* 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.

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.

Objectives This study aims to analyze human leukocyte antigen A (HLA-A), human leukocyte antigen B (HLA-B), human leukocyte antigen C (HLA-C), HLA-DRB1*, HLA-DRB3*, HLA-DRB4*, HLA-DRB5*, HLA-DQB1* loci expression in patients with rheumatoid arthritis (RA) in the Federation of Bosnia and Herzegovina. Patients and methods Deoxyribonucleic acid was isolated from peripheral blood of 48 RA patients (22 males, 26 females; mean age 36 years; range 2 to 63 years) and 104 healthy control individuals (52 males, 52 females; mean age 43 years; range 2 to 76 years). Deoxyribonucleic acid samples were analyzed using polymerase chain reaction-sequence-specific primers and sequence specific oligonucleotides methods. Results The most frequent allelic groups in RA patients were HLA-DRB1*01 (odds ratio=2.795; 95% confidence interval: 1.441-5.421; p=0.004) and HLA-DRB1*04 (odds ratio=2.573; 95% confidence interval: 1.214-5.453; p=0.023). Among RA patients, the most frequent genotype for the allelic group HLA-DRB1*, in the light of the common epitopes theory, was observed for DRB1*01/DRB1*13. This genotype indicates an increased incidence and relative risk (odds ratio=11.09). Conclusion The most common genotype in our RA patients was DRB1*01/DRB1*13, which showed increased frequency and a high relative risk. This genotype variant may be considered a predisposing factor for the development of RA.

Abstract Adaptation mechanisms as response to water content, oxygen level and pollutants are very important and they can be interpreted by hematological analysis. The aim of this study was the analysis of hematological and immune adaptations of common carp (Cyprinus carpio Linnaeus, 1758) to thermal stress. All specimens were divided into a control and experimental group. The control group of fish was exposed to a constant water temperature of 10°C. We induced thermal stress in experimental fish by gradually heating water to 28°C, held for 30 minutes and then comparing the obtained results with the control fish. Short-term hyperthermia lead to an increase of the number of leukocytes, especially pseudoeosinophilic granulocytes and monocytes, while the number of neutrophils and lymphocytes was reduced. The analysis of the leukocyte number and differential blood count in the control group showed high individual variation of segmented granulocytes, monocytes and pseudoeosinophilic granulocytes. Statistically significant differences (p=0.00) were found for the white blood cells, nonsegmented neutrophils and pseudoeosinophils between the control and experimental group. The experimental group of males had an increased number of white blood cells, monocytes and pseudoeosinophils, where significant differences were found for nonsegmented and total neutrophils and also for pseudoeosinophils (p=0.00), lymphocytes (p=0.01) and monocytes (p=0.03). Females had an increased total number of white blood cells, lymphocytes, monocytes and pseudoeosinophils, while significant differences (p=0.00) were obtained in the number of white blood cells, nonsegmented and total neutrophils and pseudoeosinophils between the control and experimental group. Adaptation mechanisms in carp after water temperature heating are mostly reflected in the increase of pseudoeosinophils and the decrease of neutrophils.

AIM: The research was conducted by genotyping two Human Leukocyte Antigen (HLA) gene classes. The main objective of this research was to investigate distribution and frequency of the allelic groups, genotypes and haplotypes in the gene loci of HLA class I (HLA-A*, -B*, -C*) and HLA class II (HLA-DRB1*, -DQB1*) in patients included in the program of cadaveric renal transplantation. MATERIAL AND METHODS: Our study covered 186 blood samples of patients who are registered on the list for cadaveric renal transplantation in Federation of Bosnia and Herzegovina and included 59 control, healthy unrelated individuals. For the HLA typing, we have used three different methods: micro lymphocyte cytotoxicity test (MLCT), Polymerase Chain Reaction (PCR) – Sequence Specific Primers (SSP) and PCR – Sequence-Specific Oligonucleotides (SSO) or Luminex technology. All patients and cadaveric donors were tested using the three methods because the system is polymorphic. RESULTS: Analysis of the results of genotyping HLA class I gene loci identified dominant HLA-A*02, HLA-B*35, HLA-C*07 allelic groups. Analysis of the HLA class II gene loci genotyping showed that HLA-DRB1*11 and HLA-DQB1*03 loci had the highest incidence in HLA class II. CONCLUSION: Based on our results and previous research, there were no observed differences between allelic frequencies and genotypes of healthy people and people with ESRD. Differences between allelic groups occurred, but they were not statistically significant, except HLA-C*01 (p = 0.020).