Proliferative renal myxosporidiosis in spawning coho salmon (Oncorhynchus kisutch) in British Columbia and Washington

An unidentified myxosporean parasite (CKX) is described from the kidney of approximately 80% of spawning coho salmon Oncorhynchus kisutch (Walbaum) in British Columbia, Canada and Washington, United States of America. Morphological features were described using light and electron microscopy. Sequencing of polymerase chain reaction (PCR) amplified 18S ribosomal RNA gene and in situ hybridisation were used to further characterise CKX. The parasite occurred with a focal distribution within tubule epithelial cells, the tubule lumen and the interstitium as primary cells containing from one to at least 16 secondary cells. Luminal stages were degenerate and sporogony was not observed. In situ hybridisation using a digoxygenin-labelled DNA probe confirmed CKX to be the source of DNA used in PCR analyses. CKX 18S rDNA sequences were most similar (97%) to those of Sphaerospora oncorhynchi. Phylogenetic analysis revealed similarities among the 18S rDNA sequences of CKX, S. oncorhynchi and Myxidium lieberkuehni. CKX is hypothesised to be the abortive extrasporogonic development of a Sphaerospora sp. or Myxidium sp. occurring in immune-incompetent spawning and senescent salmon.     

Owners' perceptions of the health and performance of Pony Club horses in Australia

Pony Club is one of the leading junior equestrian organisations in the world, and was established to teach young people the many aspects of horsemanship. The aim of this study was to gain an understanding of the health and performance of Pony Club horses from the perspective of their owners. In-depth interviews were conducted with Pony Club members at two sites in Australia, and topics relating to participants' background with horses, horse attributes valued by participants, horse health and performance, and Pony Club-related matters. The in-depth interviews were taped, transcribed and analysed, by describing the themes and issues recorded in the dialogue and conducting cross-case analysis (finding similarities and dissimilarities between participants with respect to each of the above-mentioned topics). A total of 32 interviews were conducted. The participants' background with horses varied greatly. The horse attributes valued by > or =59% of participants included temperament, size, ability and suitability for riders. A range of issues relating to health and performance were important to > or =53% of participants, including horse temperament, nutrition, internal parasites, lameness and foot-care, and colic. Soundness and preventive health measures were rarely mentioned (< or =16% of participants). Friends or knowledgeable horse people were identified as the first point of contact for horse-health matters, and veterinarians were only used as a last resort or for serious problems. Members of Pony Club learned about their horses by trial and error. Optimal performance was described as a horse that was willing to do as the rider asked. Poor performance was usually the result of the horse misbehaving, and could include a resistance to rider commands, pigrooting and bucking.     

Plasma endotoxin concentration in horses: a methods study

Background: Accurate determination of plasma endotoxin concentration is critical for ex vivo and in vitro cellular and molecular studies of endotoxemia in horses. However, reports are conflicting with respect to anticoagulant, handling, and sample preparation.

Objective:

Objective: The purpose of this study was to determine the effect of blood sample fraction and handling time on measurement of endotoxin concentration in horses.

Methods:

Methods: Whole blood, anticoagulated with 3.8% (0.12 M) sodium citrate (9:1), was collected from 5 healthy horses. Whole blood (WB), platelet-rich plasma (PRP), and platelet-poor plasma (PPP) were spiked with endotoxin (2 EU/mL). Endotoxin-spiked WB samples were centrifuged immediately to generate PRP for measurement. Endotoxin concentration was subsequently measured by Limulus amebocyte assay at 0, 15, 30, 45, and 60 minutes. Assays were performed in triplicate and results were analyzed using Student's t -test, with significance set at P < .05.

Results:

Results: Mean endotoxin concentrations in 2 EU/mL-spiked WB were significantly different from those in PPP at all time points tested. Recovery of endotoxin in PRP generated from WB was significantly diminished after just 15 minutes.

Conclusion:

Conclusion: PRP generated from WB is significantly more reliable than PPP in determining endotoxin concentration ex vivo. Measurement of endotoxin in PRP generated from WB was significantly diminished after 15 min, identifying a time frame within which to process blood samples for endotoxin analysis.     

Analysis of differential protein expression in Actinobacillus pleuropneumoniae by Surface Enhanced Laser Desorption Ionisation--ProteinChip (SELDI) technology

Actinobacillus pleuropneumoniae (APP) is the aetiological agent of porcine pleuropneumonia. An increased understanding of its molecular basis of pathogenicity and vaccine development will be facilitated by the availability of sequence data from a complete genome which, by analogy to other bacteria, is predicted to encode many proteins in the molecular mass range 3-20kDa. However, conventional techniques to study bacterial protein expression, such as SDS-PAGE and 2-dimensional electrophoresis, typically focus on the 15-200kDa range. In this study we have evaluated Surface Enhanced Laser Desorption Ionisation-ProteinChip (SELDI) technology for the analysis of protein expression, in particular those of <20kDa, of APP grown under different environmental conditions. Cytoplasmic/periplasmic and outer membrane protein fractions were obtained from the APP wildtype serotype 1 strain 4074 grown in Brain Heart Infusion (BHI) broth (+different concentrations of NAD), BHI containing pig serum or defined medium. Optimum conditions for SELDI profiles included a sample size of 1 microg and the use of sinapinic acid as the energy absorbing matrix. In the <20kDa range, the SELDI profiles obtained from wild-type bacteria grown in rich medium plus 33-66% pig serum were most similar to those grown in defined medium. The SELDI profiles of extracts of the wild-type and of an rpoE mutant were similar although there were clear differences. The results suggest that SELDI is a useful complementary approach to conventional proteomic analytical methods with APP, and presumably other bacterial pathogens, being particularly suited for analysis of proteins in the <20kDa mass range.     

Intraosseous gentamicin perfusion of the distal metacarpus in standing horses

OBJECTIVE: To report tissue gentamicin concentrations after intraosseous (IO) perfusion in standing horses. STUDY DESIGN: In vivo study. ANIMALS OR SAMPLE POPULATION: Twelve horses. METHODS: Sedated horses had a cannulated cortical bone screw inserted into the dorsolateral aspect of the treated metacarpus and a tourniquet applied proximally. Gentamicin (2.2 mg/kg) diluted in sterile saline solution (0.1 mL/kg) was infused through the screw. Two horses were euthanatized at each time interval: 0, 2, 6, 12, 24, and 36 hours. Synovial fluid and bone samples were collected distal to the screw from both forelimbs. Gentamicin concentrations were measured using fluorescence polarization immunoassay. RESULTS: The highest synovial fluid gentamicin concentrations were 385+/-273 microg/mL (mean+/-SD) in the metacarpophalangeal joint, 225+/-205 microg/mL in the proximal interphalangeal joint, 215+/-205 microg/mL in the distal interphalangeal joint, 382+/-195 microg/mL in the digital flexor tendon sheath, and 206+/-161 microg/mL in the navicular bursa. The highest bone concentrations of gentamicin were 55+/-30 microg/g in the distal metacarpus, 34+/-27 microg/g in the proximal, 16+/-15 microg/g in the middle, and 16+/-2.2 microg/g in the distal phalanges, and 27+/-17 microg/g in the proximal and 24+/-11 microg/g in the distal sesamoid bones. CONCLUSION: Standing IO perfusion of gentamicin resulted in local antibiotic concentrations in the synovial structures and bones of the distal aspect of the limb that exceed the reported minimum inhibitory concentration of pathogens commonly implicated in equine orthopedic infections. CLINICAL RELEVANCE: Standing IO perfusion of gentamicin in the distal aspect of the limb should be considered for treatment of orthopedic infections of this region in horses.     

A modified ventral fixation for surgical management of atlantoaxial subluxation in 19 dogs

OBJECTIVE: To describe a modified ventral stabilization technique for surgical management of atlantoaxial subluxation in dogs and to evaluate the outcome. STUDY DESIGN: Retrospective clinical study. SAMPLE POPULATION: Nineteen client-owned dogs. METHODS: Medical records of 19 dogs with a radiographic diagnosis of atlantoaxial subluxation surgically managed by a modified ventral fixation technique (cortical screws, Kirschner wires, polymethylmethacrylate) were reviewed. Data on pre- and post-operative neurologic status, surgical technique, and complications were retrieved. Follow-up evaluation was performed at approximately 1 month. Telephone interview of the owner was used for long-term assessment (median follow-up for 17 surviving dogs was 10.5 months). RESULTS: Adequate reduction and stabilization was achieved in all dogs based on radiographic assessment immediately after surgery. Improved neurologic outcome occurred in 16 dogs at 1 month and in 15 dogs at follow-up; 2 dogs died of post-operative complications within 24 hours of surgery. One dog was euthanatized at the owners' request because of recurrent neck pain associated with implant failure after 1 month. Two dogs required surgery to remove broken and migrated implants, but further stabilization was not necessary. CONCLUSIONS: Adequate stabilization and improved neurologic outcome was achieved in most dogs. However, on account of the small size of the study and the variable neurologic signs of the dogs on admission, the surgical technique described could not be compared to those previously reported. CLINICAL RELEVANCE: The surgical technique described is an effective means of surgical treatment for atlantoaxial subluxation.     

Current research and theories on the pathogenesis of acute laminitis in the horse

A large number of studies have been undertaken in recent years aimed at furthering our understanding of the complex mechanisms underlying the common and debilitating condition of acute laminitis in the horse. Many of these studies have either reinforced or cast doubt on previously held theories on the pathogenesis of this disease, while others have suggested new mechanisms which may play a key role in its development. This review seeks to put the current hypotheses into the context of this recent body of evidence. While a unifying theory may not yet seem to be achievable, this review demonstrates that most of the current theories are not mutually exclusive. Studies utilising in vitro and in vivo models of the disease, particularly addressing the areas of inflammation, haemodynamic disturbances and enzyme activation in the hoof, as well as the preceding events occurring in the hindgut, have helped to explain many clinical observations of the disease and may possibly lead to more effective therapies and means of prevention in the future.     

Architecture of a fungal fatty acid synthase at 5 A resolution

All steps of fatty acid synthesis in fungi are catalyzed by the fatty acid synthase, which forms a 2.6-megadalton alpha6beta6 complex. We have determined the molecular architecture of this multienzyme by fitting the structures of homologous enzymes that catalyze the individual steps of the reaction pathway into a 5 angstrom x-ray crystallographic electron density map. The huge assembly contains two separated reaction chambers, each equipped with three sets of active sites separated by distances up to approximately 130 angstroms, across which acyl carrier protein shuttles substrates during the reaction cycle. Regions of the electron density arising from well-defined structural features outside the catalytic domains separate the two reaction chambers and serve as a matrix in which domains carrying the various active sites are embedded. The structure rationalizes the compartmentalization of fatty acid synthesis, and the spatial arrangement of the active sites has specific implications for our understanding of the reaction cycle mechanism and of the architecture of multienzymes in general.     

Organics captured from comet 81P/Wild 2 by the Stardust spacecraft

Organics found in comet 81P/Wild 2 samples show a heterogeneous and unequilibrated distribution in abundance and composition. Some organics are similar, but not identical, to those in interplanetary dust particles and carbonaceous meteorites. A class of aromatic-poor organic material is also present. The organics are rich in oxygen and nitrogen compared with meteoritic organics. Aromatic compounds are present, but the samples tend to be relatively poorer in aromatics than are meteorites and interplanetary dust particles. The presence of deuterium and nitrogen-15 excesses suggest that some organics have an interstellar/protostellar heritage. Although the variable extent of modification of these materials by impact capture is not yet fully constrained, a diverse suite of organic compounds is present and identifiable within the returned samples.