require routine updating as new information becomes available. Controversies regarding IVM and CAVM must be addressed within the course and throughout the entire curriculum. Instructional honesty regarding the uncertainties in this emerging field is critical. Increased training of future veterinary professionals in IVM may produce an openness to new ideas that characterizes the scientific method and a willingness to pursue and incorporate evidence-based medicine in clinical practice with all therapies, including those presently regarded as integrative, complementary, or alternative.

Fertility aspects in yearling beef bulls grazing endophyte-infected tall fescue pastures.

During a 2-year study, yearling beef bulls were used to determine the effects of grazing on endophyte-infected tall fescue on endocrine profiles, semen quality and fertilisation potential. Bulls were allotted to graze tall fescue pastures infected with Neotyphodium coenophialum (E+; n = 20 per year) or Jesup/MaxQ (Pennington Seed, Atlanta, GA, USA; NTE; n = 10 per year). Bulls were grouped by scrotal circumference (SC), bodyweight (BW), breed composites and age to graze tall fescue pastures from mid-November until the end of June (within each year). Blood samples, BW, SC and rectal temperatures (RT) were collected every 14 days. Semen was collected from bulls every 60 days by electroejaculation and evaluated for motility and morphology. The developmental competence of oocytes fertilised in vitro with semen from respective treatments was determined. Bulls grazing E+ pastures had decreased BW gain (P < 0.01), increased overall RT (P < 0.01) and decreased prolactin (P < 0.01) compared with animals grazing NTE pastures. Neither percentage of normal sperm morphology nor motility differed between bulls grazed on the two pasture types. Semen from E+ bulls demonstrated decreased cleavage rates (P = 0.02) compared with semen from NTE bulls. However, development of cleaved embryos to the eight-cell and blastocyst stages did not differ between the two groups. In conclusion, semen from bulls grazing E+ tall fescue resulted in decreased cleavage rates in vitro, which may lower reproductive performance owing to reduced fertilisation ability.

Preliminary study of laryngeal sacculectomy in horses, using a neodymium:yttrium aluminum garnet laser technique.

In an attempt to ablate the laryngeal saccule as an alternative method of sacculectomy (conventially done through a laryngotomy incision for laryngeal hemiplegia) a neodymium:yttrium aluminum garnet laser was used transendoscopically in noncontact fashion in 6 horses. The procedure was easy, quick, and labor-saving, with few complications. Endoscopically, the lased sacculectomy site appeared healed at postsurgical day 42. On postsurgical day 42, microscopic examination revealed mucosal remnants under the granulation bed. Laser energy caused thermal damage to tissues adjacent to the lased saccule. Additional work must be done to refine the technique before it can be recommended for clinical applications.

Laminar microvascular flow, measured by means of laser Doppler flowmetry, during the prodromal stages of black walnut-induced laminitis in horses.

OBJECTIVE: To measure changes in laminar microvascular blood flow (LMBF) over time in healthy horses and horses in the prodromal stage of black walnut-induced laminitis and to determine the effects of glyceryl trinitrate application on LMBF in horses with acute laminitis. ANIMALS: 10 healthy adult horses. PROCEDURE: Laser Doppler flowmetry was used to measure LMBF Baseline measurements were obtained, horses were given deionized water via a nasogastric tube, and measurements were obtained hourly for 12 hours. Twenty-four hours later, baseline measurements were again obtained, and horses were given black walnut extract. Measurements were obtained hourly for 12 hours or until development of Obel grade-3 laminitis. At this time, 5 horses were treated with phenylbutazone, and the other 5 were treated with phenylbutazone and glyceryl trinitrate, and measurements were obtained hourly for an additional 12 hours. RESULTS: LMBF was significantly decreased 1 and 2 hours after administration of the black walnut extract but then returned to near-baseline values for the next 6 hours. Eight hours after extract administration, there was a second significant decrease in LMBF that persisted until the end of the study. Glyceryl trinitrate had no effect on LMBF. Clinical signs of laminitis developed 8 to 12 hours after extract administration. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that in horses with black walnut-induced laminitis, there is an early decrease in LMBF followed by reperfusion prior to onset of clinical signs. Treatment with glyceryl trinitrate after development of clinical signs of laminitis did not have a significant effect on LMBF.

Evaluation of laser Doppler flowmetry for measuring coronary band and laminar microcirculatory blood flow in clinically normal horses.

Once daily for 3 days, laser Doppler flowmetry was used in 5 healthy, nonsedated adult horses to evaluate coronary band and laminar microcirculatory blood flow (MBF) in both forelimbs. The coronary band had significantly (P < 0.05) higher MBF than did the laminae on the days evaluated. Significant variation in MBF was not found over the 3-day measurement period in any one site. Significant (P < 0.05) variation was found in coronary band MBF among horses. This variation was not observed in laminar MBF. On occlusion of the digital arteries at the level of the fetlock, marked decrease in coronary band and laminar MBF was observed. Twenty minutes after IV administration of acetylpromazine, marked increase in coronary band and laminar MBF was observed. The technique was easily performed in standing nonsedated horses, did not inflict discomfort, lacked complications, and measurements were repeatable. This technique provides an index of digital MBF, either intermittently or continuously, avoiding introduction of invasive variables associated with other techniques.

Evaluation of use of dimethyl sulfoxide for intra-articular lavage in clinically normal horses.

The antebrachiocarpal and tarsocrural joints of 10 adult horses were randomly assigned to 1 of 4 groups. Groups were formulated and were treated as follows: group 1, control (arthrocentesis only); group 2, buffered lactated Ringer solution; group 3, 10% dimethyl sulfoxide (DMSO; w/v) in lactated Ringer solution; and group 4, 30% DMSO (w/v) in lactated Ringer solution. Joints were lavaged once with the respective solution. Prior to lavage and on days 1, 4, and 8 after lavage, all horses were evaluated for lameness and joint effusion; synovial fluid total and differential WBC counts, synovial fluid total protein concentration, and mucin clot quality were determined. Horses were euthanatized on day 8, and joints were evaluated grossly, histologically, and histochemically. Significant difference was not observed in effect of lactated Ringer solution, 10% DMSO, and 30% DMSO on any measured variable. At 24 hours after treatment, significant (P less than 0.05) difference in synovial fluid WBC numbers and total protein concentration was detected between control and treated joints. Eighty percent of lavaged joints had effusion 24 hours after treatment, compared with 30% of control joints. Gross, histopathologic, or histochemical differences were not detected between treated and control joints. Results of the study indicate that buffered lactated Ringer, 10% DMSO, and 30% DMSO solutions induce similar inflammatory changes in articular structures and significantly greater inflammatory reaction than does arthrocentesis alone.

Evaluation of a dosing method for studying ergonovine effects in cattle.

We evaluated the feasibility of using miniosmotic pumps as a way to continuously treat cattle with a singular ergot alkaloid (ergonovine) of known content, thus mimicking the natural fescue toxicosis disease state, but allowing study of specific alkaloid effects. Dosing animals with increasing amounts of ergonovine via miniosomotic pumps, followed by daily acquisition of plasma samples for high-performance liquid chromatographic determination of the alkaloid, resulted in stepwise increases in plasma ergonovine concentration. However, despite the detectable blood concentration of ergonovine, calves did not have typical clinical signs of ergot alkaloid toxicosis. Similarly, serum prolactin concentration was unaffected by ergonovine in these cattle, implicating some other alkaloid of endophyte-infested fescue as causative of the usual prolactin-suppressive response. The results confirm use of this animal dosing method to study biological effects of singular purified alkaloids of known amount, without bioavailability concerns. Thus, this dosing method will facilitate studies to determine the harmful effects of individual alkaloids found in toxic tall fescue, and ultimately, to alleviate their costly effects in cattle, horses, and other species.

Ascorbic acid as suspected cause of oxalate nephrotoxicosis in a goat.

Oxalate nephrotoxicosis was determined, by renal biopsy, to be the cause of azotemia in a goat. The origin of the oxalate was determined to be a high concentration of ascorbic acid that had been administered parenterally to the goat. Ascorbic acid has been documented as a cause of oxalate nephrotoxicosis in human beings.

Solitary osteochondroma of the nasal bone in a horse.

A 3-year-old Appaloosa stallion with a 4 cm x 4 cm x 2.5 cm mass protruding from his nasal bone was evaluated. Radiographs revealed an osseous mass, with a radiopaque outer margin and several radiolucent areas within the body of the mass. The mass was surgically removed and evaluated histopathologically. The histopathological diagnosis was osteochondroma. This case represent the first reported occurrence of an osteochondroma arising from intramembraneous bone in the horse.

Suspected buttercup (Ranunculus bulbosus) toxicosis with secondary photosensitization in a Charolais heifer.

A presumptive diagnosis of buttercup toxicosis with photosensitization secondary to hepatotoxicity was made in an 18-mo-old Charolais heifer. The differential diagnosis included salmonellosis, aflatoxicosis, bovine virus diarrhea, internal parasite infestation, and plant toxicosis with either primary or secondary photosensitization. All these possibilities were excluded except buttercup toxicosis with photosensitization secondary to hepatotoxicity. While this diagnosis was not absolutely confirmed, it was the most likely cause of the disease and raised the intriguing possibility that protoanemonin, buttercup's toxic principle, is hepatotoxic.