Reliable diagnostic and prognostic markers of sepsis are lacking, but essential in veterinary medicine. We aimed to assess the accuracy of C-Reactive Protein (CRP), protein carbonyls (PCO) and paraoxonase-1 (PON1) in differentiating dogs with sepsis from those with sterile inflammation and healthy ones, and predict the outcome in septic dogs. These analytes were retrospectively evaluated at admission in 92 dogs classified into healthy, septic and polytraumatized. Groups were compared using the Kruskal–Wallis test, followed by a Mann–Whitney U test to assess differences between survivors and non-survivors. Correlation between analytes was assessed using the Spearman’s test, and their discriminating power was assessed through a Receiver Operating Characteristic (ROC) curve. PON1 and CRP were, respectively, significantly lower and higher in dogs with sepsis compared with polytraumatized and clinically healthy dogs (p < 0.001 for both the analytes), and also in dogs with trauma compared with healthy dogs (p = 0.011 and p = 0.017, respectively). PCO were significantly increased in septic (p < 0.001) and polytraumatized (p < 0.005) as compared with healthy dogs. PON1 and CRP were, respectively, significantly lower and higher in dogs that died compared with survivors (p < 0.001 for both analytes). Ultimately, evaluation of CRP and PON1 at admission seems a reliable support to diagnose sepsis and predict outcomes.

In view of the enormous variability of dog breeds, breed-specific reference intervals (RIs) are recommended for use in veterinary clinical decision-making. The aim of this study was to determine whether RIs of the general canine population may be applied to the Italian Greyhound (Piccoli Levrieri Italiani or PLI), and to generate breed-specific RIs, where appropriate. Sixty-three privately owned clinically healthy fasted dogs were examined. Routine haematology and biochemistry were performed on 58 enrolled patients using the ADVIA 120 haematology analyzer and the Cobas Mira system, respectively. Changes in haematological and biochemical parameters depending on sex, age and attitude (resting vs. running dogs) were investigated. The results of PLI were compared with the RIs of the general canine population. In those cases in which these RIs were not validated, new RIs were generated according to the guidelines of the American Society of Veterinary Clinical Pathology. Pre-existing RIs were considered valid based on the recommendations by the Clinical & Laboratory Standards Institute (CLSI). RIs were higher for mean corpuscular haemoglobin (MCH), mean cell haemoglobin concentration (MCHC), cell haemoglobin concentration mean (CHCM) and lower for large unstained cells (LUC). A wider discrepancy between pre-existing and newly established RIs was found for some ADVIA parameters regarding red blood cell (RBC) or reticulocyte morphology. For total protein and cholesterol the new RIs were wider than the pre-existing ones, while albumin, calcium and iron were higher. This study suggests that most of the RIs published in veterinary textbooks cannot be validated for PLIs.

Oxidative stress (OS) is an imbalance between oxidants and anti-oxidants and plays an important rolein the aetiology and/or the progression of several diseases. Protein carbonyl (PCO) groups are so farused as biomarkers of OS in humans (Colombo et al, 2015).The aim of our study was to investigate whether PCOs are present in canine serum and if they can bemeasured spectrophotometrically using a method not yet validated in dogs.The presence of PCO was investigated by Western blotting after separation by SDS-PAGE of serum atdifferent dilutions (either before or after oxidation with 10% cigarette smoke extract). Protein labellingwith 2,4-Dinitrophenylhydrazine (DNPH, Brady's reagent) was followed by a two-steps incubation withprimary anti-dinitrophenyl-KLH antibodies (rabbit IgG fraction) and secondary goat anti-rabbit IgG, HRPconjugate. Signal was developed with Enhanced Chemiolumionescence (ECL).Serum PCO were quantified using a commercially available assay (Protein Carbonyl Content Assay Kit -Abcam, UK), based on derivatization of proteins with 1,4-dinitrophenylhydrazine (DNPH) andsubsequent formation of protein-conjugated dinitrophenylhydrazones (DNPs) with a peak absorbanceat 366 nm.Western blot showed an evident band of apparent MW of 69 kDa, consistent with carbonylated dogserum albumin. The spectrophotometric assay failed to demonstrate any signal at 366 nm using themanufacturer’s instructions or modified protocols.This study demonstrated that PCO are present in oxidized canine serum. However, thespectrophotometric assay employed in this study is not enough sensitive to detect PCOs. Furtherstudies are needed to assess whether this depends on the poor re-solubilisation of DNPs, or on the lowconcentration of PCOs in dogs.