Anna Pasquini

Analytical performances of d-ROMs test and BAP test in canine plasma. Definition of the normal range in healthy Labrador dogs.

An high level of ROS (Reactive Oxygen Species), due to an increased production of oxidant species and/or a decreased efficacy of antioxidant system, can lead to oxidative stress, an emerging health risk factor involved in the aging and in many diseases, including inflammatory, infectious and degenerative disorders, either in humans or in animals. In the last years some assays panels have been developed to globally evaluate the oxidative balance by means of the concomitant assessment of ROS production and antioxidant system capability. In this report, the validation trials of d-ROMs (Reactive Oxygen Metabolites— derived compounds) and BAP (Biological Antioxidant Potential) tests in canine specie are described and also the specific referral ranges are calculated in a Labrador population. The results of linearity, precision and accuracy trials show that both tests exhibit good to excellent analytical performances. The possibility of measuring oxidative stress in vivo with simple, cheap and accurate tests, d-ROMs test and BAP test, provides for the veterinarians a very suitable tool to monitor oxidative stress and to correctly choice of eventual antioxidant supplementations in diseases proven related to oxidative stress in animals and particularly in dogs. Further studies will be useful to confirm this possibility.

Plasma lipoprotein concentrations in the dog: the effects of gender, age, breed and diet

Earlier studies of canine lipoprotein metabolism have frequently not taken into account such variables as age, gender, lifestyle or feeding status. In the last years, many changes to lifestyle and feeding of dogs have occurred. In this study, C-tot, C-HDL, C-LDL, triglycerides and lipoprotein fractions were determined in 251 healthy dogs by means of enzymatic methods and through the electrophoretic technique. All data were analysed by multifactor anova test to determine which factors (age, gender, breed and diet) have a statistically significant effect (p < 0.05) on the determined parameter and subsequently Bonferronis test was applied where necessary. Gender, age, breed and diet can significantly affect lipid metabolism, in particular lipoproteins involved in cholesterol plasma transport; on the contrary, triglycerides are not influenced by the same factors. The most important observation about age is the high level of C-LDL in puppies under 1 year of age. The highest cholesterol concentrations are found in Rottweiler but high values of plasma cholesterol are found also in Pyrenees Mountain dog and a great level of C-LDL in Labrador. Diet has shown a great influence on lipidic metabolism: dogs fed with different high-quality dry foods had significant differences in plasma cholesterol values (C-tot, C-HDL, C-LDL,), in particular, dogs fed with a diet rich in fish and fish-by-products have shown the lowest levels of C-tot, C-HDL and C-LDL.

Evaluation of oxidative stress in hunting dogs during exercise.

Exercise has been shown to increase the production of reactive oxygen species (ROS) to a point that can exceed antioxidant defenses, to cause oxidative stress. The aim of our trials was to evaluate oxidative stress and recovery times in trained dogs during two different hunting exercises, with reactive oxygen metabolites-derivatives (d-ROMs) and biological antioxidant potential (BAP) tests. A group of nine privately owned Italian hounds were included. A 20-min aerobic exercise and a 4-h aerobic exercise, after 30 days of rest, were performed by the dogs. Our results show an oxidative stress after exercise due to both the high concentration of oxidants (d-ROMs) and the low level of antioxidant power (BAP). Besides, the recovery time is faster after the 4-h aerobic exercise than the 20-min aerobic exercise. Oxidative stress monitoring during dogs exercise could become an interesting aid to establish ideal adaptation to training.

Redox status evaluation in dogs affected by mast cell tumour.

Oxidative stress status has been evaluated in depth in human medicine and its role in carcinogenesis has been clearly established. The purpose of this prospective study was to evaluate antioxidant concentrations and oxidative stress in dogs with mast cell tumours (MCTs) that had received no previous treatments, and to compare them to healthy controls. In 23 dogs with mast cell tumour and 10 healthy controls, oxidative status was assessed using the Reactive Oxygen Metabolites-derived compounds (d-ROMs) test, antioxidant activity was measured by the Biological Antioxidant Potential (BAP) test, and α-tocopherol levels were evaluated using high-performance liquid chromatography and ultraviolet analysis. At baseline, dogs with MCT had significantly higher d-ROMs (P < 0.00001) and lower BAP (P < 0.0002) compared with healthy controls. However, no significant difference was observed for α-tocopherol (P = 0.95). Results suggest that oxidative stress pattern and oxidative defence barrier are altered in dogs with newly diagnosed MCT compared with control dogs. Future studies are needed in order to assess the prognostic role of oxidative stress and to evaluate the impact of different therapeutic approaches.

Haematological Alterations Observed in Equine Routine Complete Blood Counts. A Retrospective Investigation

Abstract: Results on more than 1000 complete blood counts (CBC) performed on hot-blooded horses, including Thoroughbred, Standardbred, and Arabian, have been reviewed. All the data were processed and stored at the Laboratory of Haematology, Department Veterinary Clinic, University of Pisa, over the last 2 years, and involved samples withdrawn from both inpatients and outpatients of the clinic. The CBC is routinely requested as a part of a complete clinical evaluation of horses as well as during the course of disease or as a routine check during the training process. The CBC, using K3-EDTA blood samples, included a three cell population count performed on an electronic impedance cell counter (Genius, SEAC, Calenzano, Fi, Italy), and the microscopic evaluation of a stained blood smear. In addition, the refractometric assessment of total plasma proteins and fibrinogen (by the heat precipitation method) was also performed using EDTA plasma. The incidence of haematological changes in red blood cells (RBCs), subtyped as polycythaemia and anaemia, were 16.2%. Anaemia totalled 15.1%, including mild (packed cell volume (PCV) 31–26%) at 10.2%, moderate (PCV 25–17%) at 4.1%, and severe (PCV <16%) at 0.8%. The incidence of white blood cell (WBC) changes, subtyped as leucocytosis and leucopenia, was 38.4%. Leucocytosis totalled 30.5%, including mild (WBC 10–15 000/ml) at 22.5%, moderate (WBC 15–20 000/ml) at 7.7%, and severe (WBC >20 000/ml) at 0.3%. The incidence of leucopenia was 7.9% (WBC <5500/ml). Interpretation of the full leucogram (including variation on several leucocyte types and fibrinogen values) gave an incidence of physiological leucocytosis at 9.9%, stress leucocytosis at 5.3%, and inflammatory leucocytosis at 37.3%. Thrombocytopenia was frequently observed at 7.2%, whereas thrombocytosis was observed in only 0.3% of cases. Horse CBCs display less characteristic blood pictures in comparison to other species due to their particular haemopoietic system, such as the long half-life of RBCs, the occurrence of a non-sinusoid spleen, the fate and distribution of WBCs, and the rather high clumping phenomena of horse platelets in EDTA glass vials.

Canine Erythrocyte Morphology: Observations of a New Pattern, the “Quatrefoil” Erythrocyte

The “quatrefoil” erythrocyte pattern may be observed during erythrocyte morphology evaluation in canine complete blood counts (CBCs). “Quatrefoil” red blood cells (RBCs) have been documented in blood smears using both optical and scanning electron microscopy. In our examination of 3,958 CBCs, we found a prevalence of 3.89 % “quatrefoil” RBCs. These RBCs mostly occurred between the body and feathered edge of the blood smears. This pattern was not statistically related to the breed or sex, but to the age of the dog (older dogs; p < 0.0001). “Quatrefoil” RBCs are not significantly associated with other morphological erythrocyte changes. However, compared with populations of dogs without “quatrefoil” RBCs, dogs with this RBC pattern are significantly more likely to exhibit lower total leukocyte and neutrophil counts. Hypotheses describing the origin of “quatrefoil” RBCs include artifact effects, dacryocyte overlapping, and the effect of adhesion forces between two circulating RBCs in the blood stream (i.e., tank-trading and/or tumbling).

Corneal crystalline stromal dystrophy and lipidic metabolism in the dog.

Corneal crystalline stromal dystrophy and lipidic metabolism in the dog G. Barsotti & A. Pasquini & L. Busillo & M. Senese & G. Cardini & G. Guidi Published online: 7 August 2008 # Springer Science + Business Media B.V. 2008

Retrospective and Prospective Investigations about “Quatrefoil” Erythrocytes in Canine Blood Smears

The presence of unusual two RBCs patterns (so-called “quatrefoil RBCs,” qRBCs) on canine blood smears at Optical Microscope (OM) was seen during routine evaluation of CBCs. Two consecutive retrospective investigations were arranged including about 7,000 CBCs and clinical records and laboratory data from dogs showing qRBCs. Few samples with qRBCs were prepared for Scanning Electron Microscope (SEM). qRBCs were found in 6.89% (139 of 2016) and 8.47% (133 of 1569) of dogs and in 3.89% (154 of 3,958) and 4.47% (138 of 3,081) of CBCs (some dogs were tested more than once). Statistical analysis was significant for age groups (Chi squared, P < 0.0001), decreased total leukocyte and neutrophil counts (ANOVA, P < 0.0001), RBCs anisocytosis, polychromasia, and Howell-Jolly bodies (ANOVA, P < 0.018, <0.005, and <0.003, respectively). qRBCs were distributed in the area of feathered edge and at the smear side of body-feathered edge area in blood films. SEM ruled out the possibility of an optical illusion or an accidental overlap. qRBCs are associated with ageing of dogs, total leukocyte and neutrophil counts, and RBC anisocytosis, polychromasia, and Howell-Jolly bodies. Few hypotheses were discussed to explain the origin and meaning of this RBC arrangement.

Investigations about canine erythrocyte morphology: observation of a new modification, the ‘quatrefoil’ erythrocytes

LXV Meeting of the Italian Society for Veterinary Sciences (SISVet), Tropea-Drapia 2011—selected papers Full papers will be available in Pugliese A, Ferlazzo A, Gaiti A, Boiti C (2013) Trends in Veterinary Sciences—Current Aspects in Veterinary Morphophysiology, Biochemistry, Animal Production, Food Hygiene and Clinical Sciences. Springer-Verlag, Berlin Heidelberg Antonio Pugliese & Adriana Ferlazzo & Alberto Gaiti & Cristiano Boiti BIOLOGY & REPRODUCTION Seasonal effect on hematological and innate immune parameters in Sea bass Dicentrarchus labrax Pascoli F*, Negrato E, Poltronieri C, Radaelli G, Bertotto D Dipartimento di Scienze Sperimentali Veterinarie, Facolta di Medicina Veterinaria, Universita degli Studi di Padova. *Correspondence: francesco.pascoli@unipd.it