First genetic characterization of equine adenovirus type 1 (EAdV-1) in Turkey

Equine adenovirus type 1 (EAdV-1) is a cause of repiratory tract infection in equids. In present study for the first time in Turkey, the prevalence of EAdV-1 in nasal swab samples obtained from horses showing respiratory symptoms was investigated by polymerase chain reaction (PCR), and molecular characterization of the hexon gene detected in the Turkish (TR) strain was performed. Overall, the prevalence of EAdV-1 was found low (1.4%) as indicated by a positive PCR reaction from the nasal swab extracts tested. Phylogenetic analysis based on the partial sequences of the hexon gene of a TR-EAdV-1 strain with those of previously isolated AdVs from different mammals and an EAdV-1 M1 strain showed that the EAdV-1 strains were placed into a unique cluster. Although the TR-EAdV-1 strain was closely related to CAV-1, CAV-2 and bat adenovirus reference strains, larger-scale studies are necessary to better understand the molecular epidemiology and population structure of EAdV-1 in Turkey.     

The influence of sex on biochemical markers of bone turnover in dogs

Biochemical markers of bone turnover have been shown to be useful as inexpensive and noninvasive tools for monitoring skeletal health. The reference range for bone markers in dogs has been set by different age groups. However, other sources of biological variations were not fully investigated in dogs. To explore whether sex influences the interpretation of bone marker data we examined serum bone markers in 33 male and 25 female dogs. The bone markers selected for this study were: bone alkaline phosphatase (BALP) and osteocalcin (OC) as indicators of bone formation, and C-terminal telopeptide (CTx) of type I collagen as marker of bone resorption. All concentrations of bone markers were lower, but still within the reference range reported for dogs. We found statistically significant differences of the median OC and CTx serum concentrations between males and females. The results of this study suggest that there are sex differences in biochemical markers of bone turnover in dogs which should be considered in interpretation of bone marker data.     

Resting and dobutamine stress test induced serum concentrations of brain natriuretic peptide in German Shepherd dogs

Studies of clinical uses of brain natriuretic peptide (BNP) represent one of the most important advances in cardiology since the introduction of echocardiography as a clinical diagnostic procedure. Defining the clinical potential of BNP in canine cardiology has not been completed yet. The aim of this study is to measure BNP concentrations in healthy German Shepherd dogs of different ages as a baseline in resting and when conventional protocol of the dobutamine stress test (DST) is applied to dogs. Concentrations of BNP were measured in blood serum by the radioimmunoassay method. The values of BNP concentrations were compared to cardiac parameters obtained by standard cardiac diagnostic procedures (radiology, electrocardiography and echocardiography). No significant differences in serum BNP concentrations existed in dogs of different ages. A statistically significant increase in BNP concentrations was registered after DST. These changes in BNP concentrations were related to ST/T electrocardiographic changes, and correlated to changes in the left ventricular internal diameter in systole (LVESD). These data suggest that BNP is not increased in aged dogs with normal cardiac systolic function and renal function, and that myocardial ischemia leads to a significant increase in BNP concentrations even in dogs with normal left ventricular function.     

Assessment of the effects of adjunctive gabapentin on postoperative pain after intervertebral disc surgery in dogs

ObjectiveTo assess the effect of adjunctive gabapentin (GBP) on pain after thoracolumbar intervertebral disc surgery in dogs.Study designProspective, randomized, controlled, clinical, ‘blinded’ trial.AnimalsSixty-three client owned dogs undergoing hemilaminectomyMethodsDogs were assigned to two treatment groups. The GBP group received gabapentin 10 mg kg^?1 orally every 12 hours starting before anaesthesia; the placebo (P) group received empty gelatin capsules. Background analgesia was initiated with intravenous levomethadone 0.6 mg kg^?1 (as the combination ‘L-Polamivet) at anaesthesia induction, followed by a fentanyl patch and levomethadone 0.2 mg kg^?1 subcutaneously every 8 hours for 24 hours. Pain was assessed by the short form of the Glasgow Composite Measure Pain Score (CMPS-SF) without the gait category, and by a Visual Analogue Scale (VAS). Serum GBP concentrations and cortisol concentrations were measured. Statistical analyses utilized chi square test, Kolmogorov-Smirnov test, two-way analysis of variances for repeated measurements, Wilcoxon test and Friedmann test as relevant. Correlations were tested by Spearman's and Pearson's correlation coefficient. p< 0.05 was considered significant.ResultsMedian CMPS-SF was lower in group GBP than in group P on days 0.5, 1, 4 and 5. However, CMPS-SF and VAS were not significantly different between groups. Both pain scores decreased significantly over time. Cortisol concentrations were not significantly different between groups. Minimum serum concentrations of GBP fell below the detection limit of 1 μg mL^?1 in 6 of 29 and 7 of 28 dogs at 24 and 72 hours, respectively.Conclusions and clinical relevance10 mg kg^?1 GBP orally twice a day did not result in a detectable reduction in pain behaviour compared to background opioid analgesia alone, although a trend to lower pain levels (p < 0.1) was present. Further studies are needed to determine if this is related to effective background analgesia or an ineffective dose of GBP.     

Chloroplast DNA polymorphism of an exotic P. mugo Turra population introduced to seaside spit of Kursiu Nerija in Lithuania

The aim of this study was to elucidate the introduction history of P. mugo in the unique landscape of the Lithuanian seaside spit of Kursiu Nerija by assessing its genetic structure and the genetic diversity. The individuals were sampled in 12 populations within an area of 3 km × 50 km along the Lithuanian part of Kursiu Nerija. P. mugo was introduced over 200 years ago to prevent sand erosion by establishing a forest cover. Chloroplast DNA polymorphism of 220 individuals of P. mugo together with 18 P. sylvestris and 11 putative P. sylvestris × P. mugo hybrids was assessed by the aid of five microsatellite markers. The standard intra-population diversity indexes were calculated. The intra-specific variation between distinct morphotypes as well as the population differentiation within the most spread P. mugo ssp. rotundata morphotype was assessed based on the haplotype frequencies by hierarchical AMOVA, G_ST/R_ST test, UPGMA clustering and PCA methods. The genetic diversity of P. mugo in Kursiu Nerija was high (H_e = 0.95; 83 different haplotypes). All except one of the P. mugo populations sampled contained a notable share of private haplotypes. AMOVA revealed high intra-specific diversity but low differentiation between the P. mugo populations. Most of the haplotypic variance was within populations. The UPGMA clustering produced groups more corresponding to the sub-species morphotypes than the geography of the populations. There was no geographical pattern of reduction in genetic diversity towards the younger plantations. A strong candidate for a species-specific DNA marker was found. After several events of introduction, the genetic diversity of P. mugo in Kursiu Nerija is very high and is structured based on the sub-species morphotypes rather than geography. The high frequency of shared and notable frequency of private haplotypes in most of the populations indicate that the major part of the P. mugo material originates from a number of geographically and genetically related sources, which more likely are introductions from abroad that the local collections. The high frequency of private haplotypes in the northernmost populations leaves a possibility for minor introductions from other genetically distinct sources. The absence of private haplotypes in one of the sampled populations indicates the use of local seed collections. The large number of shared haplotypes provides a strong evidence for a geneflow among the P. mugo taxa.     

Initial and residual efficacy of insecticides on different surfaces against rice weevil Sitophilus oryzae (L.)

The aim of the study was to investigate how various types of storage facilities with, e.g., concrete, metal, and plywood surfaces interfere with the activity of different insecticide formulations used for rice weevil Sitophilus oryzae (L.) control: malathion (EC), pirimiphos-methyl (EC), and lambda-cyhalothrin (CS and WP). Initial and residual efficacy were determined in the laboratory. Knockdown data for the initial effects were processed by probit analysis and presented as knockdown time (KDT) parameters with kdt-p lines. Delayed effects were shown as knockdown efficacy (%) determined after 24 h of weevils’ contact with 7-, 14-, 30-, 60-, 90-, 120-, 150-, and 180-day-old deposits on each surface. Malathion (EC) and pirimiphos-methyl (EC) showed the highest initial knockdown efficacy on metal, while it was 3.6 (3.4)- and 4.4 (3.3)-fold lower on concrete and plywood, respectively. Lambda-cyhalothrin (CS and WP) showed the highest initial efficacy on concrete, and slightly lower (1.3 and 2.4) fold on metal and plywood, respectively. Both formulations of lambda-cyhalothrin and malathion on metal, as well as pirimiphos-methyl on plywood were 100 % efficient against S. oryzae 180 days after the treatment. Delayed efficacy of both formulations of lambda-cyhalothrin decreased on plywood after 120 days, and after 180 days the efficacy was 55 %. All insecticides, except lambda-cyhalothrin (CS), expressed low knockdown efficacy on concrete, while the deposit of lambda-cyhalothrin (CS) on concrete was 100 % efficient during 90 days, and after 120, 150, and 180 days the efficacy was 83, 65, and 17 %, respectively.     

Deletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccine

Brucella melitensis Rev 1 is the best vaccine available for the prophylaxis of small ruminant brucellosis and, indirectly, for reducing human brucellosis. However, Rev 1 shows anomalously high rates of spontaneous dissociation from smooth (S) to rough (R) bacteria, the latter being inefficacious as vaccines. This S-R instability results from the loss of the O-polysaccharide. To overcome this problem, we investigated whether some recently described mechanisms promoting mutations in O-polysaccharide genes were involved in Rev 1 S-R dissociation. We found that a proportion of Rev 1 R mutants result from genome rearrangements affecting the wbo O-polysaccharide loci of genomic island GI-2 and the wbkA O-polysaccharide glycosyltransferase gene of the wbk region. Accordingly, we mutated the GI-2 int gene and the wbk IS transposase involved in those arrangements, and found that these Rev 1 mutants maintained the S phenotype and showed lower dissociation levels. Combining these two mutations resulted in a strain (Rev 2) displaying a 95% decrease in dissociation with respect to parental Rev 1 under conditions promoting dissociation. Rev 2 did not differ from Rev 1 in the characteristics used in Rev 1 typing (growth rate, colonial size, reactivity with O-polysaccharide antibodies, phage, dye and antibiotic susceptibility). Moreover, Rev 2 and Rev 1 showed similar attenuation and afforded similar protection in the mouse model of brucellosis vaccines. We conclude that mutations targeting genes and DNA sequences involved in spontaneous O-polysaccharide loss enhance the stability of a critical vaccine phenotype and complement the empirical stabilization precautions taken during S Brucella vaccine production.