Myoplasmic calcium regulation in myotubes from horses with recurrent exertional rhabdomyolysis

OBJECTIVE: To determine whether alterations in myoplasmic calcium regulation can be identified in muscle cell cultures (myotubes) and intact muscle fiber bundles derived from Thoroughbreds affected with recurrent exertional rhabdomyolysis (RER). ANIMALS: 6 related Thoroughbreds with RER and 8 clinically normal (control) Thoroughbred or crossbred horses. PROCEDURES: Myotube cell cultures were grown from satellite cells obtained from muscle biopsy specimens of RER-affected and control horses. Fura-2 fluorescence was used to measure resting myoplasmic calcium concentration as well as caffeine- and 4-chloro-m-cresol (4-CMC)-induced increases in myoplasmic calcium. In addition, intact intercostal muscle fiber bundles were prepared from both types of horses, and their sensitivities to caffeine- and 4-CMC-induced contractures were determined. RESULTS: Myotubes of RER-affected and control horses had identical resting myoplasmic calcium concentrations. Myotubes from RER-affected horses had significantly higher myoplasmic calcium concentrations than myotubes from control horses following the addition of > or = 2mM caffeine; however, there was no difference in their response to 4-CMC (> or = 1 mM). Caffeine contracture thresholds for RER and control intact muscle cell bundles (2 vs 10mM, respectively) were significantly different, but 4-CMC contracture thresholds of muscle bundles from RER-affected and control horses (500 microM) did not differ. CONCLUSIONS AND CLINICAL RELEVANCE: An increase in caffeine sensitivity of muscle cells derived from a family of related RER-affected horses was detected in vitro by use of cell culture with calcium imaging and by use of fiber bundle contractility techniques. An alteration in muscle cell calcium regulation is a primary factor in the cause of this heritable myopathy.     

Effect of tilmicosin on chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages

OBJECTIVE: To evaluate chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages (AM) exposed to tilmicosin. ANIMALS: 12 healthy calves and 12 healthy pigs. PROCEDURES: Lungs were obtained immediately after euthanasia; AM were collected by means of bronchoalveolar lavage and density gradient centrifugation. Chemotactic activity was evaluated by exposing AM to lipopolysaccharide or macrophage inhibitory peptide during incubation with tilmicosin. Phagocytic activity was evaluated by incubating AM with tilmicosin for 24 hours and then with tilmicosin-resistant Salmonella serotype Typhimurium. Bactericidal activity was evaluated by incubating AM with tilmicosin (0, 10, or 20 microg/ml for bovine AM; 0 or 10 microg/ml or 10 microg/ml but washed free of tilmicosin for porcine AM) and then with Mannheimia haemolytica (bovine AM) or with Actinobacillus pleuropneumoniae or Pasteurella multocida (porcine AM). RESULTS: Tilmicosin had no significant effects on chemotactic or phagocytic activities of bovine or porcine AM. The time-course of bactericidal activity was best described by polynomial equations. Time to cessation of bacterial growth and area under the time versus bacterial number curve were significantly affected by incubation of AM with tilmicosin. CONCLUSIONS AND CLINICAL RELEVANCE: Results show that bactericidal activity of bovine and porcine AM was enhanced by tilmicosin, but not in proportion to the reported ability of AM to concentrate tilmicosin intracellularly. With or without exposure to tilmicosin, the time-course of bactericidal activity of bovine AM against M haemolytica and of porcine AM against A pleuropneumoniae or P multocida was too complex to be reduced to a simple linear equation.     

Evaluation of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals

OBJECTIVE: To determine efficacy of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals. DESIGN: Randomized clinical trial. ANIMALS: 165 foals. PROCEDURE: Foals were randomly assigned to 1 of 2 groups (hyperimmune plasma or nontreated controls). Foals with failure of passive transfer (FPT) of immunity were treated with hyperimmune plasma and evaluated as a third group. Foals that received plasma were given 950 ml between 1 and 10 days of age and between 30 and 50 days of age. A tracheobronchial aspirate was obtained from foals with clinical signs of respiratory tract disease for bacteriologic culture. RESULTS: A significant difference in incidence of pneumonia caused by R equi in foals with adequate passive transfer was not detected between foals that received plasma (19.1%) and nontreated foals (30%). Of 13 foals without FPT that received plasma and developed pneumonia caused by R equi, 12 developed disease prior to administration of the second dose of hyperimmune plasma. Incidence of undifferentiated pneumonia of all causes was not different between groups. CONCLUSION AND CLINICAL RELEVANCE: Intravenous administration of the commercially available hyperimmune plasma was safe, and the product contained high concentrations of anti-R equi antibodies. However, within this limited foal population, the difference in incidence of pneumonia caused by R equi observed between foals that received plasma and control foals was not significant.     

Use of low doses of ketamine administered by constant rate infusion as an adjunct for postoperative analgesia in dogs

OBJECTIVE: To compare indicators of postoperative pain and behavior in dogs with and without a low-dose ketamine infusion added to usual perioperative management. DESIGN: Prospective, randomized, blinded clinical study. ANIMALS: 27 dogs undergoing forelimb amputation. PROCEDURE: Dogs were anesthetized with glycopyrrolate, morphine, propofol, and isoflurane. Thirteen dogs were treated with ketamine IV, as follows: 0.5 mg/kg (0.23 mg/lb) as a bolus before surgery, 10 microg/kg/min (4.5 microg/lb/min) during surgery, and 2 microg/kg/min (0.9 microg/lb/min) for 18 hours after surgery. Fourteen dogs received the same volume of saline (0.9% NaCl) solution. All dogs received an infusion of fentanyl (1 to 5 microg/kg/h [0.45 to 2.27 pg/lb/h]) for the first 18 hours after surgery. Dogs were evaluated for signs of pain before surgery, at the time of extubation, and 1, 2, 3, 4, 12, and 18 hours after extubation. Owners evaluated their dogs' appetite, activity, and wound soreness on postoperative days 2, 3, and 4. RESULTS: Dogs that received ketamine infusions had significantly lower pain scores 12 and 18 hours after surgery and were significantly more active on postoperative day 3 than dogs that received saline solution infusions. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that perioperative administration of low doses of ketamine to dogs may augment analgesia and comfort in the postoperative surgical period.     

Sensitivity of test strategies used in the Voluntary Johne's Disease Herd Status Program for detection of Mycobacterium paratuberculosis infection in dairy cattle herds

OBJECTIVE: To evaluate sensitivities at the herd level of test strategies used in the Voluntary Johne's Disease Herd Status Program (VJDHSP) and alternative test strategies for detecting dairy cattle herds infected with Mycobacterium paratuberculosis. DESIGN: Nonrandom cross-sectional study. SAMPLE POPULATION: 64 dairy herds from Pennsylvania, Minnesota, Colorado, Ohio, and Wisconsin. Fifty-six herds had at least 1 cow shedding M. paratuberculosis in feces; the other 8 herds were free from paratuberculosis. PROCEDURE: For all adult cows in each herd, serum samples were tested for antibodies to M. paratuberculosis with an ELISA, and fecal samples were submitted for bacterial culture for M. paratuberculosis. Sensitivities at the herd level (probability of detecting infected herd) of various testing strategies were then evaluated. RESULTS: Sensitivity at the herd level of the testing strategy used in level 1 of the VJDHSP (use of the ELISA to test samples from 30 cows followed by confirmatory bacterial culture of feces from cows with positive ELISA result) ranged from 33 to 84% for infected herds, depending on percentage of cows in the herd with positive bacterial culture results. If follow-up bacterial culture was not used to confirm positive ELISA results, sensitivity ranged from 70 to 93%, but probability of identifying uninfected herds as infected was 89%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the testing strategy used in the VJDHSP will fail to identify as infected most dairy herds with a low prevalence of paratuberculosis. A higher percentage of infected herds was detected if follow-up bacterial culture was not used, but this test strategy was associated with a high probability of misclassifying uninfected herds.     

Associations between age or sex and prevalence of gastric ulceration in Standardbred racehorses in training

OBJECTIVE: To determine associations between age, sex, or medical treatment and prevalence and severity of gastric ulceration in Standardbred racehorses in training. DESIGN: Cross-sectional study. ANIMALS: 224 Standardbred racehorses in training. PROCEDURE: Gastroscopy was performed on each horse, and mucosal ulceration was graded from 0 (normal mucosa, no lesions) to 3 (extensive, often coalescing, lesions with areas of deep ulceration). Associations between age, sex, or treatment and prevalence and severity of ulcers were evaluated. RESULTS: Prevalence of gastric ulceration was 87%. Although there was little association between age and prevalence of ulcers, there was an association between age and severity of ulcers. Most 2-year-old horses (57.7%) had an ulcer score of 0 or 1. In all other age groups, most (58% to 82.61%) of horses had an ulcer score of 2 or 3. Although overall prevalence of ulceration was comparable among sex groups, the relative risk for gastric ulceration increased with age in castrated males, whereas it decreased in females and sexually intact males. CONCLUSIONS AND CLINICAL RELEVANCE: Gastric ulceration is common in Standardbred horses in race training. Severity is higher in horses > or = 3 years of age than in 2-year-old horses. Relative risk for ulceration increases with age in castrated males.     

Seroprevalence of antibodies specific for gram-negative core antigens in chickens on the basis of an Escherichia coli J5 enzyme-linked immunosorbent assay

Antibodies directed toward gram-negative core antigens (GNCAs) have been demonstrated in many mammalian species but to date are unexamined in any avian species. An enzyme-linked immunosorbent assay with phenol-killed whole cell Escherichia coli J5 was used to assess the presence of serum antibodies directed toward GNCAs in chickens. The first experiment consisted of collecting blood samples from randomly selected hens at egg laying ranches in northern California. The ages ranged from several days of age to 77 wk of age. Birds were classified into age groups (hatchling [1 day-4 wk], pullet [4-18 wk], pullet cycle [18-60 wk], and postmolt [>60 wk]) and husbandry style for titer comparison. The geometric mean titer (GMT) for all adult hens regardless of age was 2147. The geometric mean titers were 220, 5691, 2304, and 1776 for hatchlings, pullets, pullet cycle hens, and postmolt hens, respectively. The age group titer trends were similar to those of humans rather than those of farm animals in that the highest titers occurred during "adolescence" (pullets) and titers decreased slightly with maturity. The GMTs were 2870 for hens housed intensively and 1872 for those housed extensively. The second experiment looked at the progression of GNCA titers within individual birds over a 1-yr period. Individual titers increased slightly throughout the study time of the second experiment.     

Identification and distribution of a novel Malassezia species yeast on normal equine skin

This study aimed to investigate the distribution of Malassezia species yeasts on the skin of healthy horses. Acetate tape samples were obtained from the lip, axilla, interbulbar region, groin and anus of 12 healthy horses. The samples were stained and examined microscopically and sites harbouring yeast-like organisms were identified. Contact plates were applied to the skin at these sites and cultured at 26 degrees C and 32 degrees C. No growth was obtained on horse blood, Sabouraud's dextrose or modified Dixon's agar. A pure growth of a Malassezia-type organism was obtained on Sabouraud's dextrose agar enriched with oleic acid when it was incubated at 30 degrees C. It was identified by 26S ribosomal DNA D1/D2 sequence analysis as a member of the genus Malassezia, and most closely related to Malassezia sympodialis. However, the level of sequence divergence indicated that it was a novel species.     

The effect of a porcine-derived small intestinal submucosa product on wounds with exposed bone in dogs

OBJECTIVE: To determine the effect of a porcine-derived small intestinal submucosa product (PSIS) on healing time, epithelialization, angiogenesis, contraction, and inflammation of wounds with exposed bone on the distal aspect of the limbs of dogs. STUDY DESIGN: Prospective, controlled, experimental study. ANIMAL POPULATION: 10 young adult, purpose-bred, male Beagles. METHODS: Small wounds with exposed bone were created on the lateral aspect of metatarsal V and the medial aspect of metatarsal II on both hindlimbs. Three sheets of PSIS were sutured into the wounds of the treated limb, and the other limb served as a control. On day 10, punch biopsies of the medial metatarsal wounds were collected and were evaluated microscopically after routine hematoxylin and eosin and phosphotungstic acid hematoxylin (PTAH) staining. The lateral metatarsal wounds were evaluated by planimetry and laser Doppler perfusion imaging on days 7, 14, and 21. Time until complete wound healing was also recorded. The level of significance was set at P < or =.05 for all statistical analyses. RESULTS: Laser Doppler perfusion measurements were significantly higher in control wounds on day 7, but no differences were noted on days 14 and 21. No significant differences in planimetric values, histopathologic appearance, or time until complete wound healing were noted among treated and control groups. CONCLUSIONS: No objective differences in healing were noted between control wounds and wounds treated with PSIS. CLINICAL RELEVANCE: There appears to be no contraindication to the use of PSIS on clean wounds with exposed bone on the distal limbs of dogs. However, our objective data provides no evidence that this product affects epithelialization, contraction, or time to complete healing in wounds with exposed bone.