Dragica Stojanovic

Molecular typing and antimicrobial resistance of Salmonella Enteritidis isolated from poultry, food, and humans in Serbia.

Molecular typing and resistotyping coupled with gyrA single nucleotide polymorphism (SNP) of 60 Salmonella Enteritidis (SE) isolates originated from poultry, food, and humans in Serbia is described. Molecular fingerprinting was performed by randomly amplified polymorphic DNA (RAPD) using four primers, and the diversity index (D) was 0.688. In combination with resistotyping and gyrA SNP, D increased to 0.828. A total of 23 genetic groups were obtained. When four RAPD primers were combined, epidemic isolates from a fast-food restaurant outbreak were clustered in a distinctive genetic group. Among 60 SE strains, three had multiple resistances to three or more antibiotics. Nine strains were resistant to nalidixic acid (NAL; a non-fluorinated quinolone). The mutations in quinolone resistance-determining region (QRDR) found in NAL-resistant strains were attributed to Asp87 → Asn in six strains, Asp87 → Gly in one strain, and Ser83 → Phe in one strain. One NAL-resistant strain had no mutations in QRDR, suggesting another mechanism of resistance.

Mutational Polymorphism in the Bacterial Topoisomerase Genes Driven by Treatment with Quinolones

Overuse of antimicrobial agents in human and veterinary medicine is a serious problem worldwide. Bacteria have developed resistance mechanisms decades ago, even before wide application of antibiotics had started. Naturally they had learned to cope with an unfavourable environment well before man changed it. Bacteria can develop resistance utilizing different mechanisms. Through mobile genetic elements bacteria transfer resistance to progeny, raising concerns about not being able to control the infection (Velhner et al., 2011). An understanding of resistance mechanisms is essential for developing new therapeutics and overcoming current problems in therapy.

Biochemical parameters in rats with an applied model of sepsis (cecal ligation and puncture) with pure and mixed bacterial culture.

The goal of this work was to induce clinical sepsis in rats in order to measure the following biochemical parameters (glucoses, triglycerides, cholesterol, total proteins, albumins and creatinine). The experiments were done on 104 male Wistar rats, body weight 190 to 240 g. The rats were divided in four groups. Treated groups consisted of 28 animals while in the control groups there were 20 animals. Animals were observed and sacrificed at 12, 24, 72 and 120 hours after surgery. In this model of sepsis (cecal ligation and puncture - CLP) with pure and mixed bacterial cultures, significant changes were noticed in all biochemical parameters. Significant hypoglycemia, hypercholesterolemia, decreased concentration of urea and increased concentration of creatinine were found in the first half of the experiment in all groups of rats with sepsis (at 12 and 24 hours). Hyperproteinemia and hypotriglyceridemia reached statistically significant values at 24 h in the groups of rats in which sepsis was induced with pure bacteria culture. In the other half of the experiment significant hypoalbuminemia was found in all rats with sepsis (at 71 and 120 h). [Projekat Ministarstva nauke Republike Srbije, br. TR 31071]

UTICAJ PRODUŽENE PERORALNE PRIMENE EUGENOLA NA HEMATOLOŠKE I NEKE BIOHEMIJSKE PARAMETRE KRVI KOD PACOVA

The possible haematotoxic, hepatotoxic and nephrotoxic effects of eugenol in rats after two-week and the four-week continuous use were tested. The experiment was conducted on 72 male Wistar rats divided into six equal groups. Four were treated with different doses of eugenol (10 mg/kg bw/day, 50 mg/kg/d, 200 mg/kg bw/d and 400 mg/kg bw/d), the control group received the vehikulum (0.5% methyl cellulose, 20% propylene glycol and water), and the sixth group was the absolute, untreated control. Eugenol and the vehikulum were administered with a gastric probe, on a daily basis, for four weeks at a rate of 1 ml/100 g body weight of rats. Blood was collected by cardiac puncture of the anesthetised rats on days 14 and 28 in order to assess the haematological and biochemical blood parameters: haematocrit, red blood cell count, white blood cell and platelet counts, leukocyte formula, hemoglobin, MCV, MCH, total protein concentration, albumin, urea, creatinine, and the activity of the alanine aminotransferase (ALT), alkaline phosphatase (ALP) and creatine kinase (CK). The results showed that, having been applied for two and four weeks at doses of 10, 50, 200 and 400 mg/kg bw/day, eugenol did not show any haematotoxic, hepatotoxic or nephrotoxic effects. Eugenol administered for four weeks does not significantly affect the number of erythrocytes, haemoglobin concentration, haematocrit, red blood Ветеринарски журнал Републике Српске Veterinary Journal of Republic of Srpska (Бања Лука-Banja Luka), Вол/Vol.XIII, бр/No.2, 131–142, 2013 Milanka Jezdimirović и сар.: Uticaj produžene peroralne primene eugenola na hematološke i neke biohemijske parametre krvi kod pacova 133 cell volume, leukocyte count and leukocyte formula. Applied for two weeks it caused a significant increase in the mass of haemoglobin in erythrocytes. This effect is not specific and does not depend on the dose or the treatment duration. Eugenol in doses of 10 and 200 mg/kg/day applied for four weeks caused a significant reduction in the number of platelets, whilst the highest dose tested (400 mg/kg/day) caused a significant increase in comparison to the number of platelets in rats treated with 10 and 200 mg/kg/day. Changes in the number of platelets induced by eugenol qualitatively different and are not related to dose or length of treatment. Eugenol applied for two and four weeks does not significantly affect proteinaemia, albuminaemia, serum urea and creatinine concentrations, and serum ALP activity in rats. Three assessed doses of eugenol (10, 50 and 200 mg/kg/d) significantly decreased, whereas the highest dose increased the ALT activity. However, eugenol applied for four weeks at a dose of 200 and 400 mg/kg/d caused a statistically significant decrease in CK activity.

Hematological parameters in cows in early lactation treated with ketoprofen and their relationship with lipid mobilization and ketogenesis

Background: Dairy cows are exposed to numerous hematological and biochemical changes, what is bringing cows into the state of increased metabolic activity and physiological adaptations. These adaptive processes have resulted in increased lipid mobilization and ketogenesis with increased concentration of non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB). As a non steroidal anti-inflammatory drug, ketoprofen produces anti-inflammatory effects. The main objective of the present study was to found relationship between hematological parameters and ketoprofen administration and high lipid mobilization and ketogenesis in cows treated with ketoprofen immediately after calving. Materials, Methods & Results: Ketoprofen was used (3 mg/kg body weight) intramuscularly for three consecutive days post-partum on 15 cows of Holstein-Friesian breed. Cows of the control group (n = 15) were not treated with ketoprofen. Blood samples were collected at the day of calving, in the first and in the second week after parturition from the coccygeal vein of the both groups. Hematological parameters (erythrocytes, hemoglobin, neutrophils, lymphocytes and mean platelet volume) were determined on the automatic hematological counter. Metabolic parameters (NEFA, BHB) were determined by standard colorimetric kits using a semi-automatic biochemistry analyzer. Student’s t-test was used to determine the influence of the application of ketoprofen to the hematological parameters. The data analysis was performed using SPSS, version 19.0, software package for Microsoft Windows (IBM, Armonk, NY, USA). The results of comparison with (P < 0.05) were considered to represent statistically significant differences. Difference in correlation test between NEFA and BHB and other blood parameters in experimental and control group of cows were determined by the Fischer r-to-z transformation. The research results show that there was a increased red blood cell count and a higher concentration of hemoglobin (P < 0.05), fewer neutrophils (P < 0.01), lower number of lymphocytes (P < 0.01) and fewer mean platelet volume in cows that received ketoprofen after calving than those of the control. Determined hematological parameters and values of NEFA and BHB are in correlation. The correlation coefficients were significantly lower in the experimental group of cows. The intensity of the connection between hematological parameters and value of NEFA and BHB decreases. Discussion: It is well known that dairy cows with excessive adiposity manifest a low-grade inflammation and that elevated NEFA concentrations present positive risk factors for many proinflammatory diseases. Analyzing the results in this examination, it has been noticed that the increase of NEFA and BHB after calving were significantly lower in group of cows treated with ketoprofen. Also, determined hematological parameters and values of NEFA and BHB are in correlation. The intensity of these connection decreases in ketoprofen treated cows. In current study, in cows treated with ketoprofen there is decrease in concentrations of inflammatory mediators such as neutrophils and lymphocytes. Red blood cell and hemoglobin concentration decreased in the peripartal period as a result of inflammation. In cows treated with ketoprofen these concentrations increased. In many inflammatory conditions mean platelets volume is increased while there is a decrease in ketoprofen treated cows. Presented results indicate strong relationship between the concentration of NEFA, as well as BHB and hematological parameters indicating that the dependence of hematological parameters of intensity of lipid mobilization and ketogenesis was significantly lower in cows treated with ketoprofen immediately after calving.

Occurrence of infectious laringotracheitis on farms in Vojvodina

For over 10 years infectious laringotracheitis has occurred rarely in Vojvodina. In such situations morbidity was 40% and mortality was 16%. Drop in egg production was over 30% and after 4 weeks egg production returned to normal. After the disappearance of clinical symptoms and after 30 days of quarantine and disinfection of eggs with formaldehyde and 5% hydrogen peroxide there was no appearance of the disease in hatched chickens. Clinical observation and pathohistology results are valuable proof for diagnosis of ILT. Diagnosis of ILT should be confirmed by virus isolation or applying serological tests such as ELISA on sera from diseased chickens.