Evaluation of point-of-care tests for diagnosis of disseminated intravascular coagulation in dogs admitted to an intensive care unit.

OBJECTIVE: To evaluate the accuracy of point-of-care tests for the diagnosis of disseminated intravascular coagulation (DIC) in dogs and assess the correlation and agreement of results between point-of-care and laboratory tests in the evaluation of hemostatic function. DESIGN: Prospective case series. ANIMALS: 59 critically ill dogs (affected dogs) with clinical signs of diseases known to predispose to DIC and 52 clinically normal dogs. PROCEDURES: Accuracy of the point-of-care tests (activated clotting time [ACT], estimated platelet count and number of schizocytes from a blood smear, plasma total solids [TS] concentration, and the protamine sulfate test) was evaluated, using receiver operating characteristic curves and likelihood ratios. A strategy, using likelihood ratios to calculate a posttest probability of DIC, was tested with 65% used as a threshold for initiation of treatment. Results of laboratory tests (coagulogram and plasma antithrombin III activity) were used as the standard for comparison in each dog. RESULTS: ACT and estimated platelet count provided the best accuracy for detection of DIC. The plasma TS concentration, schizocyte number, and protamine sulfate test had poor accuracy. The strategy using post-test probability of DIC identified 12 of 16 affected dogs that had DIC. Estimated platelet count was correlated and had acceptable clinical agreement with automated platelet count (r = 0.70). The plasma TS (r = 0.28) concentration and serum albumin (r = 0.63) concentration were not accurate predictors of plasma antithrombin III activity. The ACT did not correlate with activated partial thromboplastin time (r = 0.28). CONCLUSIONS AND CLINICAL RELEVANCE: Strategic use of likelihood ratios from point-of-care tests can assist clinicians in making treatment decisions for dogs suspected to have DIC when immediate laboratory support is unavailable.     

Isolation and partial characterization of rainbow trout (Oncorhynchus mykiss) gill mucin

Gill mucin from rainbow trout was isolated utilizing two rounds of cesium chloride density ultracentrifugation followed by gel filtration on Sepharose CL-2B. Neither density ultracentrifugation nor gel filtration alone was sufficient for purification of the mucin. Isolated gill mucin had a density of 1.5 g/ml and eluted at the void volume of the Sepharose CL-2B column. Silver-stained reducing 6% polyacrylamide gel electrophoresis of gill mucin produced a band at the origin with a smear entering the separating gel. There was no evidence of a link protein in gill mucin on reducing 12% polyacrylamide gel electrophoresis. Gill mucin had an amino acid profile similar to that of mucins in other species. Specifically, 35.1% of the total amino acids were represented by threonine and serine, while another 27.5% were alanine and proline. Gill mucin contained galactose (26.7 ± 3.2%), galactosamine (22.5 ± 4.4%), glucose (16.6 ± 8.7%), fucose (16.1 ± 1.5%), glucosamine (12.0 ± 1.9%) and mannose (5.1 ± 4.4%). Uronic acid levels from purified mucin were very low (0.7 ± 0.1%). Sialic acid was also present (0.06 g/g of mucin protein). The periodic acid-Schiff assay routinely utilized for detection of mucins was relatively insensitive for detection of gill mucin (6 × less sensitive than for pig gastric mucin) so a rabbit antiserum was raised. The antiserum produced profiles similar to the periodic acid-Schiff assay of fractions following gel filtration. Immunofluorescence of formalin-fixed rainbow trout gill tissue sections showed that the antiserum detected mucin within branchial goblet cells.     

Production and Germination of Conidia of Trichoderma stromaticum, a Mycoparasite of Crinipellis perniciosa on Cacao

ABSTRACT Growth characteristics of the fungus Trichoderma stromaticum, a mycoparasite on the mycelium and fruiting bodies of Crinipellis perniciosa, the causal agent of witches'-broom disease of cacao, were evaluated under controlled environmental conditions. The ability of T. stromaticum to produce conidia and germinate on dry brooms was evaluated at three constant temperatures (20, 25, and 30 degrees C) and two constant relative humidities (75 and 100%). T. stromaticum produced abundant conidia on brooms at 100% relative humidity and incubation temperatures of 20 and 25 degrees C, but none at 30 degrees C. Sporulation of T. stromaticum was not observed at 75% relative humidity at any temperature. At 100% relative humidity and either at 20 or 25 degrees C, treatment of brooms with T. stromaticum suppressed C. perniciosa within 7 days. In contrast, at 30 degrees C, treatment with T. stromaticum had no effect on the pathogen in brooms maintained at either 75 or 100% relative humidity. Mycelium of C. perniciosa grew from brooms at all temperatures at 100% relative humidity. Conidial germination on broom tissue approximated 80% at temperatures from 20 to 30 degrees C. Results suggest that applying T. stromaticum under high-moisture conditions when the air temperature is below 30 degrees C may enhance the establishment of this mycoparasite in cacao plantations.     

Epiglottic augmentation by use of polytetrafluoroethylene to correct dorsal displacement of the soft palate in a standardbred horse.

A 2-year-old 400-kg Standardbred colt with a history of exercise intolerance was determined to have dorsal displacement of the soft palate (DDSP). Standing endoscopy revealed a soft, flaccid, and hypoplastic epiglottis and arytenoid cartilages that moved symmetrically and abducted fully. Lateral laryngeal radiography was used to calculate a corrected thyroepiglottic length of 6.48 cm. Videoendoscopy during exercise on a high-speed treadmill at standardized treadmill speeds was performed weekly for 3 consecutive weeks. The soft palate dorsally displaced at each examination. A sternohyoideus/sternothyroideus myectomy was followed by 2 weeks of stall rest and 2 weeks of treadmill training. Because endoscopy during exercise at standardized treadmill speeds caused the horse to dorsally displace the soft palate once a week for 3 weeks after the myectomy, this surgery was deemed ineffective in this horse. Epiglottic augmentation was performed by injecting 7 ml of polytetrafluoroethylene into the ventral submucosal space of the epiglottis. After 21 days of stall rest and 5 weeks of pasture rest, the horse was placed on a 2-month treadmill conditioning program. Using endoscopy at standardized treadmill speeds, exercise efforts to encourage DDSP were unsuccessful, which suggested that epiglottic augmentation by use of polytetrafluoroethylene was an effective surgical treatment to correct dorsal displacement of the soft palate in this horse. It is believed that polytetrafluoroethylene increases the rigidity of the epiglottis, allowing it to maintain the ventral position of the soft palate.     

Hematology and biochemistry reference values for Ontario swine.

The purpose of this study was to establish blood reference values for Ontario swine from various age groups. Weanling pigs, feeder pigs, gilts and sows on 11 randomly selected swine farms were sampled using the orbital sinus bleeding technique. Routine hematological and biochemical determinations were performed using whole blood and serum. For the variables examined in each age group the means, the standard deviations and the 95% upper and lower limits were calculated.     

The Piromen Test as an Assay of Bone Marrow Granulocyte Reserves in the Calf I. Studies on Bone Marrow and Peripheral Blood Leukocytes

The use of Piromen, a polysaccharide complex of Pseudomonas aeroginosa, has been investigated in 21 calves by a test of marrow granulocyte reserves. The maximal increase in peripheral granulocytes (ΔG) was determined following multiple intravenous and sub-cutaneous injections of Piromen at various time intervals and was correlated with the mature marrow granulocytes on bone marrow smear differentials. Five ug Piromen per kg body weight, by the subcutaneous route, was found to give a mean ΔG of 5200/mm³, very similar to the ΔG of 5300/mm³ obtained in man with 0.1 ug/kg intravenous Piromen injections. Clinical effects in calves were minimal with the subcutaneous route as compared to the response following intravenous Piromen injections.     

Virulence of Flavobacterium psychrophilum isolates in rainbow trout Oncorhynchus mykiss (Walbaum)

Flavobacterium psychrophilum, the causative agent of bacterial cold‐water disease (BCWD) in freshwater‐reared salmonids, is also a common commensal organism of healthy fish. The virulence potential of F. psychrophilum isolates obtained from BCWD cases in Ontario between 1994 and 2009 was evaluated. In preliminary infection trials of rainbow trout juveniles, significant differences (0% to 63% mortality) in the virulence of the 22 isolates tested were noted following intraperitoneal injection with 10⁸ cfu/fish. A highly virulent strain, FPG 101, was selected for further study. When fish were injected intraperitoneally with a 10⁶, 10⁷ or 10⁸ cfu/fish of F. psychrophilum FPG 101, the 10⁸ cfu/fish dose produced significantly greater mortality (p < 0.05). The bacterial load in spleen samples collected from fish every 3 days after infection was determined using rpoC quantitative polymerase chain reaction amplification and by plate counting. Bacterial culture and rpoC qPCR were highly correlated (R² = 0.92); however, culture was more sensitive than the qPCR assay for the detection of F. psychrophilum in spleen tissue. Ninety‐seven per cent of the asymptomatic and the morbid fish had splenic bacterial loads of <2.8 log₁₀ gene/copies and >3.0 log₁₀ gene copies/reaction, respectively, following infection with 10⁸ cfu/fish.     

Function and heterogeneity of fish lectins

Lectins are primordial molecules with multiple known functions. They have been known to exist in fish and other animals for decades and were initially identified as (hem)agglutinins. Demonstration of the importance of vertebrate lectins in innate immunity is a recent effort and is still largely unrealised for fish. This mini-review will tabulate those fish lectins identified since the last major review. In addition, particular lectins for which either functional relevance or functional or structural heterogeneity has been demonstrated are discussed in greater detail.