The global impact of Wolbachia on mitochondrial diversity and evolution.

The spread of maternally inherited microorganisms, such as Wolbachia bacteria, can induce indirect selective sweeps on host mitochondria, to which they are linked within the cytoplasm. The resulting reduction in effective population size might lead to smaller mitochondrial diversity and reduced efficiency of natural selection. While documented in several host species, it is currently unclear if such a scenario is common enough to globally impact the diversity and evolution of mitochondria in Wolbachia-infected lineages. Here, we address this question using a mapping of Wolbachia acquisition/extinction events on a large mitochondrial DNA tree, including over 1000 species. Our analyses indicate that on a large phylogenetic scale, other sources of variation, such as mutation rates, tend to hide the effects of Wolbachia. However, paired comparisons between closely related infected and uninfected taxa reveal that Wolbachia is associated with a twofold reduction in silent mitochondrial polymorphism, and a 13% increase in nonsynonymous substitution rates. These findings validate the conjecture that the widespread distribution of Wolbachia infections throughout arthropods impacts the effective population size of mitochondria. These effects might in part explain the disconnection between genetic diversity and demographic population size in mitochondria, and also fuel red-queen-like cytonuclear co-evolution through the fixation of deleterious mitochondrial alleles.

Evaluation of the transarticular external skeletal fixator for the treatment of tarsocrural instability in 32 cats.

The medical records of all cats with tarsocrural joint instability that were treated between June 2002 and December 2008 at the Royal Veterinary College were retrospectively reviewed. A total of 32 cats were identified. Information gathered included signalment, type of injury (subluxation or luxation), concurrent fractures, presence of soft tissue wounds, transarticular external skeletal fixation (TESF) type, configuration of TESF (number of pins proximal and distal to the joint), duration of hospitalisation, duration of TESF prior to removal, complications and cost. A significant association was identified between the length of hospitalisation and the presence of wounds. Similarly a significant association was present between wounds and final cost of treatment. Additionally, the authors found that a high number of implant related complications were present when only two pins were used proximal and distal to the tarsocrural joint, but this association was not significant.

Inter-island divergence within Drosophila mauritiana, a species of the D. simulans complex: Past history and/or speciation in progress?

Speciation with gene flow may be more common than generally thought, which makes detailed understanding of the extent and pattern of genetic divergence between geographically isolated populations useful. Species of the Drosophila simulans complex provide a good model for speciation and evolutionary studies, and hence understanding their population genetic structure will increase our understanding of the context in which speciation has occurred. Here, we describe genetic diversity and genetic differentiation of two distant populations of D. mauritiana (Mauritius and Rodrigues Islands) at mitochondrial and nuclear loci. We surveyed the two populations for their mitochondrial haplotypes, eight nuclear genes and 18 microsatellite loci. A new mitochondrial type is fixed in the Rodrigues population of D. mauritiana. The two populations are highly differentiated, their divergence appears relatively ancient (100,000 years) compared to the origin of the species, around 0.25MYA, and they exhibit very limited gene flow. However, they have similar levels of divergence from their sibling, D. simulans. Both nuclear genes and microsatellites revealed contrasting demographic histories between the two populations, expansion for the Mauritius population and stable population size for the Rodrigues Island population. The discovery of pronounced geographic structure within D. mauritiana combined to genetic structuring and low gene flow between the two island populations illuminates the evolutionary history of the species and clearly merits further attention in the broad context of speciation.

Spontaneous gastroduodenal perforations in dogs: a retrospective study of 15 cases.

The clinical findings and outcomes following surgical management of spontaneous gastroduodenal perforations in 15 dogs were reviewed in a retrospective study to identify related risk factors. Clinical and clinicopathological findings were diverse. Use of multiple diagnostic procedures yielded a strong index of suspicion for gastrointestinal perforation. There was a trend towards improved survival for animals with perforation of the gastric fundus/body compared to pyloric or duodenal perforation, although the difference was not statistically significant. Five dogs were euthanased at surgery; two dogs died within two days after surgery. Seven of the eight surviving dogs had a favourable long-term outcome. Previous administration of NSAIDs was the only identified predisposing factor in ten of 10 of the dogs.

Plasma lactate concentrations and blood gas values in dogs undergoing surgical attenuation of a single congenital portosystemic shunt.

Plasma concentration of lactate and the values of pH, pO(2) and pCO(2) were measured in the portal, systemic venous and, when possible, systemic arterial blood of 31 dogs with a single congenital portosystemic shunt, before and shortly after the temporary complete occlusion of the shunt, and at the end of surgery. At completion of the surgery, the shunt in 16 of the dogs had been occluded completely whereas in the other 15 it had been occluded only partially. There were no significant differences between any of the measurements of these variables in the portal venous, systemic venous or arterial plasma of any of the dogs, or between the values measured in the groups in which the shunts had been occluded completely or partially. Furthermore, there were no significant differences between the two groups of dogs in the arteriovenous gradients calculated at any of the sampling sites or sampling times. None of the variables was associated with the development of postoperative complications.

Cauda equina syndrome treated by lumbosacral stabilisation in a cat.

In this case report, a case of cauda equina syndrome arising from lumbosacral disease in an eight-year-old Burmese cat is described. The cat had a history of chronic right pelvic limb lameness. Neurological evaluation was consistent with a lesion involving the cauda equina. Radiographic findings were consistent with a diagnosis of lumbosacral disease. Due to perceived dorsoventral lumbosacral instability, dorsal decompression and stabilisation of the lumbosacral junction was performed. An original method of spinal stabilisation for this indication is described. The cat had a successful long-term outcome with complete resolution of its presenting clinical signs.

Investigation of task performance variations according to task requirements and alertness across the 24-h day in shift workers.

The study investigated time-of-day effects on task performance in shift workers in different tasks (reaction time, discrimination, probe recognition, free recall), by varying task-specific features. On each of six recordings, each programmed on a different day and in a randomised order, operators rated alertness and performed different tasks. Self-rated alertness varied according to a typical diurnal trend. Time of day also affected reaction time (slower responses at 03:00 hours), discrimination performance (lower accuracy at 03:00 hours in the most difficult condition) and recall (superior recall at 07:00 and 11:00 hours following deeper processing at encoding). The data demonstrated time-of-day effects on cognitive processes also involved in many real-job activities, despite the lack of control for a number of exogenous factors known to interfere with performance in work settings. Since in the cognitively more loaded tasks, time-of-day effects depended on task conditions, the findings are of operational concern in shift-work situations involving differential task requirements. In a real-job setting, performance variations were observed according to time of day and task requirements in a set of cognitively more or less demanding tasks. Task-specific research across the 24-h day enables a better understanding of operators' tasks and the development of supporting technology.

Inclusion of marbofloxacin in PMMA orthopaedic cement: an in vitro experimental study.

This in vitro experimental study investigated the feasibility for marbofloxacin, a veterinary fluoroquinolone antibiotic, to retain its antibacterial activity after its inclusion in polymethylmethacrylate (PMMA) cement. The assays were conducted on gelose cultures of various types of bacteria isolated from canine spontaneous osteomyelitis. The efficiency of the antibiotics was assessed by using an antibiogram method. Resistance of marbofloxacin to the temperature observed during PMMA polymerization, antimicrobial effect of galenic, useful concentrations and comparison with gentamicin (reference antibiotic for this use) were evaluated. Marbofloxacin retained its antimicrobial activity after being heated to high temperatures reached during polymerization. The more effective galenic form to incorporate into the PMMA monomer was the marbofloxacin powder and the appropriate concentration was 1/40 degrees . In this experiment, marbofloxacin included in PMMA showed a good antimicrobial activity; however this activity was lower than gentamicin added to PMMA on Gram + and Pseudomonas bacteria.Therefore, it seems useful to incorporate marbofloxacin to PMMA cement to treat, or to prevent, osteomyelitis associated with marbofloxacin sensitive bacteria. Nevertheless, the development of a marbofloxacin-PMMA cement requires further evaluation, especially pharmacological, biomechanical and clinical.

Electrolytic electrospray ionization mass spectrometry of C(60)-TTF-C(60) derivatives: high-resolution mass measurement and molecular ion gas-phase reactivity.

The electrochemical process involved in electrospray ionization is used to obtain odd-electron molecular ions from C(60)-TTF-C(60) and its methano derivatives. Exact mass measurements obtained using high-resolution mass spectrometry are reported, and the gas-phase behavior of the radical cation (retro-Diels-Alder reaction and [M/2 + H](+) ion formation) is described.

Divergence between Drosophila santomea and allopatric or sympatric populations of D. yakuba using paralogous amylase genes and migration scenarios along the Cameroon volcanic line.

We have used two paralogous genes (Amyrel and Amy) of the amylase multigene family to reconstruct the phylogeny of the nine Drosophila melanogaster subgroup sister species, including D. santomea, the newly discovered endemic from São Tomé island. The evolutionary divergence of these genes is of special interest as it is suspected to result from physiological evolution via gene duplication. This paper describes the relationship between the geographical origin of the various strains and the patterns of mating and phylogeny, focusing on the evolution of D. santomea and its relationship to other species and their niches. The Amyrel and Amy data indicate that, contrary to expectations, the sympatric insular D. yakuba population is less closely related to D. santomea than allopatric mainland ones, suggesting that the extant insular D. yakuba population on São Tomé results from a recent secondary colonization. Data for sympatric and allopatric D. yakuba suggest that D. santomea arose from a mainland D. yakuba parental stock when montane habitats of the Cameroon volcanic line extended to lower altitudes during colder and less humid periods. Despite their different modes of evolution and different functions, the Amyrel and Amy genes provide remarkably consistent topologies and hence reflect the same history, that of the species.

Morphological and molecular differentiation of invasive freshwater species of the genus Corbicula (Bivalvia, corbiculidea) suggest the presence of three taxa in French rivers.

Asiatic Clams are common in brackish and fresh water in Asia, and they were introduced into North America in 1924 and have now spread throughout the continent. During the last two decades they have been reported in Europe, but the number of species here is uncertain. Populations of Corbicula from France and the Netherlands were analysed morphologically and genetically to quantify the degree of species and/or population differentiation. The morphological and genetic data, based on allozymes and mitochondrial sequences, were in full agreement. They indicate that there are two distinct species, identified as C. fluminalis and C. fluminea, in the two countries. Analyses of the mitochondrial COI gene revealed an unexpected divergent population of Corbicula in the Rhône. All these individuals were morphologically identified as C. fluminea, but had a COI sequence different from the two previous species. This population may, therefore, be a more ancient population, or a distinct species introduced via a different colonization route.

Molecular characterization and evolution of the amylase multigene family of Drosophila ananassae.

Drosophila ananassae is known to produce numerous alpha-amylase variants. We have cloned seven different Amy genes in an African strain homozygous for the AMY1,2,3,4 electrophoretic pattern. These genes are organized as two main clusters: the first one contains three intronless copies on the 2L chromosome arm, two of which are tandemly arranged. The other cluster, on the 3L arm, contains two intron-bearing copies. The amylase variants AMY1 and AMY2 have been assigned to the intronless cluster, and AMY3 and AMY4 to the second one. The divergence of coding sequences between clusters is moderate (6.1% in amino acids), but the flanking regions are very different, which could explain their differential regulation. Within each cluster, coding and noncoding regions are conserved. Two very divergent genes were also cloned, both on chromosome 3L, but very distant from each other and from the other genes. One is the Amyrel homologous (41% divergent), the second one, Amyc1 (21.6% divergent) is unknown outside the D. ananassae subgroup. These two genes have unknown functions.

Evolutionary novelties in islands: Drosophila santomea, a new melanogaster sister species from São Tomé.

The finding of new melanogaster sister species may help us in understanding more about how the emergence of genetic novelties, particularly in insular habitats, can result in speciation. Here we report on the discovery of Drosophila santomea, which is the first melanogaster sibling found off West-equatorial Africa, on São Tomé, one of the Gulf of Guinea islands. Although the eight other melanogaster sister species are remarkably conservative in their morphology except for their terminalia, the new find has a morphological trait distinguishing it from all of these: a pure yellow body coloration of both sexes without the normal black abdominal banding. Evidence from the terminalia, polytene and mitotic chromosomes, period gene and allozymes are provided indicating that it is nonetheless the nearest relative of Drosophila yakuba with which it coexists on the island. The new find is a clear-cut taxon as shown by the production of sterile male hybrids, eventually with developmental defects, in both directions of cross with yakuba and by the existence of an altitudinal divide accompanied by a hybrid zone at mid-elevation on the island. Molecular and karyotypic data further support this conclusion. In contrast to the significant divergence of their nuclear DNAs, an intriguing similarity in their cytochrome b sequences was observed indicating a recent coalescence common to santomea, yakuba and also teissieri cytoplasms. These were shown to harbour the same Wolbachia endosymbiotic bacteria which could possibly be responsible for mitochondrial DNA hitchhiking across the species barrier.

Density and variability of dinucleotide microsatellites in the parthenogenetic polyploid snail Melanoides tuberculata.

Characterization of microsatellites in the parthenogenetic polyploid snail Melanoides tuberculata revealed an unusual high density of dinucleotide repeats. Multiple banding patterns were obtained at these loci, and interpreted as a consequence of polyploidy. Microsatellite variability was low within, but high between, shell morphotypes. Genotypes were wholly transmitted from mothers to offspring. These results suggest that reproduction is strictly apomictic, and that shell morphotypes are genetic clones.

Metabolic evolution in alpha-amylases from Drosophila virilis and D. repleta, two species with different ecological niches.

alpha-Amylases from Drosophila virilis and D. repleta were partially purified by ion exchange chromatography. The two amylases share common characteristics for pH and cations effects, although with slight differences. D. virilis has optimal activity at pH 6.6 and D. repleta at pH 7.2. Calcium, sodium, and potassium cations activate amylolytic activity in both species but Ba2+ has an activation effect in D. repleta only. In contrast, there are major differences in thermal offbility and kinetics among amylases of the two species. D. virilis amylase is much more stable at high temperature and the optimal temperatures are very different between the two species, respectively, 45 degrees C and 30 degrees C for D. virilis and D. repleta. alpha-Amylase activity using different substrates is greater on starch than on glycogen in both species and still higher on amylose for D. virilis, the nonfungus feeder species. alpha-Amylase of D. repleta, the mycophagous species, has a better affinity to amylopectin and glycogen. Such differences in substrate specificity suggest adaptation to different resources in these species living in different habitats. Metabolic evolution seems to have occurred through a "tradeoff" between kinetic effectiveness and the nature of substrate, with a higher Vmax on amylose for D. virilis and a lower K(m) on glycogen for D. repleta.

Amyrel, a paralogous gene of the amylase gene family in Drosophila melanogaster and the Sophophora subgenus.

We describe a gene from Drosophila melanogaster related to the alpha-amylase gene Amy. This gene, which exists as a single copy, was named Amyrel. It is strikingly divergent from Amy because the amino acid divergence is 40%. The coding sequence is interrupted by a short intron at position 655, which is unusual in amylase genes. Amyrel has also been cloned in Drosophila ananassae, Drosophila pseudoobscura, and Drosophila subobscura and is likely to be present throughout the Sophophora subgenus, but, to our knowledge, it has not been detected outside. Unexpectedly, there is a strong conservation of 5' and 3' flanking regions between Amyrel genes from different species, which is not the case for Amy and which suggests that selection acts on these regions. In contrast to the Amy genes, Amyrel is transcribed in larvae of D. melanogaster but not in adults. However, the protein has not been detected yet. Amyrel evolves about twice as fast as Amy in the several species studied. We suggest that this gene could result from a duplication of Amy followed by accelerated and selected divergence toward a new adaptation.

Distribution and evolution of introns in Drosophila amylase genes.

While the two amylase genes of Drosophila melanogaster are intronless, the three genes of D. pseudoobscura harbor a short intron. This raises the question of the common structure of the Amy gene in Drosophila species. We have investigated the presence or absence of an intron in the amylase genes of 150 species of Drosophilids. Using polymerase chain reaction (PCR), we have amplified a region that surrounds the intron site reported in D. pseudoobscura and a few other species. The results revealed that most species contain an intron, with a variable size ranging from 50 to 750 bp, although the very majoritary size was around 60-80 bp. Several species belonging to different lineages were found to lack an intron. This loss of intervening sequence was likely due to evolutionarily independent and rather frequent events. Some other species had both types of genes: In the obscura group, and to a lesser extent in the ananassae subgroup, intronless copies had much diverged from intron-containing genes. Base composition of short introns was found to be variable and correlated with that of the surrounding exons, whereas long introns were all A-T rich. We have extended our study to non-Drosophilid insects. In species from other orders of Holometaboles, Lepidoptera and Hymenoptera, an intron was found at an identical position in the Amy gene, suggesting that the intron was ancestral.

Variation in sex-, stage- and tissue-specific expression of the amylase genes in Drosophila ananassae.

Expression of the amylase multigene family of Drosophila ananassae was investigated in third-instar larvae and adults. A developmental differentiation was found between the Amy1-2 and Amy3-4 gene clusters, the former being preferentially expressed in larvae, the latter in adults. During adult life, we observed a decrease in Amy1-2 expression in males of certain strans. We have raised some arguments for the existence of trans-active regulators, acting as repressors of Amy1-2 in adults. The putative repressors might exhibit a geographical polymorphism, with a fixed active form in Pacific regions and a polymorphic pattern in Africa, thus increasing the diversity observed in adult amylase phenotypes. A clear differentiation between the two gene clusters was also found in tissue-specific activity along the third-instar larval midgut. In the anterior midgut, only Amy1-2 is active, while both gene groups are expressed in the posterior midgut, with an additional subzonation within it.

Evolution of the response patterns to dietary carbohydrates and the developmental differentiation of gene expression of alpha-amylase in Drosophila.

Intraspecific variation of alpha-amylase activity in D. melanogaster and D. immigrans, which is distantly related to D. melanogaster, and interspecific variation of alpha-amylase activity in 18 Drosophila species were examined. The amount of intraspecific variation of alpha-amylase activities measured in terms of coefficient of variation in D. melanogaster and D. immigrans was one-half and one-tenth or less, respectively, of the interspecific variation in 18 Drosophila species. We also surveyed the response patterns of alpha-amylase activity to dietary carbohydrates at the larval and adult stages. The levels of alpha-amylase activity depended on both repression by dietary glucose (glucose repression) and induction by dietary starch (starch induction). In general, our data suggest that glucose repression was conserved among species at both stages while starch induction was mainly observed in larvae, although the degree of the response depended on species. In D. lebanonensis lebanonensis and D. serrata, larvae expressed electrophoretically different alpha-amylase variants (isozymes) from those of adult flies. These results may suggest that the regulatory systems responsible both for the response to environment and developmental expression are different among species in Drosophila.