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Amela Livnjak

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N. Hadžimusić, A. Livnjak, L. Velić, B. Čengić, D. Hadžijunuzović-Alagić

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.

N. Hadžimusiæ, V. Škapur, L. Pašiæ, A. Livnjak

Background: Llamas are becoming increasingly popular as livestock and as companion animals and their blood samples may require transportation over long distances and storage for extended periods prior to hematological analyses. Hematological parameters are influenced by a range of factors, including age, sex, diet and environment. Examining how these parameters change over time in stored blood samples can provide insights into the natural variation of these parameters in llamas, which can inform future research on the health and biology of these animals. This study sought to evaluate the stability of hematological markers at 0, 6, 12, 18, 24, 30, 36 and 42 hours of storage in blood samples of llamas (Lama glama) stored at 4°C. Methods: The study was conducted on blood from eight clinically healthy llamas captivated at Sarajevo Zoo-Pionirska dolina, Sarajevo, Bosnia and Herzegovina. The effect of storage was observed on the following hematological parameters: Red Blood Cell count (RBC), hematocrit (HCT), hemoglobin (HGB), Mean Cell Volume (MCV), Mean Cell Hemoglobin (MCH), Mean Cell Hemoglobin Concentration (MCHC), Reticulocyte count (RETIC), White Blood Cell (WBC). White blood cell differentiation, absolute white blood cell counts, platelet count (PLT) and mean platelet volume (MPV) are also included. Result: During 42 hours of storage at 4°C, it was discovered that red blood cells, hemoglobin, white blood cells, mean cell hemoglobin and reticulocyte, as well as the differentiation of white blood cells and measurement of their absolute values, remained stable. However, hematocrit increased at 12 h, while mean cell hemoglobin concentration decreased at 12 h when stored at 4°C.

Amila Šunje-Rizvan, Amina Rizvanović, Ismar Lutvikadić, A. Livnjak, Nermina Spahija, Alan Maksimović

This case report describes the remission of the fibrovascular corneal lesions in both eyes after bilateral superficial keratectomy combined with medical treatment in a German Shepherd dog. A completely blind German Shepherd dog diagnosed with chronic superficial keratitis (CSK, pannus) underwent bilateral superficial keratectomy along with subconjunctival administration of 0.1 mL of bevacizumab and mitomycin C, which where topically applied over the degenerated area of the cornea. Following the surgical procedure, cyclosporine topical ointment was given daily as life-long therapy. Notable improvements in behavior and physical activity were evident in the patient three to four weeks after the treatment. During a follow-up period over 24 months, complete remission of the fibrovascular corneal lesions was observed in both eyes, with minor seasonal conjunctival hyperemia successfully managed by administering dexamethasone and mitomycin C eye drops. This is the first report of using this specific combination of drugs during and after keratectomy in the dog, resulting in remission of CSK with no reported side effects. Superficial keratectomy, along with the administration of bevacizumab via subconjunctival injection, may present a novel approach for addressing superficial corneal neovascularization in dogs.

A. Livnjak, N. Hadžimusić

The aim of our study was to assess the values of hematological profile of Alpine goats in Bosnia and Herzegovina in order to monitor health status of goats; moreover, to determine the effect of age on hematological parameters of goats. A total of 40 apparently healthy Alpine goats included in this study were divided into four groups according to age: <1 year old (Group 1), 1 to 3 years old (Group 2), 3 to 5 years old (Group 3) and >5 years old (Group 4). The following hematological parameters were measured: Red Blood Cell count (RBC), hematocrit (HCT), hemoglobin (HGB), Mean Cell Volume (MCV), Mean Cell Hemoglobin (MCH), Mean Cell Hemoglobin Concentration (MCHC), Reticulocyte count (RETIC), White Blood Cell (WBC), as well as white blood cells differentiation and determination of absolute values white blood cells; platelet count (PLT), and Mean Platelet Volume (MPV). All values investigated in our study were within reference range, except MCHC, percentage of neutrophils and percentage of monocytes. For investigated hematologic parameters, significant (P < 0.05) age-related differences were found for HCT, MCV, MCHC, WBC, lymphocytes and platelet count. Recorded differences might be influenced by undetected infections, genetics, nutrition, fluctuations in the climate or management.

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