The main circulatory medium of echinoderms is the coelomic fluid. Biochemistry of coelomic fluid is very complex and slightly different from seawater. The main aim of this research was to analyse concentration of electrolytes (potassium, calcium, magnesium, chlorine and sodium), heavy metals (lead, chromium, cadmium and cobalt) and iron in coelomic fluid of sea urchin, Arbacia lixula as an indirect indicator of seawater contamination. After precise statistical evaluation it was observed that electrolyte concentrations were; Na 189.20±11.41 mmol/l, Cl 134.06±37.08 mmol/l, Mg 4.24±1.08 mmol/l, Ca 3.04±0.84 mmol/l. Biochemical content of heavy metals in coelomic fluid was; Co 1.292±0.879 ppm, Pb 0.644±0.131 ppm, Cr 0.116±0.055 ppm, Cd 0.031±0.017 ppm and iron 0.259±0.058 ppm, and it depends on the potential accumulation level in the organism. The composition of electrolyte and heavy metal content varies depending on the composition of seawater. Obtained values are within the range of values most commonly found in low polluted areas of the Adriatic sea. Similar models may be applied for monitoring of heavy metal contamination in marine environment.

Thermal changes in water cause many metabolic changes that manifest themselves in physiological fish adaptations. The analysis of hematologic and biochemical blood parameters provides important information on environmental influences on the health status of fish. The hematocrit (HCT) (l/l), hemoglobin concentration (Hb) (g/l), mean corpuscular volume (MCV) (fl), mean corpuscular hemoglobin (MCH) (pg), mean corpuscular hemoglobin concentration (MCHC) (g/l) and red blood cells (RBC) (x1012/L) were analyzed. Animals were grouped into two groups: control (n=10) and experimental (n=16). The experimental fish were exposed to 28°C for 30 min. Puncture of the heart was done and the blood without anticoagulant was analyzed. During hyperthermia, an increase in hematological parameters was observed, except for MCV values that were low. Significant differences were established only for the number of erythrocytes and the hematocrit values (p<0.05). The results showed a decrease in MCV and an increase in the value of other erythrocyte parameters. Significant changes in the number of erythrocyte and hematocrit values were found. Some hematological parameters such as erythrocyte and MCV values are significant stress indicators and can serve us as important factors for physiological adaptations of fish. The carp shows excellent ability to adjust to temperature variations that can be seen through the analysis of hematological status.

Abstract Emperipolesis is considered a physiological phenomena often present in various pathophysiological conditions, but its etiology is still unknown. In this study, we analyzed the number of megakaryocytes and the percentage of emperipoletic cells in the sternal and femoral bone marrow of Wistar rats. Five types in the thrombopoiesis lineage (megakaryoblasts, promegakaryocytes and megakaryocytes - acidophilic, basophilic and thrombocytogenic) were determined. Except for basophilic megakaryocytes, significant differences were found for number of thrombopoietic cells in the sternal and femoral bone marrow. A larger number of thrombocytogenic megakaryocytes were present in the sternal bone marrow. Emperipoletic cells were significantly present in the femoral compared to the sternal bone marrow. Emperipolesis was typical for lymphocytes and neutrophils individually, while emperipolesis with two or more cells within thrombopoietic cell was also present (1-7 %) and significant differences between the sternal and femoral bone marrow were detected. Emperipolesis was found in all analysed rats and it most commonly occured within mature megakaryocytes and rarely megakaryoblasts, while it was not recorded in the promegakaryocytes. The high incidence of megakaryocytes with emperopolesis in rats could be a consequence of “normal” cell retention in the cytoplasm of megakaryocytes while passing blood cells to circulation or related to haematopoietic response due to high incidence of inbreeding.

* Correspondence: suljevic.damir@gmail.com A great success in the evolution of animal behavior and physiology is the ability to survive seasonally unfavorable conditions, especially when it comes to variable and extremely low temperatures (Roots, 2006). Environmental variations lead animals to a state of hibernation, which causes depression of metabolic and physiological processes, as well as a slow development and reproduction so that the animal can survive (Dugbartey and Henning, 2013). Life activities of the gastropods depend on environmental factors throughout the whole year. Shells have great importance because they reduce water loss during summer days. The formation of the calcareous epiphragm, with which they bind to substrates, is an additional protection mechanism of evaporation (Elmslie, 1998). Snail hibernation includes shell closing by the secretion of the epiphragm, while energy is gained from stored food reserves during this process (Holtz and Von Brand, 1940). In experimental conditions, aestivation can last for years, which suggests that the metabolic rate depends on both exogenous and endogenous factors (Nowakowska, 2011). Respiration is reduced during hibernation and about 10% of the fluid is lost, which affects heart rate, oxygen consumption, and energy requirements (Nicolai et al., 2011). Metabolic changes during hibernation affect the value of circulating hemocytes. The phagocytic activity of hemocytes and lectins, as a defense mechanism, takes place thanks to the presence of calcium ions in the snail hemolymph. Consequently, the changes in the values of calcium ions affect the hemocyte number (Saleddin and Wilbur, 1984). Many studies confirmed the importance of hemocytes in immune responses, antibody recognition, phagocytosis, encapsulation, and cytotoxic reactions (Yoshino et al., 2001). Two types of hemocytes, granulocytes and hyalinocytes, have been identified in the hemolymph of Biomphalaria (Ratcliffe, 1985). Seasonal factors and different experimental tests affect the total number, type, and behavior of hemocytes (Oliver and Fisher, 1995). The role of hemocytes today is most widely considered as an immunological response to infections. Seta et al. (1996) reported that the phagocytic potential of hyalinocytes is questionable and that only granulocytes are crucial in the immune response. Vinaud et al. (2008), Delgado et al. (2001), and Matricon-Gondran and Letocart (1999) identified four types of hemocytes based on their size (small, medium, large, and gigantic cells) in the hemolymph of Biomphalaria glabrata. According to morphological criteria, there are two types, round cells and spread cells (Sminia, 1981), while in the study of Adamowicz and Bolaczek (2003), they refer to these cells as type I and type II. Wojtaszek et al. Abstract: The total number of hemocytes and their morphological characteristics were analyzed in Helix pomatia Linnaeus, 1758, by light microscopy during different phases of hibernation. Three types of hemocytes were identified: hyalinocytes, agranulocytes, and granulocytes. Total number of hemocytes decreased significantly during hibernation. Most intensive changes in cell size, morphology, and appearance of special structures in granulocytes happened during the hibernation period itself. Termination of hibernation coincides with very important granulocyte transformation and the appearance of specific gigantic short-lived cells. All of the changes in cell number and morphology are characteristics of different hibernation phases and have a major role in adaptive hypothermia. Granulocytes are hemocytes of great importance in immune response while specific functions of hyalinocytes and agranulocytes remain largely unexplored.

Total number and types of hemocytes were analysed in hemolymph of Roman snail (Helix pomatia L.). Significant differences in the number and types of hemocytes during the season at different locations were identified. The average number of hemocytes ranged from 154 to 288 per 10 μL of hemolymph. The highest number of hemocytes was recorded in snails at higher altitudes. Significant intergroup differences between all sites were identified. The most numerous hemocytes were granulocytes, followed by agranulocytes, while hyalinocytes were the least common in the post-hibernation period. Post hoc analysis showed certain intragroup differences for most locations except for comparison between wet habitat and habitats with anthropogenic influence, and between wet habitats and urban place with wet vegetation. A large number of transformed hemocytes in the encapsulated phase were identified in snail hemolymph infected with flat worms. Our results represent the first record of data for the number of hemocytes in molluscs and the first data for hemocytes of Helix pomatia. Further investigations and development of a uniform classification of hemocytes and their role in the immune response would be necessary in the future.

In this study we analyzed the effects of NaOCl solution on the blood and biochemical parameters of the rainbow trout Oncorhynchus mykiss (Walbaum, 1792). The treatment lasted for three days with prior dechlorination of the water. Significant differences in the concentration of K+, Ca2+, Cl-, proteins, glucose were detected and the largest variations were found for creatinine concentration (P<0.05). Significant differences were found Hct, Hb concentration, RBC, WBC, MCH and MCHC values, percentage ratio of lymphocytes, segmented granulocytes and the total number of neutrophil granulocytes (P<0.05). Small doses of NaOCl alter the acid-base balance, suggesting a very low-level adaption in the rainbow trout.

BACKGROUND: AFP serum levels are considered as diagnostic and specific for hepatocellular carcinoma (HCC) in patients with liver cirrhosis (LC). AIM: This study aimed to examine the diagnostic value of AFP in the distinguishing of patients with HCC from patients with LC, and to analyse the potential correlation between AFP levels and liver disease stages. MATERIAL AND METHODS: Fifty patients with LC and fifty patients with HCC were included in this study. The majority of the patients were males, while the HBV aetiology was dominant. RESULTS: Significant differences between LC and HCC patients were detected for AST, ALT, GGT, bilirubin, AFP and AP. Patients with HCC had higher AFP values compared to LC. There was no significant correlation between the size of the tumour lesion and serum AFP levels. A positive correlation between AFP concentration and GGT activity was determined, as was the negative correlation between AFP and age of the subjects. The AFP value of 23.34 ng/m showed high sensitivity (84%) and specificity (82%). CONCLUSION: The size of the surface below the ROC curve (AUC) was 0.877 (0.80-0.95), which makes AFP a good biomarker and this diagnostic test is sufficient to separate patients with HCC and LC.

Trivalent chromium is an essential trace element for humans and animals and it is the final product of hexavalent chromium reduction in the cell. It is an integral part of the glucose-tolerance factor, which increases the systemic impact of insulin. The aim of this study was to analyse the effect of trivalent and hexavalent chromium on selected biochemical parameters in Japanese quails. Birds were divided into three groups: group I (control group, n=12), group II (treated with trivalent chromium, n=12) and group III (treated with hexavalent chromium, n=12). During 20 days of follow-up, control and experimental groups of quails had identical diet; chromium was added daily into the food (5 mg/kg) and water (5 mg/L) of the experimental groups. Glucose concentrations were the highest in Group III, while lowest glucose concentrations were obtained in Group I. There were higher differencies regarding blood biochemical parameters between control group and quails treated with Cr(VI). Our results showed that hexavalent chromium had negative effects on biochemical parameters in quails, while treatment with Cr(III) showed minor fluctuations in concentrations or activities of blood biochemical parameters.

Several haematological parameters in Herzegovinian donkeys based on gender and age were analysed in this study. The study included 30 clinically healthy donkeys (12 males and 18 females), between one and twenty years of age. Donkeys were grouped in three age classes: young (1-2 years, n=10), adult (3-6 years, n=12) and old donkeys (7-20 years, n=8). A total of seven haematological parameters were analysed. In both genders and in all age groups, the mean values of MCHC were slightly above the recommended reference intervals for donkeys in general. The mean values of HCT in females and in adult donkeys were lower than the reference intervals. The PLT mean values in female, and in young and old Herzegovinian donkeys were lower than the reference intervals. Gender significantly affected HCT and Hb values (p=0.049; p=0.046, respectively), which were significantly higher in males. Age significantly affected WBC and LYM count, which were lowest in young donkeys.

Abstract Urinary tract infections can be uncomfortable, painful and even dangerous for most dog breeds. Clinical signs are often nonspecific and insufficient for diagnosis. Urinalysis in combination with biochemical parameters and urine culture is the best combination of clinical findings for diagnosis of urinary tract infections. The incidence of urinary tract infections in dogs population is growing and 27% of dogs develop an urinary tract infection through their life. Urinary infections occur more often in the elderly than in younger dogs. More than 70% of all urinary tract infections are infections with one bacterial species. Biochemical profile is important aspect for diagnosis establishment, but due to the nature of action infection by different agents may be considered as individual case. The main aim of this research was to analyse biochemical parameters of Maltese and Belgian Shepherd (Malinois) dog breed, who were affected by urinary tract infections. Urea concentration was elevated in Malinois, while urea, phosphates, albumins and alkaline phosphatase activity were elevated in Maltese dogs. Statistical analysis showed differences in concentrations of urea, creatinine, phosphates, so as alanine aminotransferase, alkaline phosphatase and amylase activity between compared breeds during acute urinary infections. Maltese dogs are less resistant to bacterias, that causes urinary tract infections, and have lower chance to maintain homeostasis of biochemical parameters in blood during urinary bacterial infections, in comparison to Maltese dogs.