Identification of a DNA nonhomologous end-joining complex in bacteria

In eukaryotic cells, double-strand breaks (DSBs) in DNA are generally repaired by the pathway of homologous recombination or by DNA nonhomologous end joining (NHEJ). Both pathways have been highly conserved throughout eukaryotic evolution, but no equivalent NHEJ system has been identified in prokaryotes. The NHEJ pathway requires a DNA end-binding component called Ku. We have identified bacterial Ku homologs and show that these proteins retain the biochemical characteristics of the eukaryotic Ku heterodimer. Furthermore, we show that bacterial Ku specifically recruits DNA ligase to DNA ends and stimulates DNA ligation. Loss of these proteins leads to hypersensitivity to ionizing radiation in Bacillus subtilis. These data provide evidence that many bacteria possess a DNA DSB repair apparatus that shares many features with the NHEJ system of eukarya and suggest that this DNA repair pathway arose before the prokaryotic and eukaryotic lineages diverged.     

Adaptive selection of foraging and nesting habitat by black kites (Milvus migrans) and its implications for conservation: a multi-scale approach

Black kites (Milvus migrans) are vulnerable and in decline within Europe. Here, we investigate selection of foraging and breeding habitat in a high-priority population in the Italian pre-Alps. Compared to a random distribution, kites foraged preferentially near water, over extensively managed grassland and within 1 km of nest-sites. Urban areas were positively selected near lakes but otherwise avoided. Foraging performance was higher over water than over terrestrial habitats. Kites nested on cliffs and trees and preferentially near water, far from paths and villages and in rugged and steep micro-sites. Tree-nests were located in the most mature tree in the stand. Productivity was positively related to the availability of water bodies. Therefore, food availability and human disturbance/persecution limited foraging and breeding performance. Guidelines to maintain or enhance current population levels include: (1) setting up reserves covering 10% of the areas within 1 km of large lakes; (2) converting current derelict coppice-woodland to high forest; and (3) enhancing subsidies for extensively managed grassland. Our results highlight the importance of cross-scale models integrating selection of foraging and breeding habitat and reinforce the importance of the spatial configuration of key resources for more realistic conservation management in mosaic landscapes.     

Left basilar systolic murmur in retired racing greyhounds

Nineteen of 28 (67%) Greyhounds enrolled in the Blood Donor Program at The Veterinary Teaching Hospital, The Ohio State University (Columbus, OH), had a left basilar systolic murmur. Ten Greyhounds with murmurs and 9 without murmurs were evaluated to gain knowledge about the pathogenesis of this murmur. Echocardiograms were performed without sedation by means of a GE Vivid 7 Echocardiographic System with a continuous ECG; systolic arterial blood pressure (SABP) was measured with an Ultrasonic Doppler Flow detector model 811-B. The mean peak aortic velocity in the Greyhounds with murmurs (2.15 m/s; range, 1.8-2.2 m/s) was significantly higher than in the Greyhounds without murmurs (1.89 m/s; range, 1.6-2.0 m/s) (P < .001); there were no significant differences between groups for aortic valve or annulus diameter, fractional shortening, pulmonic velocity, SABP, hematocrit, serum protein concentration, or red blood cell counts. In this study, Greyhounds with soft, left basilar systolic murmurs had mildly (but significantly) higher mean peak aortic velocities than similar dogs without murmurs. In the dogs with murmurs (and higher velocities), we could not identify structural abnormalities, such as valvular lesions or other congenital defects. There was no inverse correlation between the systolic murmur and the higher hematocrit and red blood cell counts observed in this breed. This 1-2/6 basilar systolic murmur is common in Greyhounds, and it does not appear to be of any clinical consequence.     

Multiplex real-time PCR SYBR Green for detection and typing of group III Clostridium botulinum

Clostridium botulinum type C and type D belonging to the group III organisms, are mainly responsible for animal botulism outbreaks. Clinical signs alone are often insufficient to make a diagnosis of botulism and a laboratory confirmation is required. Laboratory confirmation can be performed by demonstrating the presence of botulinum neurotoxins in serum, gastrointestinal contents, liver, wound of sick or dead animals, or by demonstrating the presence of C. botulinum in gastrointestinal contents, liver, and wound. Demonstration of spores in gastrointestinal contents or tissue of animals with clinical signs indicative of botulism reinforces the clinical diagnosis. With the aim of detecting and typing C. botulinum group III organisms, a multiplex real-time PCR SYBR Green was developed and in-house validated. Selectivity, limit of detection, relative accuracy, relative specificity, relative sensitivity, and repeatability of the method were investigated. The multiplex real-time PCR SYBR green used showed a 100% selectivity, 100% relative accuracy, 100% relative specificity, 100% relative sensitivity and a limit of detection of 277 and 580 DNA copies for C. botulinum type C and C. botulinum type D, respectively. The method reported here represents a suitable tool for laboratory diagnosis of type C and D botulism and for testing a large number of samples collected during the animal botulism surveillance and prevention activities.     

Escherichia coli lipopolysaccharides and Staphylococcus aureus enterotoxin B differentially modulate inflammatory microRNAs in bovine monocytes

MicroRNAs (miRNAs) are a family of regulatory molecules involved in many physiological processes, including activation of cells of the immune system. This study investigated the effect of Escherichia coli lipopolysaccharide (LPS) and Staphylococcus aureus enterotoxin B (SEB) on the expression of five miRNAs involved in the inflammatory response, including miR-9, miR-125 b, miR-155, miR-146 a and miR-223, in bovine CD14(+) cells (monocytes). Incubation of monocytes with SEB induced down-regulation of miR-155, miR-223 and miR-125 b, but not the anti-inflammatory miRNA miR-146 a. Conversely, incubation with LPS upregulated both miR-155 and miR-146 a. In vitro incubation of isolated CD14(+) bovine monocytes with LPS and SEB elicited different and opposite expression of miRNAs reportedly involved in inflammatory reactions.     

Stomatocytosis of Standard Schnauzers is not associated with stomatin deficiency

Stomatocytosis resembles human overhydrated hereditary stomatocytosis (OHSt), a disease characterised by a reduced or absent stomatin expression. The objective of this report was to investigate the expression level of stomatin in erythrocytes from Standard Schnauzers with stomatocytosis. Routine haematology, intraerythrocytic Na(+)/K(+) concentration and stomatin expression were evaluated in blood from twelve Standard Schnauzers and from three controls. SDS-PAGE and Western blotting on isolated integral membrane proteins were used to investigate stomatin expression. Circulating stomatocytes, macrocytosis, anisocytosis, increased erythrocyte fragility and high intracellular sodium and potassium concentrations were found in 10/12 dogs from the same breeding line although stomatin levels were similar to those of controls. In spite of the clinico-pathological similarities between human and canine stomatocytosis, erythrocytes from affected dogs do not lack stomatin and the expression level of this protein cannot therefore be used to diagnose hereditary stomatocytosis in Standard Schnauzers.