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L. Velić

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Canine pyometra is a common reproductive disorder of intact diestrual bitches. The disease is associated with inflammation and infection of the uterus leading to generalized illness. The varied reactions of the host to infection, inflammation, or trauma are collectively known as the acute-phase response (APR) and encompass a wide range of pathophysiological responses. One of these systemic responses to disease is an increase in the production by the liver of a number of serum proteins, which are known collectively as the acute-phase proteins (APP). The most important APPs in dogs are C-reactive protein (CRP) and serum amyloid A (SAA). When diagnosed with pyometra, the preferred and most effective treatment is ovariohysterectomy, a procedure that in itself causes systemic inflammation due to the tissue trauma caused by surgery. The purpose of this study was to identify the effect pyometra and ovariohysterectomy have on acute phase proteins and routinely measured biochemistry variables in serum of affected bitches. We achieved this by evaluating concentrations of CRP and SAA, as well as of some biochemistry parameters in serum before and after ovariohysterectomy in bitches with pyometra. Blood samples were collected from the cephalic vein immediately prior to and 24h, 72h and 168 hours after initiation of surgery. The levels of CRP, SAA and some biochemistry parameters - total protein (TP), albumin (ALB), globulin (GLOB), as well as activity of some enzymes: alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKLP) and lactate dehydrogenase (LDH) were determined Aims: The aim of this study was to investigate the impact of pyometra and its surgical treatment, ovariohysterectomy, on acute phase proteins, specifically C-reactive protein (CRP) and serum amyloid A (SAA), as well as on routine serum biochemistry variables in affected bitches. By evaluating the concentrations of CRP and SAA, alongside key biochemistry parameters before and after surgery, this research seeks to elucidate the physiological and biochemical responses to both the disease and its treatment, providing insight into the systemic effects of pyometra and the surgical stress response in dogs. Study Design:  The study design involved a cohort of twenty-seven female dogs, divided into two groups: seventeen clinically healthy intact female dogs serving as the control group, which were examined for routine health care or vaccinations, and ten female dogs diagnosed with pyometra at the Faculty of Veterinary Medicine, University of Sarajevo. The diagnosis of pyometra was made based on case history, clinical signs, ultrasonography, and confirmed by gross examination of the pus-filled uterus during and after ovariohysterectomy. The surgical procedure was performed using a standard mid-line approach under general inhalation anesthesia, with no post-operative complications reported. Place and Duration of Study: Faculty of Veterinary Medicine, University of Sarajevo, between June 2012 and September 2012. Methodology: Blood samples were collected from the distal cephalic vein into serum separating tubes (Vacutainer SST; Becton Dickinson, USA) for CRP and SAA measurement and serum biochemical evaluation just before the ovariohysterectomy (base values) as well as after 24 hours (day one), 72 hours (day three) and 168 hours (day seven). Blood samples from control dogs were collected only once. Serum samples were prepared by centrifugation (1.500 × g for 10 min) and stored in plain micro tubes (Eppendorf, Germany) at −20°C prior to analysis. Serum CRP concentrations in the dogs were measured with ELISA microplate reader (Anthos 2001 ELISA-reader, Anthos Mikrosysteme, Krefeld, Germany) using a commercial canine CRP enzyme-linked immunosorbent assay (ELISA) kit (Tridelta PhaseTM Range CRP Assay, Tridelta Development Limited, County Kildare, Ireland). For SAA, the analyses were performed using also a commercially available ELISA kit (Tridelta PhaseTM Range SAA Assay, Tridelta Development Limited, County Kildare, Ireland). Total protein (TP), albumin (ALB), globulin (GLOB) concentrations and alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALKP) and lactate dehydrogenase (LDH) activities were determined by using standard procedures and slides by Catalyst Dx Chemistry Analyzer, Idexx. Statistical analyses were performed using the SPSS software package (for Windows, Version 11.5, SPSS Inc, USA). All data were shown as mean ± standard deviation. The results were analyzed using the independent t-test. A P-value less than 0.05 were considered statistically significant. Results: Our results indicated significantly higher concentrations of CRP and SAA in dogs with pyometra throughout the study period compared to the control group, with CRP concentrations rising up to 15-fold and SAA up to 10-fold. Notably, these increases in CRP and SAA concentrations were mostly observed before surgery, with only limited additional increases post-surgery. This suggests that while ovariohysterectomy induces a proinflammatory response, it does not significantly escalate the levels of these proteins beyond the initial disease impact. The variability observed in CRP and SAA levels points to potential interindividual differences, impacting their diagnostic value. However, on the last postoperative day, the significantly elevated levels of both proteins in dogs with pyometra suggest a persistent, though less severe, inflammatory process, with a subsequent decline as homeostasis was gradually restored. Conclusion: The role of the acute phase response is to limit inflammation, remove the damaging factor and restore homeostasis. The systemic effects of pyometra are reflected by acute phase proteins and several biochemistry parameters.

Jelena S. Marić, D. Nedic, B. Vejnović, L. Velić, S. Obrenović

Abstract The aim of this study was to examine the seroprevalence of Leptospira spp. in dogs and red foxes in the entity of the Republic of Srpska, Bosnia and Herzegovina, after heavy rainfall and floods in 2014 and for the two years thereafter. The seroepidemiological study involved testing serum samples from dogs (n = 98) and foxes (n = 112) using MAT (microscopic agglutination test). Antibodies to at least one Leptospira spp. serovar were found in 52.04% of the tested dogs. The dog seroprevalence in 2014 (81.25%) was significantly higher than in 2015 (51.42% p <0.0001) and 2016 (22.5% p<0.05). The highest seroprevalences were for serovars Australis (76.47%), Bratislava (70.58%), Sejroe (66.67%) and Autumnalis (45.09%). Antibodies to at least one Leptospira spp. serovar were detected in 34.82% of the examined red foxes. In 2015, the fox seroprevalence was significantly higher (52.94%) than in 2016 (6.82%) (p <0.0001). The highest seroprevalences were for serovars Sejroe (64.10%), Bratislava (48.72%), Australis (43.59%) and Bataviae (25.64%). The high seroprevalence of Leptospira spp. in dogs and foxes determined during this study indicates the importance of these carnivores in maintaining leptospirosis in the study area, and the potential risk of infection for humans and other animal species that come into contact with these canids. The results obtained indicate that heavy rainfall and intense floods can result in increased Leptospira spp. infection in these canids.

Dairy cattle breeding is one of the most important branches of livestock production, which has been facing, for several decades, the chronic problem of declining reproductive performance. In 2005, the number of cattle worldwide was about 1,370,000,000, while in 2015 that number dropped below one billion, and in 2021 it shows a slight recovery as it was 1,000,970. This indicates the importance of applying different reproductive protocols in order to increase the number of cows in production. The type of bedding on which the animals stay, as well as the characteristics of the lying area itself, shows a significant impact on numerous physiological functions such as food intake, chewing, milk yield, but also levels of sex hormones. The type of bedding and lying area, which causes chronic pain and stress, leads to disorders of physiological and reproductive processes, since stress has direct negative impact on numerous cellular functions. A total of 66 dairy cows, 50 Holstein-Friesian cows kept on PD Butmir and 16 Simmental cows kept on a private mini farm, were included in the study. At PD Butmir, cows were kept in tie-stall housing system, while on a mini-farm they were kept free. Hormonal protocols of estrus and ovulation synchronization were used in April, May and June 2019. Cows were subjected to two estrus synchronization and ovulation protocols, Ovsynch and Cosynch72. At PD Butmir, 25 cows were subjected to Ovsynch and Cosynch72 protocols, respectively. At the mini-farm only Ovsynch protocol was applied. The Ovsynch protocol applied on PD Butmir had success in conception rate of 12% (n = 3), while the Cosynch72 protocol gave a score of 36% (n = 9). On the mini-farm, Ovsynch resulted in a conception of 25% (n = 4). Based on our results, the Cosynch72 protocol, compared to the Ovsynch protocol, was a better choice in the case of Holstein-Friesian cows kept in the tie-stall housing system. In the Simmental cows kept in the free stall system, the Ovsynch protocol proved to be better choice in achieving conception, compared to the Holstein-Friesian cows. Therefore, it is necessary to test several different protocols of estrus and ovulation synchronization, in order to find the most optimal one for a certain breed, type of keeping and breeding.

There is no doubt that milk and dairy products have a high nutritional value. However, their current consumption is increasingly dependent on their dietary and health properties. It is noticeable that in recent years, increasing popularity is given to those dairy products that have a positive effect on the human body. The aim of this study was to determine the fatty acid composition of sheep's milk from the area of Livno and Travnik (Vlašić), with special reference to the content of bioactive fatty acids that have a positive effect on human health. The fatty acid composition of the tested milk samples was specific due to the content of fatty acids which have been proven to have an extremely beneficial effect on human health. Concentrations of most bioactive fatty acids varied between areas, and the differences were statistically significant for arachidonic, eicosapentaenoic, docosahexaenoic, and rumenic. Milk samples from the tested areas had an almost ideal ratio of n-6/n-3 fatty acids, which makes them very favorable foods from a health point of view.

| The aim of this study was to morphologically describe the air sac system in Rosy-faced parrots (Agapornis roseicollis) and provide useful data for future veterinary care and treatment of respiratory system diseases in these pets. The research was conducted on five birds. In order to obtain the casts of the air sacs, we applied the corrosion cast technique, where we used the 26% solution of Vinylite mass. The research showed that the anatomy of the air sacs in these parrots was very similar to that in other birds, but there was some specific characteristic regarding the arrangement and connection between the air sacs. In Rosy-faced parrots we identified nine air sacs, the one unpaired, saccus clavicularis and paired saccus cervicalis, thoracicus cranialis, thoracicus caudalis and abdominalis. The casts showed that clavicular and cranial thoracic air sacs established a connection through the diverticula sternalia. These anatomical characteristics can be common to birds from Psittaciformes order and additional research need to be performed to confirm these findings.

Amina HRKOVIĆ-POROBIJA*, DVM, PhD, Assistant Professor (Corresponding author, e-mail: amina. hrkovic@vfs.unsa.ba), Aida HODŽIĆ, DVM, PhD, Full Professor, Husein OHRAN, DVM, Teaching Assistant, Nejra HADŽIMUSIĆ, DVM, PhD, Assistant Professor, Dinaida TAHIROVIĆ, DVM, MSc, Research Fellow, Almira SOFTIĆ, DVM, PhD, Associate Professor, Lejla VELIĆ, DVM, PhD, Associate Professor, Aida KUSTURA, DVM, PhD, Associate Professor, University of Sarajevo, Veterinary faculty of Sarajevo, Bosnia and Herzegovina; Vinko BATINIĆ, DVM, PhD, Teaching Assistant, University of Mostar, Faculty of Agriculture and Food technology, Mostar, Bosnia and Herzegovina Abstract The indigenous Travnik (Vlasic) cheese is produced in central Bosnia in the area of Mt. Vlasic. This cheese belongs to the group of white soft cheeses, ripened in brine under anaerobic conditions, and made from raw, thermally untreated sheep’s milk. The production technology is very simple and is adapted to mountainous conditions. Cheese is an important source of essential nutrients, in particular proteins, fat, vitamins and minerals. The aim of this study was to determine the concentration of macroelements (Ca, Na, K, Mg, P) and microelements (Zn, Fe, Cu) in 15 samples of Travnik sheep cheese using the atomic absorption spectrometry (AAS) and ultraviolet–visible spectrometry (UV-VIS) methods. A tendency of increasing Mg and K, Fe and Ca, Fe and Mg, and Fe and P was determined in the examined samples, but without statistical significance. The correlation coefficient values showed statistical validity at the level of high significance of differences (P<0.01).

A. Ćutuk, P. Bejdić, E. Šaljić, Renata Lindtner Knific, A. Dovč, L. Velić, B. Čengić

Leptospirosis is an acute, subacute and chronical contagious disease of animals and humans. Causative agents of this disease belong to the genus Leptospira, family Leptospiraca. As a disease of wild animals, leptospirosis is widespread through Europe. Certain wild animals (rodents, fox and wild boars) are important reservoirs and highly probable vectors for the spread of infection into domestic animals and humans. During the hunting season, hunting dogs are often in direct or indirect contact with wild animals that could be carriers of this disease, and the possibility of appearance and spreading within this cohort of dogs is very high. The main reasons for this study on the prevalence of Leptospirosis in Bosnia and Herzegovina are the regular contact between hunting dogs and wild animals (carriers), and the lack of dataaboutLeptospirosisinhuntingdogs.In total, 175 serum samples from 15 towns of Bosnia and Herzegovina were tested. Twelve serovars of L. interrogans were used in the microscopic agglutination test. Presence of antibodies of four serovars was confirmed. Prevalence of seropositive dogs was 15.4% (27/175). Most positive dogs had a reaction to the Pomona serovar 51.8% (n=14), while the prevalence of the Sejroe serovar was 33.3%, Icterohaemorrhagiae serovar 11.1% and Bratislava serovar 3.7%. The highest number of positive reactions 55.5% (n=15) was with serum dilution of 1:200. This study showed that most infections in dogs were caused by serovars that are currently not included in commercial vaccines. One of the most efficient preventive measure could be vaccination with the serovars most often found in wild animals, as they appear to be the most common source of the infection.

Reptiles, especially turtles, are becoming increasingly popular as pets. The haematological evaluation of turtles is an irreplaceable diagnostic tool in veterinary practice. However, the morphologic distinctiveness of turtle blood limits the use of electronic cell-counting devices, making time-consuming, manual counting techniques and evaluation of blood smears necessary. Many samples are dispatched to a laboratory over long distances, where a delay of 24 h or more may occur. At weekends, this interval may exceed 48 h. The objective of the present study was to determine the effect of storage duration at refrigerator temperature (4 °C) on the counts of red blood cells (RBC) and white blood cells (WBC), and on the mean corpuscular volume (MCV) and packed cell volume (PCV) in blood samples from healthy adult red-eared sliders. Blood samples were collected by venipuncture from the occipital venous sinus from six apparently healthy adult red-eared sliders, aged 2 to 4 years. Blood samples were analysed immediately after sampling to obtain the baseline value (BV) of the red blood cell count, white blood cell count and packed cell volume percentage. Blood was stored at 4 °C and the haematological analyses were performed after 24h, 48h and 72h. The results showed the same level of stability for RBC and WBC count, and MCV values during 72 hours of storage at 4 °C and for PCV during 48 hours. Handling of blood samples, and duration of storage of the blood samples can significantly influence the results/values of haematological tests. Consequently, the obtained values of the determined haematological parameters of improperly stored or handled blood samples can give a misleading interpretation of the results on the animal’s health status.

S. Špičić, Sanja Duvnjak, Zdelar-Tuk, K. Laroucau, Irena Reil, L. Velić, Toni Eterović, Ž. Pavlinec, Mario Šegota et al.

Silvio ŠPIČIĆ*, DVM, PhD, (Corresponding author, email: spicic@veinst.hr), Scientific Advisor, Sanja DUVNJAK, MSc, BSc, PhD, Research Associate, Maja ZDELAR TUK, DVM, PhD, Scientific Advisor, Željko PAVLINEC, BSC, MSc, Irena REIL, DVM, Assistant, Croatian Veterinary Institute, Zagreb, Croatia; Karine LAROUCAU, DVM, PhD, Scientific Advisor, Bacterial Zoonoses Unit, French Agency for Food, Environmental & Occupational Health Safety (ANSES), Maisons-Alfort, France; Lejla VELIĆ, DVM, PhD, Assistant Professor, Toni ETEROVIĆ, DVM, MSc, Scientific Associate, Veterinary Faculty University of Sarajevo, Sarajevo, Bosnia and Herzegovina; Mario ŠEGOTA, DVM, MSc, Ministry of Agriculture Republic of Croatia Veterinary and Public Health Directorate, Zagreb, Croatia; Boris HABRUN, DVM, PhD, Associate Professor, Croatian Veterinary Institute, Zagreb, Croatia; Željko CVETNIĆ, DVM, PhD, Academician, Veterinary Department Križevci, Croatian Veterinary Institute, Zagreb, Croatia Abstract In addition to zoonotic potential, Chlamydia (C.) abortus is a very important bacterium causing serious disease in small ruminants. The main outcome of the disease is abortion in the late stages of pregnancy and the economic impact for farms is significant. During a three-year period (2015–2017), 191 vaginal swabs, 24 placentas, 210 foetal organs and 2 milk samples from small ruminant abortion cases were tested for C. abortus by real-time PCR. Positive samples were detected on eight sheep farms and two goat farms, with 8.4% of total samples testing positive samples. These samples were characterised using the MLVA method, and a single MLVA genotype (genotype [2]) was identified from sheep and goat samples, suggesting highly conserved C. abortus strains among the national flock. This study is the first description of C. abortus as a causative agent of abortion in goats in Croatia. More detailed study is required to recognize the epidemiological relevance of the abortion chlamydiosis. An open register of farms with defined health status should be established for each farm at the national level for better disease(s) control.

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