Changes in the salivary metabolome in canine hypothyroidism: A pilot study.

Hypothyroidism is the most commonly diagnosed endocrine disorder in dogs. It produces a deficiency of thyroid hormones which impacts negatively the dog's quality of life. The objective of this study is to evaluate the possible changes in the salivary metabolic profile in dogs with hypothyroidism. For this purpose, targeted metabolomics analysis performed by LC/MS analysis was made in saliva samples from a group of dogs with hypothyroidism and a group of healthy dogs. Twenty-three metabolites showed a significant decrease between hypothyroid and healthy dogs, most of these associated with thyroid hormone synthesis, catecholamine synthesis, and tyrosine and phenylalanine metabolism. Based on the results, it can be stated that hypothyroidism produces changes in the metabolome of saliva and some of them can reflect the metabolic changes presented in the disease and could serve as a potential biomarker of this condition.

Evaluation of Changes in Metabolites of Saliva in Canine Obesity Using a Targeted Metabolomic Approach.

Obesity is a common problem in pet dogs, affecting half of the general population in some countries. Excess body weight causes several disorders and has a negative impact on dogs' quality of life. The use of metabolomics allows the identification of metabolite traces from the metabolic pathways involved in pathological processes. This study aimed to evaluate salivary metabolite variations in dogs with obesity. The salivary samples of 19 dogs were analyzed using a targeted metabolomic approach, through which 234 metabolites were quantified. Of these, multivariate analysis identified 27 different metabolites altered in dogs with obesity compared with control dogs. These metabolites were mainly classified as amino acids, glycerides, sphingolipids, glycerophospholipids, and acylcarnitines. Some of the changes in these metabolites reflect the insulin resistance status related to obesity in dogs. Overall, it can be concluded that the salivary metabolome of obese dogs reflects the metabolic changes occurring in obesity and could be a source of potential biomarkers for this complex condition.

Metabolic profiling of serum from dogs with pituitary-dependent hyperadrenocorticism.

Hyperadrenocorticism (HAC) is one of the most common endocrine diseases in dogs characterized by excessive cortisol production caused by an adrenocorticotropic hormone (ACTH)-secreting tumor, namely pituitary-dependent HAC (PDH) or cortisol-secreting adrenal tumor. Metabolomics presents the ability to identify small molecule metabolites. Thus, the use of metabolomics techniques in canine PDH can provide information about the pathophysiology and metabolic changes in this disease. This study aimed to identify and compare differences in serum metabolites between dogs with PDH and healthy dogs. The metabolomic profile of 20 dogs diagnosed with PDH was compared with 20 healthy dogs using liquid chromatography/mass spectrometry (LC/MS), and metabolite discrimination was performed using partial least squares-discriminant analysis (PLS-DA), the variable important in projection (VIP) and fold changes (FC) group-wise comparisons. The hypergeometric test identified the significantly altered pathways. A total of 21 metabolites were found to be significantly different between the two groups. The major alterations were found in arachidonic and decanoic acid, and phospholipids related to phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI). These metabolites are related to insulin resistance and other complications (i.e. hypertension). Our results indicate that PDH produces changes in serum metabolites of dogs, and the knowledge of these changes can aid to better understanding of pathophysiological processes involved and contribute to potentially detect new biomarkers for this disease.

MCP-1, KC-like and IL-8 as critical mediators of pathogenesis caused by Babesia canis.

Canine babesiosis caused by the intraerythrocytic protozoan parasite Babesia canis is a tick-borne disease characterized by a host response that involves both cellular and humoral immunity. This study focuses on the secretion of cytokines Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), Keratinocyte Chemotactic-like (KC-like), Interleukins (IL)-2, IL-7, IL-8, IL-10, IL-15, IL-18 and Monocyte Chemotactic Protein-1 (MCP-1) in babesiosis caused by Babesia canis upon treatment with Imizol®. We assessed time dependent changes in cytokine levels and tested whether these changes correlate with pathogenesis of the disease. Sixteen healthy dogs and 31 dogs infected with Babesia canis, of which 18 showed complications, were treated with Imizol®. One dog died during the study (3.2%). Longitudinal study was perfomed by monitoring dogs at the first day of presentation (day 1) and 6 days later (day 7). Our results show that higher MCP-1 levels on day 1 are positively associated with the occurrence of complications, (complicated vs. uncomplicated; p = 0.00016). A similar pattern was observed for KC-like on day 1 (p = 0.0326) and day 7 (p = 0.044). Moreover, babesiosis caused by B. canis produced a steady increase in IL-8 levels with a moderate to strong negative correlation with erythrocyte counts and hematocrit in uncomplicated diseased dogs only (Spearman's rank correlation coefficient rs = -0.582 and rs = -0.598 respectively). Like for MCP-1, KC-like levels also differed in complicated and uncomplicated diseased dogs on day 1 (p = 0.03236) and day 7 (p = 0.044). Furthermore, KC-like levels were strongly correlated with IL-8 levels (rs = 0.663-0.7) and non-segmented neutrophil counts (rs = 0.572-0.732) in both diseased groups. Analysis of ROC suggests the use of serum levels of MCP-1 and IL-7 as predictors of the occurrence of complications with an AUC of 0.906 and 0.896 respectively and linear combinations of MCP-1, KC-Like, IL-7 and GM-CSF with values up to AUC = 0.983. Cytokine cluster analysis presented in this study can contribute to a better understanding of the pathogenesis of babesiosis and serve as a prognostic tool for the early detection of cases with highest likelihood of developing complications. Overall, our studies show that infection by B. canis elicits a cytokine pattern that is distinct from that observed with B. rossi, and that some of the inflammatory mediators can be useful to predict complications. Our results also suggest targets for the development of novel therapeutic strategies in babesiosis caused by B. canis.

Serum concentrations of eicosanoids and lipids in dogs naturally infected with Babesia canis.

Canine babesiosis is a tick-borne disease with world-wide significance caused by intraerythrocytic protozoa of the genus Babesia. The eicosanoids, as inflammatory mediators, are involved in the regulation of the immune response and inflammatory reaction. Metabolism of lipids is of great importance in babesiosis. In this study it was aimed to investigate the dynamics of serum concentration of prostaglandin E2 (PGE2), thromboxane B2 (TxB2), leukotriene B4 (LTB4), triglycerides, total cholesterol (Chol), HDL- and LDL-cholesterol in dogs naturally infected with Babesia canis and healthy dogs. Both groups were measured for all parameters on the admission day and on the first, second and seventh day of the disease. Dogs that were included in this study had systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS). It was demonstrated that the level of LTB4, PGE2, TxB2 in dogs naturally infected with B. canis significantly changed during the disease. The level of LTB4 was significantly higher during the study, while the concentration of PGE2 was significantly higher second, third and seventh day of disease in relation with healthy dogs. The level of TxB2 was significantly lower at the beginning of the disease, but after seven days concentration was significantly higher. Both group of patients with SIRS and MODS had significantly higher level of LTB4. Substained high concentrations of PGE2 were observed in dogs with MODS after therapy but not in dogs with SIRS, and LTB4 followed a similar tendency. On the other hand, increases in TxB2 were only significant in dogs with SIRS. The lipid profile in naturally infected dogs with B. canis infection was significantly changed. Further studies are needed to assess the prognostic values of lipid mediators in dogs with B. canis infection, and the ability of these markers to predict the progress of SIRS and MODS.

Molecular evidence of natural infection with Babesia canis canis in Croatia.

The aim of the present study was to detect and characterise the species and subspecies of Babesia spp. that cause canine babesiosis in Croatia. Twenty-eight dogs with typical signs of babesiosis (lethargy, anorexia, fever, dark urine and thrombocytopenia) were included in this study. Their blood smears showed the presence of Babesia canis . The results showed the detection of one subspecies, namely Babesia canis canis using PCR, and subsequent sequence analysis demonstrated portions of the nss rRNA gene in 27 out of 28 samples. Sequence analysis of the isolates showed 100% identity in 11 samples, 99.7% identity (one nucleotide difference) in 11 samples and 99.4% identity (two nucleotides difference) in 5 samples with B. canis canis . The results of this study confirm the presence of B. canis canis in infected dogs in Croatia and demonstrate a slightly new genetic variant of Babesia subspecies.

Alterations in some blood coagulation parameters in naturally occurring cases of canine babesiosis.

Changes in coagulation parameters were studied in dogs naturally infected with Babesia canis canis (n = 30), and haemostasis was evaluated and compared to values obtained from healthy dogs (n = 29). To date, there have not been any studies examining the dynamics of thrombin-antithrombin complex formation in cases of canine babesiosis. Coagulation parameters evaluated before (day 0) and on days 1, 2, and 3 after treatment with imidocarb (6 mg/kg inj. s.c.) included the determination of platelet counts, the formation of thrombin-antithrombin complexes (TAT), prothrombin time (PT), activated partial thromboplastin time (APTT) and antithrombin III (AT III) activity. TAT complexes were significantly elevated in animals with babesiosis on days 0 and 2 (mean 49.7 and 87.7 microg/L vs. control, 7.2 microg/L). AT III activity was significantly decreased at all time-points examined. There were no differences in PT. On days 2 and 3 the APTT was significantly shortened in the infected dogs when compared to control animals (means of 21.3 and 19.2 s vs. control, 30.0 s). Our analysis demonstrated that infected dogs had significant thrombocytopenia during the course of the study (mean day 0 - 29 x 10(9) /L, day 1 - 48 x 10(9) /L, day 2 - 47 x 10(9) /L and day 3 - 87 x 10(9) /L, vs. control -259 x 10(9) /L). These data suggest that babesiosis in dogs compromise primary and secondary haemostasis and that induction of disseminated intravascular coagulation (DIC) occurs in canine babesiosis.

Septic shock in canine babesiosis.

The records of all canine patients (86) that had been diagnosed with babesiosis and that were admitted to the Clinic for Internal Diseases, Faculty of Veterinary Medicine, Zagreb from January 2007 to December 2007 were reviewed retrospectively. All dogs that had been diagnosed with canine babesiosis and that had systemic inflammatory response syndrome (SIRS) followed by multiple organ dysfunction syndrome (MODS), and refractory hypotension, were included in this study. Of 86 patients diagnosed with canine babesiosis that were admitted during the study period, 10 had evidence of septic shock and were included in this study. Seven of the 10 dogs had a level of parasitaemia above 1%, with the highest level being 20.2%, seven of the 10 dogs were anaemic and three of the 10 dogs were leucopoenic. Thrombocytopenia was present in nine dogs. Hypoglycaemia was noted in two dogs, and bilirubinaemia in nine dogs. Four patients had involvement of two organs, five had involvement of three organs, and one had involvement of four organs. The organ that was most frequently involved was the kidney (nine cases). Central nervous system dysfunction was the rarest complication noted (one case). The mortality rate in non-septic shock canine babesiosis was 2.6%. All dogs that developed septic shock died between the first and the fourth day after admission. The 100% mortality rate that is reported here reflects the fact that in cases in which progression of the inflammatory response leads to the development of septic shock, an unfavourable outcome should be expected.

Effects of sex, age, body mass, and capturing method on hematologic values of brown bears in Croatia.

Effects of various intrinsic and extrinsic factors on 17 hematologic values from 56 brown bears (Ursus arctos) sampled in Croatia from 1981 to 2005 were evaluated. Differences between female and male bears were detected for number of erythrocytes, sedimentation rate after 30 min, and number of leukocytes and segmented neutrophils. Significant differences between free-living vs. captive and snared vs. not snared bears were detected for the same three parameters: leukocytes, segmented neutrophils, and eosinophils. It was concluded that the physical exertion of bears snared by leg, rather than their free-living status, influenced differences of results among these groups. The obtained mean values are useful reference values for the species.