High‐mobility group box 1 as a surrogate prognostic marker in dogs with systemic inflammatory response syndrome

Objective – To evaluate various surrogate markers associated with the inflammatory and counter‐inflammatory responses with respect to mortality in dogs with systemic inflammatory response syndrome (SIRS). Design – Prospective observational study. Setting – Veterinary Teaching Hospital. Animals – Twenty‐eight dogs with naturally occurring diseases and SIRS from January 2007 to May 2009. Interventions – Upon admission to the veterinary hospital, history and baseline data from the physical examination, including parameters previously defined for meeting SIRS criteria, were documented. Heparinized blood samples were collected and plasma cytokines interleukin‐6 (IL‐6), IL‐10, and high‐mobility group box 1 (HMGB1) were measured by sandwich ELISA. Measurements and Main Results – In nonsurvivors, median plasma HMGB1 concentrations (0.718 μg/L, interquartile range [IQR]; 0.300–1.626 μg/L) and the ratio of HMGB1 to IL‐10 (2.236, IQR; 0.972–5.367) were significantly increased as compared with those found in survivors (0.300 μg/L, IQR; 0.300–0.312 μg/L for HMGB1; 1.017, IQR; 0.862–1.126 for the ratio of HMGB1 to IL‐10, P=0.007 and 0.024, respectively). Plasma IL‐6, IL‐10, and the ratio of IL‐6 to IL‐10 were not significantly different between groups. Among the parameters studied, HMGB1 and the ratio of HMGB1 to IL‐10 performed the best in discriminating outcome in dogs with SIRS according to receiver operator characteristic curve analysis. Conclusions – Increases in plasma HMGB1 concentration and the ratio of HMGB1 to IL‐10 may predict poorer outcomes in dogs with SIRS. The approach described may lead to reliable prognostic biomarkers and new therapeutic concepts in the study of SIRS in dogs.     

Troponin assays in the assessment of the equine myocardium

In 2000, troponin assays were adopted as the test of choice for detection of myocardial injury in man. This decision was made after extensive testing and followed a 60 year search for a biomarker of myocardial damage with sufficient analytical sensitivity and specificity. This has led to proliferation of assays for use in human medicine, each requiring extensive testing and validation before it could be made available on the open market for human use. The search for ever‐more analytically sensitive assays and for a standard reference material continues. The adoption of troponin testing in veterinary medicine followed shortly after its development for use in man, providing a much‐needed means of detecting and monitoring myocardial damage in horses. However, application of these tests in veterinary medicine has exclusively involved use of assays designed for and clinically validated in human patients. There is no mandated requirement for test validation in veterinary medicine and, while many of these assays have been shown to be capable of detecting equine troponin, the wide diversity of available tests, lack of validation, absence of protocols for their use and lack of standardisation make their application problematic. The objective of this review article is to address this issue, offering guidance where data are available and encouraging caution where there are none. Ultimately, the overall goal of this review is to examine critically the use of troponin assays in the horse and to promote the accurate and appropriate interpretation of valid results.     

Discrepancies between ergosterol and the phospholipid fatty acid 18:2ω6,9 as biomarkers for fungi in boreal forest soils

Ergosterol and the phospholipid fatty acid (PLFA) 18:2ω6,9 are frequently used as fungal biomarkers in studies on soils, and in accordance with the ideal for biomarkers of microorganisms they are thought to turn over rapidly after cell death and lysis. These biomarkers should also show the same patterns and responses to perturbations of the studied system. Here, I report strong correlations, in natural boreal forests of contrasting fertility, between free ergosterol and PLFA 18:2ω6,9 (r=0.821, P=0.007, n=9). Surprisingly, ergosterol, but not PLFA 18:2ω6,9, appears non-responsive to both large-scale tree girdling, which interrupts tree belowground C allocation to ectomycorrhizal fungi, and to long-term N-loading, which may have negative effects on both mycorrhizal and saprotrophic fungi. These results, therefore, question the use of ergosterol to monitor effects of soil perturbations on fungi in the field, but do not put into question the use of the biomarker in natural forest ecosystems.     

Determination of microbial community structures of shrimp floc cultures by biomarkers and analysis of floc amino acid profiles

Simple, rapid and reliable methods are required to monitor the microbial community change in aquatic pond for better animal performance. Four floc (suspended organic matter) samples were collected from outdoor raceways and tanks used for culturing Pacific white shrimp Litopenaeus vannamei. Twenty‐two chlorophyll (Chl) and carotenoid pigments were separated, identified and quantified using high‐performance liquid chromatography–ultraviolet/Vis‐mass spectrometry in the freeze‐dried floc samples. Algal community composition (diatoms, chlorophytes, cyanobacteria, dinoflagellates and cryptophytes) was determined by measuring concentrations of the respective taxonomic biomarkers (carotenoid fucoxanthin, lutein, zeaxanthin, peridinin and alloxanthin) as independent variables and Chl a as the dependent variable using a multiple regression model. This analysis found that the phytoplankton community of the floc samples from two groups of shrimp tanks (32 g L^?1‐salinity) were diatom‐dominated (81.7% and 84.4%); and two floc samples from shrimp raceways (5 and 18 g L^?1‐salinity) were chlorophyte‐dominated (75.4% and 82.3%). Assessment of total algal and bacterial biomass by quantification of Chl a and muramic acid, respectively, indicated that the 18 g L^?1‐salinity raceway sample was bacteria‐dominated, whereas the other three floc samples were algae‐dominated. Sample protein quality was evaluated by its essential amino acid (AA) score and index. Arginine and lysine were found to be the two most limiting AAs for all floc samples.     

The effect of noncardiac disease on plasma brain natriuretic peptide concentration in dogs

Objective – To evaluate the effects of noncardiac disease on c‐terminal brain natriuretic peptide (cBNP) concentrations in dogs. Design – Prospective observational study. Setting – Urban university veterinary hospital. Animals – Thirty‐eight apparently healthy dogs, 28 dogs with cardiac disease (14 CHF, 14 non‐CHF), and 81 dogs with primary noncardiac diseases. Interventions – none. Materials and Methods – Plasma was collected from each dog and analyzed for active (cBNP) B‐type natriuretic peptide using an assay that is being investigated for commercial use (Biosite). Measurements and Main Results – Dogs with CHF had significantly higher plasma cBNP concentrations than dogs with subclinical cardiac disease, apparently healthy dogs, or dogs with primary noncardiac disease. However, 21% (28/133) of dogs without CHF (including healthy dogs, dogs with primary noncardiac disease, and dogs with subclinical cardiac disease) had cBNP concentrations above previously identified diagnostic thresholds for CHF, reiterating the importance of reestablishing new diagnostic cutoffs when considering comorbidities affecting B‐type natriuretic peptide levels. Conclusions – A clinically relevant proportion of nondyspneic dogs with primary noncardiac diseases have increased cBNP concentrations that exceed previously identified diagnostic thresholds, potentially limiting the ability of this test to identify CHF when noncardiac comorbidities exist. Interpretation of increased cBNP concentrations in such cases must be appropriately interpreted with further diagnostic investigation.     

Microbial communities, biomass, and activities in soils as affected by freeze thaw cycles

Two Finnish agricultural soils (peat soil and loamy sand) were exposed to four freeze-thaw cycles (FTC), with a temperature change from −17.3±0.4 °C to +4.1±0.4 °C. Control cores from both soils were kept at constant temperature (+6.6±2.0 °C) without FTCs. Soil N₂O and CO₂ emissions were monitored during soil thawing, and the effects of FTCs on soil microbes were studied. N₂O emissions were extremely low in peat soil, possibly due to low soil water content. Loamy sand had high N₂O emission, with the highest emission after the second FTC. Soil freeze-thaw increased anaerobic respiration in both soil types during the first 3-4 FTCs, and this increase was higher in the peat soil. The microbial community structure and biomass analysed with lipid biomarkers (phospholipid fatty acids, 3- and 2- hydroxy fatty acids) were not affected by freezing-thawing cycles, nor was soil microbial biomass carbon (MIB-C). Molecular analysis of the microbial community structure with temperature gradient gel electrophoresis (TGGE) also showed no changes due the FTCs. These results show that freezing and thawing of boreal soils does not have a strong effect on microbial biomass or community structure.     

Early diagnosis of acute kidney injury subsequent to severe hypotension and fluid resuscitation in anaesthetized dogs

ObjectivesTo document changes in urinary biomarker concentration and conventional diagnostic tests of acute kidney injury (AKI) following hypotension and fluid resuscitation in anaesthetized dogs.Study designExperimental, repeated measures, prospective study.AnimalsA group of six male adult Greyhound dogs.MethodsFollowing general anaesthesia, severe hypotension was induced by phlebotomy, maintaining mean arterial blood pressure (MAP) < 40 mmHg for 60 minutes, followed by resuscitation with intravenous gelatine solution to maintain MAP > 60 mmHg for 3 hours. Following euthanasia, renal tissue was examined by light microscopy (LM) and transmission electron microscopy (TEM). Urinary and serum concentrations of neutrophil gelatinase-associated lipocalin (NGAL), cystatin C (CysC), and gamma-glutamyl transpeptidase (GGT), serum creatinine and urine output were measured at baseline and hourly until euthanasia. Data are presented as mean and 95% confidence interval and analysed using repeated measures analysis of variance with Dunnett’s adjustment, p < 0.05.ResultsStructural damage to proximal renal tubular cells was evident on LM and TEM. Urinary biomarker concentrations were significantly elevated from baseline, peaking 2 hours after haemorrhage at 19.8 (15.1–25.9) ng mL^–1 NGAL (p = 0.002), 2.54 (1.64–3.43) mg mL^–1 CysC (p = 0.009) and 2043 (790–5458) U L^–1 GGT (p < 0.001). Serum creatinine remained within a breed-specific reference interval in all dogs. Urinary protein–creatinine ratio (UPC) was significantly elevated in all dogs from 1 hour following haemorrhage.Conclusions and clinical relevanceUrinary NGAL, CysC and GGT concentrations, and UPC were consistently elevated within 1 hour of severe hypotension, suggesting that proximal renal tubules are damaged in the earliest stage of ischaemia-reperfusion AKI. Measurement of urinary biomarkers may allow early diagnosis of AKI in anaesthetized dogs. Urinary GGT concentration and UPC are particularly useful as they can be measured on standard biochemistry analysers.     

前列腺素类及其受体在动物慢性炎症中的促炎及指示作用

动物慢性炎症是各种慢性疾病的基础,包括慢性乳腺炎、代谢综合征、关节炎、伤口感染、血管疾病和癌症等。前列腺素类(PGs)作为一种典型的急性炎症介质在近年来的动物试验研究中被证明在慢性炎症的过渡和维持中发挥着重要作用,主要包括:1)细胞因子信号传导扩增;2)促进获得性免疫并诱导长期免疫炎症;3)募集炎性细胞以在炎症部位交替活性细胞群;4)参与组织重塑。因此,PGs信号传导可能作为慢性炎症的治疗靶点。鉴于慢性炎症对动物健康的影响以及PGs在动物慢性炎症中的重要性,本文总结了PGs及其受体的分类以及PGs作为典型急性炎症介质的特点,重点描述了PGs在动物慢性炎症过程中发挥的促炎作用,并概述了前列腺素H2D-异构酶(PTGDS)与脂质运载蛋白型前列腺素D合成酶(L-PGDS)对奶牛慢性乳腺炎的指示作用。