Loggerhead sea turtle Caretta caretta plasma biochemistry and proteome profile modulation during recovery.

The aim of the study was to monitor and analyse injured and diseased loggerhead sea turtles (Caretta caretta) plasma proteome profiles and biochemistry parameters during their recovery period in rescue centre within different age and recovery period groups, and determine the potential biomarkers that can be used in diagnostics. The plasma biochemical parameters of total protein and glucose content, accompanied by aspartate aminotransferase (AST) and N-acetyl-cystein-activated creatinine kinase (CK-NAC) are highlighted as valuable and potential biomarkers of turtle's health status and condition. Using high throughput tandem mass tag (TMT)-based proteomic approach we identified 913 plasma proteins, 12 of which shown to be modulated in loggerheads age groups, and identified as a part of (i) platelet degranulation, (ii) neutrophil degranulation, and (iii) innate immune system pathways. The neurofascin (NFASC) is shown to be differentially abundant among all the age groups, and alpha-1-acid glycoprotein 2-like (ORM2) and alpha-1-antitrypsin-like (SERPINA1) proteins were recognized as members of all three above mentioned REACTOME pathways. Furthermore, 29 of plasma proteins were significantly differentially abundant in loggerheads age and recovery period groups. Out of 15 recognized pathways, those proteins were mostly included in three specific REACTOME pathways: (i) post-translational phosphorylation, (ii) regulation of Insulin-like Growth Factor (IGF) transport and uptake by Insulin-like Growth Factor Binding Proteins (IGFBPs), and (iii) platelet degranulation. The alpha-fetoprotein (AFP) was the only protein which showed statistically significant up-regulation patterns in all loggerhead age groups before release from the rescue centre, and the complement component 3 (C3) protein was the only protein modulated in all recovery period groups. Furthermore, C3 protein takes part in 9; and followed up with apolipoprotein A-I (APOA1) in 7; complement component 4 (C4), complement component 5 (C5) and kininogen-1 (KNG1) in 6 REACTOME pathways. Thereby, those proteins are highlighted and recommended as potential biomarkers of turtle's health status. Data are available via ProteomeXchange with identifier PXD029569. Finally, based on our results, we believe that comprehensive omics approach and routine plasma biochemical analysis, accompanied by proteins of acute phase, acid-base status and immune-response indicator analysis may significantly and reliably improve assessment of captive loggerheads rehabilitation and medication. SIGNIFICANCE: Monitoring and comparison of loggerhead sea turtles (C. caretta) blood plasma biochemistry parameters and plasma proteome profiles in relation to the age, and recovery period pointed out significantly differentially abundant proteins, along with certain biochemical parameter contents as potential biomarkers of turtle's fitness, health status and physiology.

Changes in saliva proteins in cows with mastitis: A proteomic approach.

This study aimed to evaluate the possible saliva proteome changes in cows with mastitis using a Tandem Mass Tags (TMT) proteomics approach. For this purpose, the salivary proteomes from healthy cows and cows with mastitis were analysed, and their serum proteomes were also studied for comparative purposes. A total of eight saliva and serum paired samples for each group were used for the proteomic study, and eight additional samples for each group were analysed in the analytical and overlap performance studies. In saliva samples, 2192 proteins were identified, being sixty-three differentially modulated in mastitis. In serum, 1299 proteins were identified, being twenty-nine differentially modulated in mastitis. Gamma glutamyl transferase (γGT) in saliva and serum amyloid A (SAA) were validated by commercially available automated assays. In conclusion, there are changes in protein expression and metabolic pathways in saliva and serum proteomes of cows with mastitis, showing different response patterns but complementary information.

Identification of changes in serum analytes and possible metabolic pathways associated with canine obesity-related metabolic dysfunction.

The main objective of this study was to identify analytes that could change and that could help to clarify the metabolic and physiopathological changes related to canine obesity-related metabolic dysfunction (ORMD). For this, serum from 35 overweight/obese dogs, with and without ORMD, was submitted to a comprehensive panel of biochemistry analysis, a gel-free tandem mass tag isobaric label-based proteomic analysis, and, finally, selected proteins were used as a starting point for creating a protein interaction network. Dogs with ORMD showed significantly higher serum concentrations of alanine aminotransferase (ALT), alkaline phosphatase (ALP), Ca, total proteins, albumin, total cholesterol, triglycerides, glucose, and butyrylcholinesterase (BChE) activity in comparison with dogs without ORMD. Proteomic analysis revealed that 23 proteins related to lipid metabolism, the complement factor system, cellular adhesion and functionality, inflammation, and coagulation were altered in dogs with ORMD. Finally, the obtained protein interaction network highlighted that the central term of this network was the negative regulation of the immune response. These data suggest that canine ORMD is associated with changes in analytes that reflect altered lipid metabolism, and liver and immune function impairment and suggests the potential for a prothrombotic state and lung function alterations.

Plasma markers of inflammation and hemostatic and endothelial activity in naturally overweight and obese dogs.

BACKGROUND: Obesity is one of the most prevalent health problems in the canine population. While haemostatic parameters and markers of endothelial function have been evaluated in various disease conditions in dogs, there are no studies of these markers in canine obesity. This study was designed to evaluate the effect of naturally gained weight excess and obesity on inflammatory, hemostatic and endothelial biomarkers in dogs. A total of 37 overweight and obese dogs were compared with 28 normal weight dogs. RESULTS: Overweight and obese dogs had significantly elevated concentrations of serum interleukin-6 (IL-6) and C-reactive protein (hsCRP). Number of platelets, activity of factor X and factor VII were significantly higher, while activated partial thromboplastine time (aPTT) and soluble plasminogen activator receptor (suPAR) were significantly decreased. Statistical analysis of high mobility group box - 1 protein (HMGB-1), soluble intercellular adhesive molecule -1 (sICAM-1) and plasminogen activator inhibitor type 1 (PAI-1) concentrations did not show significant differences between the total overweight and obese group and the normal weight group of dogs. CONCLUSIONS: Analytical changes in the dogs in our study reflects that weight excess in dogs can be associated with a chronic low degree of inflammation and a hypercoagulable state, where primary and secondary hemostasis are both affected. However obesity is not associated with impairment of endothelial function in dogs.

Plasma biomarkers of SIRS and MODS associated with canine babesiosis.

Canine babesiosis is a tick-borne disease caused by the haemoprotozoan parasites of the genus Babesia. Early detection of systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) is of major importance in clinical practice for providing information about severity and outcomes of the disease and therapy. Plasma samples were taken at admission from five dogs with uncomplicated babesiosis caused by B. canis canis, five dogs with babesiosis and SIRS, five dogs with babesiosis and MODS, and five healthy dogs. After two-dimensional electrophoresis and capillary reversed - phase liquid chromatography coupled online with tandem mass spectrometry, 68 differentially expressed spots with level of significance P<0.05 were detected between groups. SIRS in babesiosis was characterised by increases in paraoxonase 1 and apoA-I, whereas MODS with decrease of complement inhibitors leading to prolonged complement activation and decrease of vitamin D binding protein due to haemolysis and activation of the coagulation cascade.

Relationship between paraoxonase 1 activity and high density lipoprotein concentration during naturally occurring babesiosis in dogs.

Paraoxonase 1 (PON1) is a negative acute phase protein bound to high density lipoproteins (HDL) and during the acute phase response (APR) protects HDL from peroxidation. The aim of this study was to assess the relationship between PON1 and HDL in canine babesiosis, a disease characterized by oxidative damages and by an APR. PON1, HDL and C-reactive protein (CRP), were measured in blood collected from 15 controls and 29 dogs with babesiosis sampled at admission, and on days 1 and 7 after treatment. At admission, PON1 and HDL were significantly lower in affected dogs. HDL concentration increased at day 1 while PON1 increased and CRP decreased at day 7. This suggests that the decrease of PON1 at admission is in part due to an increased consumption, the decreased HDL may depend on lipid peroxidation and its rapid increase after treatment may depend on the antioxidant activity of PON1.

Markers of coagulation activation, endothelial stimulation, and inflammation in dogs with babesiosis.

BACKGROUND: Babesia infections in dogs can result in a wide range of clinical and laboratory presentations, including coagulopathy. Expression of intercellular adhesion molecule-1 (ICAM-1) and von Willebrand factor (vWF) in dogs with babesiosis is unknown. OBJECTIVES: Whether inflammation in babesiosis triggers activation of ICAM-1 and the coagulation system. ANIMALS: Twelve and 10 dogs with naturally occurring babesiosis before and after antiparasitic treatment, respectively, were compared with 10 healthy dogs. METHODS: In this prospective study, diagnosis was made by blood smear examination and confirmed by PCR. C-reactive protein (CRP), soluble intercellular adhesion molecule 1 (sICAM-1), and von Willebrand factor (vWF) levels were measured by a canine ELISA kit, fibrinogen (FIB) and factor VIII activity levels were measured by coagulometric methods, and blood cell counts (WBC, RBC, PLT) were determined with an automatic analyzer. RESULTS: Compared to healthy dogs, the CRP, sICAM-1, and FIB concentrations were significantly increased before therapy and remained high for 3 days after therapy in dogs with babesiosis. vWF activity was significantly decreased in dogs with babesiosis before treatment. FVIII activity did not differ between dogs with babesiosis and healthy dogs. WBC; RBC and PLT were significantly lower before treatment and normalized by 3 days after treatment. CONCLUSION AND CLINICAL IMPORTANCE: A proinflammatory condition in babesiosis appears to influence endothelial dysfunction and hemostatic activity. Although clearly beneficial for the parasite, sequestered blood cells can obstruct blood flow in small vessels, promote an inflammatory state, and could increase the severity of babesiosis.

Co-oximetry in clinically healthy dogs and effects of time of post sampling on measurements.

OBJECTIVES: Co-oximetry is a complex and valuable laboratory method that measures haemoglobin species and oxygenation status by multi-wavelength spectrophotometry. The purpose of this study was to establish reference intervals for clinically healthy dogs and to determine the effect of time of analyses and sex of animals on the accuracy of results. METHODS: Blood was collected from 27 healthy adult dogs of various breeds and sex. Co-oximetry was performed on a CCX co-oximeter that measures eight haemoglobin and oxygen transport related parameters: carboxyhaemoglobin (COHb), deoxyhaemoglobin (HHb), oxyhaemoglobin (O(2)Hb), methaemoglobin (MetHb), total haemoglobin (tHb), oxygen saturation (SO(2)%), oxygen content (O(2)Ct) and oxygen capacity (O(2)Cap). RESULTS: Results obtained after 2 and 4 hours were not significantly different from those obtained immediately after sampling. But after 48 hours, the results for total haemoglobin, oxygen saturation, oxyhaemoglobin, oxygen content and oxygen capacity were significantly lower, and carboxyhaemoglobin and deoxyhaemoglobin values were significantly higher than determination immediately after sampling. Gender had no significant impact on co-oximetry values. CLINICAL SIGNIFICANCE: Co-oximetry offers several advantages compared with other methods, including ease of use, increased accuracy and greater differentiation among haemoglobin species.

Poisoning of a dog with the explosive pentaerythrityl tetranitrate.

A three-year-old male Labrador retriever was presented at the Clinic of Internal Medicine, University of Zagreb, Croatia. The owner reported that the dog was ataxic, and this was evident by its markedly unsteady, swaying gait. The dog also had difficulty rising and fell several times while trying to stand. It had come into contact with the explosive, pentaerythrityl tetranitrate, while training to detect explosives. The following clinical symptoms were observed: bradycardia, depression, mild disorientation and a broad-based stance. The dog had conscious proprioceptive deficits in the hindlimbs, but cranial nerve function was normal except for miosis. Ion scan analysis of the dog's serum after evaporation of the current phase by mass spectroscopy revealed the presence of fragments that are characteristic of pentaerythrityl tetranitrate. The aim of the present case report was to identify pentaerythrityl tetranitrate poisoning and describe the clinical signs of pentaerythrityl tetranitrate poisoning in dogs. To the authors' knowledge, there are no published scientific articles on pentaerythrityl tetranitrate poisoning in dogs.

A seroepidemiologic survey of canine visceral leishmaniosis among apparently healthy dogs in Croatia.

Cross-sectional investigation was done on seroprevalence of Leishmania sp. infection among apparently healthy dogs in an area where canine leishmaniosis is endemic. Survey included 68 dogs living in the coastal city of Split, and 238 dogs living in 12 villages scattered in the hinterland. Each dog was clinically examined for the presence of some discrete signs compatible with leishmaniosis and by dot-ELISA modification determined the presence of anti-Leishmania antibodies. The titre 1:600 and higher was regarded as positive in the study. The seroprevalence ranged from 0 to 42.85%, depending on the location. 54.34% of the seropositive dogs had moderately enlarged lymph nodes and/or some discrete changes on the skin. In our parasitological study, Leishmania sp. was isolated from several seropositive animals that had some clinical signs and from a few which did not have any. Data analysis revealed that serological positivity to Leishmania sp. was not associated with a dog's outdoor lifestyle and utility, but was associated with the gender and age.

Effect of different daily doses of gizzerosine on the serum 1,25-dihydroxycholecalciferol concentration in laying hens.

Five groups of laying hens were treated with different gizzerosine doses (0, 2.5; 5.0; 7.5; 10.0 mg/kg/body weight of gizzerosine) daily over a 21-day period to determine the serum concentrations of 1.25-dihydroxycholecalciferol (1,25(OH)2D), total calcium, inorganic phosphorus and magnesium. Blood samples were taken on days 7, 14, and 21 of the experiment. The concentration of 1,25(OH)2D remained unchanged after day 7 in the gizzerosine-treated birds compared to the control group. After 14 days, it was significantly lower in the birds receiving. gizzerosine, compared with the control group. On day 21, 1,25(OH2)D concentrations were also significantly decreased in all 4 gizzerosine-treated groups compared with the control hens. The serum total calcium, inorganic phosphorus and total magnesium concentrations varied significantly, but irregularly, during the period of the study.

Antibodies to spotted fever group rickettsiae in dogs in Croatia.

The prevalence of IgG antibodies to spotted fever group rickettsiae was studied by the indirect immunofluorescent antibody assay in a sample of 194 canine sera from different parts of Croatia. Dogs (n = 141) from urban and suburban-semirural environs from the middle part of the eastern coast of Adriatic sea, and dogs (n = 53) from Zagreb, the capital of Croatia located in the northern continental part of Croatia, were tested. The total of 88 (62.4%) dogs from southern coastal Croatia had antibodies to Rickettsia conorii, with a significantly higher prevalence (69.9%) in suburban-semirural areas. The prevalence rate of antibodies to Rickettsia conorii in canine sera from urban environment in the north of Croatia was 20.7%. The results show that the members of spotted fever group rickettsiae are present in both coastal and continental Croatia.

Effects of histamine on gizzard erosions and on the activity of selected enzymes in chickens.

Dose- (5, 10 and 15 mg/kg body mass) and time-dependent (2, 6, 12 and 24 h after treatment) effects of histamine on gizzard lesions and serum AST, ALT and CK activities of chickens are reported. Morphometric results and histopathological examination revealed that the most effective histamine dose which induced severe gizzard lesions was 10 mg/kg b.m., especially 2, 6 and 12 h after administration. No difference from the control values was found after 24 h. That dose also induced an elevation of serum enzyme (AST, ALT, CK) activities, which was most expressed for the activity of ALT.