Matrix-assisted laser desorption ionization-time of flight mass spectrometry for direct bacterial identification from positive blood culture pellets.

An ammonium chloride erythrocyte-lysing procedure was used to prepare a bacterial pellet from positive blood cultures for direct matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry analysis. Identification was obtained for 78.7% of the pellets tested. Moreover, 99% of the MALDI-TOF identifications were congruent at the species level when considering valid scores. This fast and accurate method is promising.

Effects of corticosterone pellets on baseline and stress-induced corticosterone and corticosteroid-binding-globulin.

Exogenous administration of glucocorticoids is a widely used and efficient tool to investigate the effects of elevated concentrations of these hormones in field studies. Because the effects of corticosterone are dose and duration-dependent, the exact course of plasma corticosterone levels after exogenous administration needs to be known. We tested the performance of self-degradable corticosterone pellets (implanted under the skin) in elevating plasma corticosterone levels. We monitored baseline (sampled within 3min after capture) total corticosterone levels and investigated potential interactions with corticosteroid-binding-globulin (CBG) capacity and the endogenous corticosterone response to handling in Eurasian kestrel Falco tinnunculus and barn owl Tyto alba nestlings. Corticosterone pellets designed for a 7-day-release in rodents elevated circulating baseline total corticosterone during only 2-3 days compared to placebo-nestlings. Highest levels occurred 1-2days after implantation and levels decreased strongly thereafter. CBG capacity was also increased, resulting in a smaller, but still significant, increase in baseline free corticosterone levels. The release of endogenous corticosterone as a response to handling was strong in placebo-nestlings, but absent 2 and 8 days after corticosterone pellet implantation. This indicates a potential shut-down of the hypothalamo-pituitary-adrenal axis after the 2-3 days of elevated baseline corticosterone levels. 20 days after pellet implantation, the endogenous corticosterone response to handling of nestlings implanted with corticosterone pellets attained similar levels as in placebo-nestlings. Self-degradable pellets proved to be an efficient tool to artificially elevate circulating baseline corticosterone especially in field studies, requiring only one intervention. The resulting peak-like elevation of circulating corticosterone, the concomitant elevation of CBG capacity, and the absence of an endogenous corticosterone response to an acute stressor have to be taken into account.

Prophylaxis with resin in wood ants

Animals may use plant compounds to defend themselves against parasites. Wood ants, Formica paralugubris, incorporate pieces of solidified conifer resin into their nests. This behaviour inhibits the growth of bacteria and fungi in nest material and protects the ants against some detrimental microorganisms. Here, we studied the resin-collecting behaviour of ants under field and laboratory conditions. First, we focused on an important assumption of the self-medication hypothesis, which is that the animals deliberately choose the active plant material. In field cafeteria tests, the ants indeed showed a strong preference for resin over twigs and stones, which are building materials commonly encountered in their environment. We detected seasonal variation in the choice of ants: the preference for resin over twigs was more pronounced in spring than in summer, whereas in autumn the ants collected twigs and resin at equal rates. Second, we found almost similar seasonal patterns when comparing the collecting rates of pieces of wood that had been impregnated with turpentine (a distillate of oleoresin) and untreated pieces of wood, which reveals that the preference for resin is based on odour cues. Third, we tested whether the collection of resin is prophylactic or therapeutic. We found that the relative collection rate of resin versus stones did not depend on an experimental infection with the entomopathogenic fungus Metarhizium anisopliae in laboratory colonies. Together, these results show that the ants deliberately choose the resin and suggest that resin collection is prophylactic rather than therapeutic.

Wood ants protect their brood with tree resin

Social insects use multiple lines of collective defences to combat pathogens. One example of a behav- ioural group defence is the use of antimicrobial plant compounds to disinfect the nest. Indeed, wood ants collect coniferous tree resin, and the presence of resin in their nest protects them against fungal and bacterial pathogens. Many questions remain on the mechanisms of resin use, including which factors elicit resin collection and placement within nests. Here, we investigated whether the presence of brood induces Formica paralugubris workers to collect more resin, and whether the workers preferentially place resin near the brood. We also tested whether the collection and placement of resin depends on the presence of the fungal entomopathogen Beauveria bassiana. Workers brought more resin to their nest when brood was present, and preferentially placed the resin near the brood. In contrast, workers did not increase resin collection in response to exposure to B. bassiana, nor did they place resin closer to contaminated brood or contaminated areas of the nest. This lack of response may be explained by a limited effect of resin against the germination and growth of B. bassiana in vitro. Overall, our main result is that woods ants actively position resin near the brood, which probably confers prophylactic protection against other detrimental microorganisms.

Determination of radionuclide levels in rainwater using ion exchange resin and gamma-spectrometry.

The evaluation of radioactivity accidentally released into the atmosphere involves determining the radioactivity levels of rainwater samples. Rainwater scavenges atmospheric airborne radioactivity in such a way that surface contamination can be deduced from rainfall rate and rainwater radioactivity content. For this purpose, rainwater is usually collected in large surface collectors and then measured by gamma-spectrometry after such treatments as evaporation or iron hydroxide precipitation. We found that collectors can be adapted to accept large surface (diameter 47mm) cartridges containing a strongly acidic resin (Dowex AG 88) which is able to quantitatively extract radioactivity from rainwater, even during heavy rainfall. The resin can then be measured by gamma-spectrometry. The detection limit is 0.1Bq per sample of resin (80g) for (137)Cs. Natural (7)Be and (210)Pb can also be measured and the activity ratio of both radionuclides is comparable with those obtained through iron hydroxide precipitation and air filter measurements. Occasionally (22)Na has also been measured above the detection limit. A comparison between the evaporation method and the resin method demonstrated that 2/3 of (7)Be can be lost during the evaporation process. The resin method is simple and highly efficient at extracting radioactivity. Because of these great advantages, we anticipate it could replace former rainwater determination methods. Moreover, it does not necessitate the transportation of large rainwater volumes to the laboratory.

Owl pellets as a source of DNA for genetic studies of small mammals.

Owl pellets contain a good skeletal record of the small mammals consumed, and correspond to the undigested portions of prey which are regurgitated. These pellets are easy to find at the roosting site of owls. As it has been demonstrated that amplifiable DNA can be isolated from ancient bone remains, the possibility of using owl pellets as a source of DNA for small mammal genetics studies via the polymerase chain reaction has been investigated. The main uncertainties when isolating DNA from such a material are firstly the possibility that the extracted DNA would be too degraded during the digestion in the stomach of the owl, and secondly that extensive cross-contaminations could occur among the different prey consumed. The results obtained clearly demonstrate that cross-contamination does not occur, and that mitochondrial and nuclear DNA can be amplified using skulls of small mammals found in owl pellets as a source of DNA. The relative efficiency of two methods of DNA extraction is estimated and discussed. Thus, owl pellets represent a non-invasive sampling technique which provides a valuable source of DNA for studying population genetics of small mammals.

The presence of conifer resin decreases the use of the immune system in wood ants

1. Wood ants (Formica paralugubris) incorporate large amounts of solidified conifer resin into their nest, which reduces the density of many bacteria and fungi and protects the ants against some detrimental micro-organisms. By inducing an environment unfavourable to pathogens, the presence of resin may allow workers to reduce the use of their immune system. 2. The present study tested the hypothesis that the presence of resin decreases the immune activity of wood ants. Specifically, three components of the humoral immune defences of workers kept in resin-rich and resin-free experimental nests (antibacterial, lytic, and prophenoloxidase activities) were compared. 3. The presence of resin was associated with reduced bacterial and fungal densities in nest material and with a small decrease in worker antibacterial and lytic activities. The prophenoloxidase activity was very low in all workers and was not affected by the presence of resin. 4. These results suggest that collective medication with resin reduces pathogen pressure, which in turn decreases the use of the inducible part of the immune system. More generally, the use of plant secondary compounds might be an efficient and economical way to fight pathogens.

Wood ants use resin to protect themselves against pathogens

Social life is generally associated with an increased exposure to pathogens and parasites, due to factors such as high population density, frequent physical contact and the use of perennial nest sites. However, sociality also permits the evolution of new collective behavioural defences. Wood ants, Formica paralugubris, commonly bring back pieces of solidified coniferous resin to their nest. Many birds and a few mammals also incorporate green plant material into their nests. Collecting plant material rich in volatile compounds might be an efficient way to fight bacteria and fungi. However, no study has demonstrated that this behaviour has a positive effect on survival. Here, we provide the first experimental evidence that animals using plant compounds with antibacterial and antifungal properties survive better when exposed to detrimental micro-organisms. The presence of resin strongly improves the survival of F. paralugubris adults and larvae exposed to the bacteria Pseudomonas fluorescens, and the survival of larvae exposed to the entomopathogenic fungus Metarhizium anisopliae. These results show that wood ants capitalize on the chemical defences which have evolved in plants to collectively protect themselves against pathogens.