Ice crystallization by Pseudomonas syringae

Several bacterial species can serve as biological ice nuclei. The best characterized of these is Pseudomonas syringae, a widely distributed bacterial epiphyte of plants. These biological ice nuclei find various applications in different fields, but an optimized production method was required in order to obtain the highly active cells which may be exploited as ice nucleators. The results presented here show that P. syringae cells reduce supercooling of liquid or solid media and enhance ice crystal formation at sub-zero temperatures, thus leading to a remarkable control of the crystallization phenomenon and a potential for energy savings. Our discussion focuses on recent and future applications of these ice nucleators in freezing operations, spray-ice technology and biotechnological processes. Received: 21 December 1999 / Received revision: 29 February 2000 / Accepted: 6 March 2000     

Die Leistungsersparnis beim Verbandsflug

Zusammenfassung Für einen Tragflügel endlicher Spannweite werden die Entstehung der Auftriebskraft und das Auftreten eines Aufwindfeldes in der Umgebung des Flügels beschrieben. Beim Verbandsflug fliegt jeder Flügel in dem Aufwind aller übrigen, wodurch sich eine beträchtliche Leistungsersparnis ergibt. Es wird über ein theoretisches Verfahren berichtet, mit dem diese Leistungsersparnis für beliebig gestaltete Flugformationen mit beliebiger Zahl der beteiligten Vögel berechnet werden kann. Bei diesem Verfahren werden die schlagenden Flügel durch starre Flügel ersetzt. Es wird gezeigt, daß dies eine sinnvolle Vereinfachung darstellt.Die theoretischen Rechnungen ergaben, daß die Leistungsersparnis des gesamten Verbandes nur von der Gesamtzahl der Vögel und von deren seitlichem Abstand abhängt. Die Verteilung dieser Leistungsersparnis auf die beteiligten Individuen hingegen ist von der Form des Verbandes abhängig. Es werden Ergebnisse von Beispielrechnungen mitgeteilt, bei denen die Verteilung der Leistungsersparnis auf die einzelnen Vögel in symmetrischen und unsymmetrischen Winkelflugformationen einschließlich der Schräglinien-Anordnung sowie auch in ganz beliebig geformten Verbänden untersucht wurde. Dabei zeigte sich, daß in Winkelformationen mit geradlinigen Schenkeln das an der Spitze des Verbandes fliegende Individuum die geringste Leistungsersparnis aufzuweisen hat. Durch systematische Veränderungen der Form des Verbandes wurde eine Formation mit nahezu gleichförmiger Verteilung der Leistungsersparnis auf die beteiligten Vögel gefunden. Verschiedene Erscheinungen des Verbandsfluges von Vögeln werden aus aerodynamischer Sicht diskutiert.

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The power reduction in flight formations

Summary For a wing of finite span the generation of lift and the occurrence of an induced upwash field in the vicinity of the wing are described. In formation flight each wing is situated in an upwash field generated by all other wings of the formation, leading to a considerable reduction in flight power demand. A theoretical method is presented to calculate the flight power reduction for arbitrarily shaped flight formations with any number of birds. In this method, the beating wings are replaced by fixed wings, which is shown to be an adequate simplification.Theoretical calculations show, that the total flight power reduction of the whole formation depends only on the total number of birds and on their lateral distance. The distribution of this power reduction on the individuals involved, however, is a function of the actual shape of the formation. Results of numerical examples are presented, which show the distribution of power reduction on the individuals involved in symmetrical and unsymmetrical vee formations including the flight in a straight line as well as in arbitrarily shaped formations. In vee formations with straight sides, the individual at the apex of the formation has the smallest reduction in flight power demand. By systematic modifications of the shape a formation with almost uniform distribution of flight power reduction was found. Some aspects of formation flight of birds are discussed from an aerodynamicist's point of view.

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Vortrag gehalten auf der 84. Jahresversammlung der DO-G zu Saarbrücken.     

The function of flight formations in Greylag Geese Anser anser; energy saving or orientation?

Twenty five formations of Greylag Geese Anser anser were photographed from immediately below to eliminate perspective distortion, and the lateral and longitudinal displacements of the birds relative to each other were measured. We scaled the photographs and used measurements of bill to tail and wing span made on 15 freshly shot birds to convert the lateral displacements to wing-tip spacings. The birds flew on average with an overlap of their wing tips (median =17 cm) which corresponded very closely with the overlap expected (16 cm) from an aerodynamic model which predicted the position which maximized energy savings. However, the variation in positions was large, and only 17% of birds actually flew in the optimum range. The mean saving in induced power averaged across the distribution of positions was 26.5%, and the contribution to total flight costs was a reduction of 4.5–9%. This saving was greater than we found previously in the Pink-footed Goose Anser brachyrhynchus. There was no correlation between position in the skein and longitudinal displacement, as would be anticipated if the birds were equalizing the savings across the skein. This does not mean costs were not equalized because other mechanisms are possible. There was no correlation between depth and wing tip spacing which does not support the orientation/communication hypothesis. Body size and thus flight costs may be a factor influencing the function of formation flights.     

Impaired Control of Body Cooling during Heterothermia Represents the Major Energetic Constraint in an Aging Non-Human Primate Exposed to Cold

Daily heterothermia is used by small mammals for energy and water savings, and seems to be preferentially exhibited during winter rather than during summer. This feature induces a trade-off between the energy saved during daily heterothermia and the energy cost of arousal, which can impact energy balance and survival under harsh environmental conditions. Especially, aging may significantly affect such trade off during cold-induced energy stress, but direct evidences are still lacking. We hypothesized that aging could alter the energetics of daily heterothermia, and that the effects could differ according to season. In the gray mouse lemur (Microcebus murinus), a non-human primate species which exhibits daily heterothermia, we investigated the effects of exposures to 25 and 12°C on body composition, energy balance, patterns of heterothermia and water turnover in adult (N = 8) and aged animals (N = 7) acclimated to winter-like or summer-like photoperiods.Acclimation to summer prevented animals from deep heterothermia, even during aging. During winter, adult animals at 12°C and aged animals at 25°C exhibited low levels of energy expenditure with minor modulations of heterothermia. The major effects of cold were observed during winter, and were particularly pronounced in aged mouse lemurs which exhibited deep heterothermia phases. Body composition was not significantly affected by age and could not explain the age-related differences in heterothermia patterns. However, aging was associated with increased levels of energy expenditure during cold exposure, in concomitance with impaired energy balance. Interestingly, increased energy expenditure and depth of heterothermia phases were strongly correlated.In conclusion, it appeared that the exhibition of shallow heterothermia allowed energy savings during winter in adult animals only. Aged animals exhibited deep heterothermia and increased levels of energy expenditure, impairing energy balance. Thus, an impaired control of the heterothermic process induced high energy costs in the aging mouse lemur exposed to cold.     

Conodont biostratigraphy of Tournaisian shallow-water carbonates in central Guangxi,South China

The conodont faunas of Tournaisian shallow-water carbonates from central Guangxi are described mainly for biostratigraphic purposes. A complete series of samples was collected from the Long’an and Du’an formations in the Long’an section. These formations are characterized by lime-mudstone, skeletal and peloidal wackestone, packstone and grainstone with typical shallow-water biota. Overall, these samples produced 809 identifiable Pl elements, belonging to 50 species in 11 genera, of which one species and one subspecies are new. The fauna enables the establishment of seven biozones (in ascending order): Polygnathus spicatus, Siphonodella homosimplex, S. sinensis, S. dasaibaensis, Polygnathus communis carina Acme, Gnathodus cuneiformis, and P. communis porcatus zones. Based on these new collections from central Guangxi and on data from the literature, a conflated Tournaisian conodont zonation is proposed for shallow-water successions in South China. Most of the conodont zones correlate well with their counterparts recognized in Western Europe, which may be of greater significance in stratigraphic correlation than previously thought.     

Biochemical content, energy composition and reproductive effort in the broadcasting sea star Asterias vulgaris over the spawning period

To assess sex differences in reproductive effort, we examined the biochemical composition and energetic content of the principal body components of the broadcast spawning sea star Asterias vulgaris in the Mingan Islands in the northern Gulf of St. Lawrence, eastern Canada. The body wall was the most stable body component, showing no variations in mass or in lipid and protein content (and total energetic content) between sexes or during spawning. Patterns in the gonads differed between sexes and with spawning. The lipid, protein and carbohydrate content of the ovary dropped during spawning, while only the protein content of the testis decreased significantly. Reproductive effort, expressed as loss of energy in the gonads during spawning for an individual weighing 10 g in underwater mass (8.2 cm in radius), was six times greater in females (49.5 kJ) than males (7.9 kJ). The energetic content of the pyloric caeca also decreased during spawning, by 17.7 kJ in females and 21.5 kJ in males, mainly due to a decrease in lipids. If this decrease is included as reproductive effort, it lessens the gender difference. The caecum decrease possibly represented expenditures due to formation of aggregations or the expulsion of gametes during spawning. Effectively, we observed aggregations during a massive spawning in this population. The sex ratio did not differ from 1:1 in all size classes sampled. This suggests that, unless males suffer higher mortality, females manage to allocate as much energy to somatic growth as males, possibly by feeding at higher rates to compensate for their higher reproductive effort. Stomach protein content tended to be higher in females than males and may indicate greater muscular development to facilitate digestion.     

Comparison of natural and pharmacological hypothermia in animals: Determination of activation energy of metabolism

The constancy of the activation energy of metabolism (E) for all living organisms is one of the most impressive, though controversial, statements of the modern metabolic theory of evolution. According to WBE-theory suggested by West, Brown, and Enquist, E should be in the range from −0.6 to −0.7 eV. However, there are many examples of significant deviations of E from the predictions of the theory. Now we have conducted a study of this value using rats in different types of pharmacological hypothermia: 1. Short-term (for several hours) hypothermia induced by anesthetic xylazine; 2. Daily torpor-like state induced by the pharmacological composition developed in our previous study. It has been found that in pharmacological daily hypothermia E = −0.56 ± 0.03 eV, which was close to that in daily heterotherms found in literature, E = −0.57 ± 0.04 eV. In short-term hypothermia E was substantially lower, E = −0.17 ± 0.071 eV. Our analysis revealed that in short-term hypothermia, changes in body temperature may lag behind changes in metabolic rate for a period Δt, affecting E. We propose an approach for estimating Δt and obtaining an adjusted E = −0.68 ± 0.17 eV, which corresponds to theoretical predictions. We assume that a similar consideration of Δt should be done when calculating E of daily heterotherms. We assume that in ectotherms, when the ambient temperature changes rapidly, changes in metabolic rate may lag behind changes in body temperature for a period (−) Δt, that should also be considered in E calculations. The proposed approach may contribute to the further development of the metabolic theory of evolution and may be useful in comparing artificial and natural hypothermia, as well as in studying the energy transformations in ecosystems.     

An analytic treatment of the condensation of phospholipid chains

A theory relating hydrocarbon chain length to temperature is developed and applied to the liquid crystalline to crystalline phase transiion in phospholipids. Interactions in both the chains and head groups are considered. The hydrocarbon tails are treated as freely jointed chains and the heads as a system of interacting dipoles. Analytic expressions are obtained for the internal energy, entropy, free energy and specific heat of the combined system. The free energy is minimized to obtain a relationship between straightening force and chain length. Agreement is good with experiments involving variation of water content and % cholesterol.Work was supported by a grant from the National Research Council of Canada.     

Modular construction for function of a ribonucleoprotein enzyme: the catalytic domain of Bacillus subtilis RNase P complexed with B. subtilis RNase P protein

The bacterial RNase P holoenzyme catalyzes the formation of the mature 5′-end of tRNAs and is composed of an RNA and a protein subunit. Among the two folding domains of the RNase P RNA, the catalytic domain (C-domain) contains the active site of this ribozyme. We investigated specific binding of the Bacillus subtilis C-domain with the B.subtilis RNase P protein and examined the catalytic activity of this C-domain–P protein complex. The C-domain forms a specific complex with the P protein with a binding constant of ~0.1 µM. The C-domain–P protein complex and the holoenzyme are equally efficient in cleaving single-stranded RNA (~0.9 min^–1 at pH 7.8) and substrates with a hairpin–loop 3′ to the cleavage site (~40 min^–1). The holoenzyme reaction is much more efficient with a pre-tRNA substrate, binding at least 100-fold better and cleaving 10–500 times more efficiently. These results demonstrate that the RNase P holoenzyme is functionally constructed in three parts. The catalytic domain alone contains the active site, but has little specificity and affinity for most substrates. The specificity and affinity for the substrate is generated by either the specificity domain of RNase P RNA binding to a T stem–loop-like hairpin or RNase P protein binding to a single-stranded RNA. This modular construction may be exploited to obtain RNase P-based ribonucleoprotein complexes with altered substrate specificity.     

Variable domain antibodies specific for viral hemorrhagic septicemia virus (VHSV) selected from a randomized IgNAR phage display library

Phage display libraries are used to screen for nucleotide sequences that encode immunoglobulin variable (V) regions that are specific for a target antigen. We previously constructed an immunoglobulin new antigen receptor (IgNAR) phage display library. Here we used this library to obtain an IgNAR V region that is specific for viral hemorrhagic septicemia virus (VHSV). A phage clone (clone 653) was found to be specific for VHSV by the biopanning method. The V region of clone 653 was used to construct a 6 × His tagged recombinant IgNAR-653 V protein (rIgNAR-653) using the Escherichia coli pET system. The rIgNAR-653 protein bound specifically to VHSV, confirming its activity.     

Experimental and Theoretical Bases of Specific Affinity, a Cytoarchitecture-Based Formulation of Nutrient Collection Proposed To Supercede the Michaelis-Menten Paradigm of Microbial Kinetics

A theory for solute uptake by whole cells was derived with a focus on the ability of oligobacteria to sequester nutrients. It provided a general relationship that was used to obtain the kinetic constants for in situ marine populations in the presence of naturally occurring substrates. In situ affinities of 0.9 to 400 liters g of cells⁻¹ h⁻¹ found were up to 10³ times smaller than those from a “Marinobacter arcticus ” isolate, but springtime values were greatly increased by warming. Affinities of the isolate for usual polar substrates but not for hydrocarbons were diminished by ionophores. A kinetic curve or Monod plot was constructed from the best available data for cytoarchitectural components of the isolate by using the theory together with concepts and calculations from first principles. The order of effect of these components on specific affinity was membrane potential > cytoplasmic enzyme concentration > cytoplasmic enzyme affinity > permease concentration > area of the permease site > translation coefficient > porin concentration. Component balance was influential as well; a small increase in cytoplasmic enzyme concentration gave a large increase in the effect of permease concentration. The effect of permease concentration on specific affinity was large, while the effect on K_m was small. These results are in contrast to the Michaelis-Menten theory as applied by Monod that has uptake kinetics dependent on the quality of the permease molecules, with K_m as an independent measure of affinity. Calculations demonstrated that most oligobacteria in the environment must use multiple substrates simultaneously to attain sufficient energy and material for growth, a requirement consistent with communities largely comprising few species.     

Oral Ondansetron Administration in Emergency Departments to Children with Gastroenteritis: An Economic Analysis

Background

The use of antiemetics for children with vomiting is one of the most controversial decisions in the treatment of gastroenteritis in developed countries. Ondansetron, a selective serotonin receptor antagonist, has been found to be effective in improving the success of oral rehydration therapy. However, North American and European clinical practice guidelines continue to recommend against its use, stating that evidence of cost savings would be required to support ondansetron administration. Thus, an economic analysis of the emergency department administration of ondansetron was conducted. The primary objective was to conduct a cost analysis of the routine administration of ondansetron in both the United States and Canada.

Methods and Findings

A cost analysis evaluated oral ondansetron administration to children presenting to emergency departments with vomiting and dehydration secondary to gastroenteritis from a societal and health care payer''s perspective in both the US and Canada. A decision tree was developed that incorporated the frequency of vomiting, intravenous insertion, hospitalization, and emergency department revisits. Estimates of the monetary costs associated with ondansetron use, intravenous rehydration, and hospitalization were derived from administrative databases or emergency department use. The economic burden in children administered ondansetron plus oral rehydration therapy was compared to those not administered ondansetron employing deterministic and probabilistic simulations. We estimated the costs or savings to society and health care payers associated with the routine administration of ondansetron. Sensitivity analyses considered variations in costs, treatment effects, and exchange rates. In the US the administration of ondansetron to eligible children would prevent approximately 29,246 intravenous insertions and 7,220 hospitalizations annually. At the current average wholesale price, its routine administration to eligible children would annually save society US$65.6 million (US$49.1–US$81.1) and health care payers US$61.1 million (US$46.2–US$76.3). In Canada the administration of ondansetron to eligible children would prevent 4,065 intravenous insertions and 1,003 hospitalizations annually. Its routine administration would annually save society CDN$1.72 million (CDN$1.15–CDN$1.89) and the health care system CDN$1.18 million (CDN$0.88–CDN$1.41).

Conclusions

In countries where intravenous rehydration is often employed, the emergency department administration of oral ondansetron to children with dehydration and vomiting secondary to gastroenteritis results in significant monetary savings compared to a no-ondansetron policy. Please see later in the article for the Editors'' Summary     

Intramolecular formation and cleavage of C-S/N bonds promoted by iron-sulfur coordination chemistry

The reaction of a tetradentate ligand, 2,2′-(pyridin-2-ylmethylazanediyl)diethanethiol (H₂L), with dodecacarbonyltriiron in dry toluene leads to the formations of a hexacarbonyl complex, [Fe₂(EDT)(CO)₆] (EDT = ethylenedithiolate), 1a, which is fully characterised. The formation of this complex is via intramolecular C-S/N bonds formation/cleavage promoted by iron-sulfur coordination chemistry. A possible mechanism for the C-S/N bonds formation/cleavage in the formation of the complex is proposed.     

Light-Induced Proton Gradient Formation in Intact Cells of Dunaliella salina

A light-induced proton gradient (ΔpH) increase as exhibited by an increase of 9-aminoacridine fluorescence quenching is demonstrated between the external medium and the interior of the halophytic green alga Dunaliella salina. The formation and maintenance of the ΔpH is sensitive to electron transport inhibitors and to uncouplers. It is inhibited by p-chloromercuribenzenesulfonic acid (50% inhibition at 3 micromolar), which does not affect photosynthetic O₂ evolution. It is concluded that the observed ΔpH is located across the plasmalemma or the chloroplast envelope. The formation and maintenance of the light-induced proton gradient requires the presence of Na⁺. Substitution of NaCl by KCl or glycerol results in inhibition of the ΔpH formation. The proton gradient is also sensitive to ATPase and energy transfer inhibitors. It is suggested that a Na⁺/H⁺ pump mechanism may be involved in the formation of the proton gradient in intact Dunaliella cells.     

Kinetic Analysis of Bifidobacterial Metabolism Reveals a Minor Role for Succinic Acid in the Regeneration of NAD+ through Its Growth-Associated Production

Several strains belonging to the genus Bifidobacterium were tested to determine their abilities to produce succinic acid. Bifidobacterium longum strain BB536 and Bifidobacterium animalis subsp. lactis strain Bb 12 were kinetically analyzed in detail using in vitro fermentations to obtain more insight into the metabolism and production of succinic acid by bifidobacteria. Changes in end product formation in strains of Bifidobacterium could be related to the specific rate of sugar consumption. When the specific sugar consumption rate increased, relatively more lactic acid and less acetic acid, formic acid, and ethanol were produced, and vice versa. All Bifidobacterium strains tested produced small amounts of succinic acid; the concentrations were not more than a few millimolar. Succinic acid production was found to be associated with growth and stopped when the energy source was depleted. The production of succinic acid contributed to regeneration of a small part of the NAD⁺, in addition to the regeneration through the production of lactic acid and ethanol.     

Strain energy in the femoral neck during exercise

Physical activity is recommended to mitigate the incidence of hip osteoporotic fractures by improving femoral neck strength. However, results from clinical studies are highly variable and unclear about the effects of physical activity on femoral neck strength. We ranked physical activities recommended for promoting bone health based on calculations of strain energy in the femoral neck. According to adaptive bone-remodeling theory, bone formation occurs when the strain energy (S) exceeds its homeostatic value by 75%. The potential effectiveness of activity type was assessed by normalizing strain energy by the applied external load. Tensile strain provided an indication of bone fracture. External force and joint motion data for 15 low- and high-load weight-bearing and resistance-based activities were used. High-load activities included weight-bearing activities generating a ground force above 1 body-weight and maximal resistance exercises about the hip and the knee. Calculations of femoral loads were based on musculoskeletal and finite-element models. Eight of the fifteen activities were likely to trigger bone formation, with isokinetic hip extension (ΔS=722%), one-legged long jump (ΔS=572%), and isokinetic knee flexion (ΔS=418%) inducing the highest strain energy increase. Knee flexion induced approximately ten times the normalized strain energy induced by hip adduction. Strain and strain energy were strongly correlated with the hip-joint reaction force (R²=0.90–0.99; p<0.05) for all activities, though the peak load location was activity-dependent. None of the exercises was likely to cause fracture. Femoral neck mechanics is activity-dependent and maximum isokinetic hip-extension and knee-flexion exercises are possible alternative solutions to impact activities for improving femoral neck strength.     

Looking for high energy density compounds among polynitraminecubanes

Based on fully optimized geometric structures at DFT-B3LYP/6-311G** level, we calculated electronic structures, heats of formation, strain energies, bond dissociation energies and detonation performance (detonation velocity and detonation pressure) for a series of polynitraminecubanes. Our results have shown that energy gaps of cubane derivatives are much higher than that of triaminotrinitrobenzene (TATB), which means that cubane derivatives may be more sensitive than TATB. Polynitraminecubanes have high and positive heats of formation, and a good linear relationship between heats of formation and nitramine group numbers was presented. As the number of nitramine groups in the molecule increases, the enthalpies of combustion values are increasingly negative, but the specific enthalpy of combustion values decreases. It is found that all cubane derivatives have high strain energies, which are affected by the number and position of nitramine group. The calculated bond dissociation energies of C-NHNO₂ and C-C bond show that the C-C bond should be the trigger bond in the pyrolysis process. It is found that detonation velocity (D), detonation pressure (P) and molecule density (ρ) have good linear relationship with substituented group numbers. Heptanitraminecubane and octanitraminecubane have good detonation performance over 1,3,5,7-tetranitro-1,3,5,7-tetraazacyclooctane (HMX), and they can be regarded as potential candidates of high energy density compounds (HEDCs). The results have not only shown that these compounds may be used as HEDCs, but also provide some useful information for further investigation.     

Occurrence of the earliest known Sphenothallus Hall in the Lower Cambrian of Southern Shaanxi Province, China

Three-dimensionally preserved specimens of an indeterminate species of Sphenothallus Hall occur in limestones in the uppermost Guojiaba and lower Xiannudong formations (Qiongzhusian and early Canglangpuan stages, Lower Cambrian) in southern Shaanxi Province, China. The specimens consist of a small (up to 3 mm long), gradually tapered, subcircular to elliptic (transversely) tube that is broken at both ends, commonly with the fracture edge at the wide (apertural) end of each face forming a V- or U-shaped notch. Some specimens preserve the subconical upper (proximal) part of the holdfast, and all specimens are black. Combined scanning and transmission electron microscopy and energy dispersive elemental analyses show that the tube wall is finely laminated, with individual laminae measuring about 1 μm thick, and consists mainly of calcium phosphate but also contains organic material. Sphenothallus specimens from the Lower Cambrian Guojiaba and Xiannudong formations represent the earliest known record of this genus, and indicate that the origin of this taxon is coeval with and possibly related to the Early Cambrian bioradiation event.     

Biostratigraphie d’après les foraminifères et paléoenvironnements des séries post-Éocène du sondage ASHTART 28, golfe de Gabès (Tunisie)

The Cenozoic sequence of Ashtart 28 well drilled in the Gulf of Gabes (Tunisia) is the subject of a biostratigraphical study. The samples recovered in cuttings from 390 m and downwards allowed to recognize, above the Late Eocene sediments, a sedimentary series, lithologically diversified, nearly 1600 m thick. Marine Pliocene deposits, generally attesting a low bathymetry, lie unconformably above the Messinian (Oued Bel Khedim formation), which shows the usual features of the Mediterranean confinement. The underlying Messinian pre-evaporitic platform series (Melqart formation), that is over 250 m thick, is typical of a perireefal environment. The sediments assigned to the Tortonian (Somâa Sands formation) are continental and occur unconformably above the approximately 500-metres-thick Middle Miocene strata (Saouaf, Mahmoud, Aïn Grab and Salammbô pars formations). The marine Lower Miocene and Oligocene sediments (Salammbô pars and Ketatna formations), that are more than 300 m thick, lie in continuity under the Middle Miocene. The infralittoral Chattian sequence has especially supplied a diversified assemblage of larger foraminifera recovered in other west-mediterranean basins. Datings were obtained based on planktonic and larger benthic foraminifera (Miogypsinidae, Nummulitidae, Lepidocyclinidae) and by correlations obtained by means of well loggings and lithostratigraphy. Benthic foraminifera, mainly listed for the Miocene and Oligocene, are studied from a systematic, stratigraphic and paleogeographic point of view. The paleoenvironments of deposits are defined for each considered stratigraphic interval. Comparisons are sketched with other drillings of the Gulf of Gabes. Thanks to the numerous data obtained by this detailed study, the Ashtart drilling can serve as a reference for the Tertiary sequence of this part of the Mediterranean domain.     

A Remarkable Case of Micro-Endemism in Laonastes aenigmamus (Diatomyidae,Rodentia) Revealed by Nuclear and Mitochondrial DNA Sequence Data

L. aenigmamus is endemic to the limestone formations of the Khammuan Province (Lao PDR), and is strongly specialized ecologically. From the survey of 137 individuals collected from 38 localities, we studied the phylogeography of this species using one mitochondrial (Cyt b) and two nuclear genes (BFIBR and GHR). Cyt b analyses reveal a strong mtDNA phylogeographical structure: 8 major geographical clades differing by 5–14% sequence divergence were identified, most of them corresponding to distinct karst areas. Nuclear markers display congruent results but with a less genetic structuring. Together, the data strongly suggest an inland insular model for Laonastes population structure. With 8 to 16 evolutionary significant units in a small area (about 200×50 km) this represents an exceptional example of micro-endemism. Our results suggest that L. aenigmamus may represent a complex of species and/or sub-species. The common ancestor of all Laonastes may have been widely distributed within the limestone formations of the Khammuan Province at the end of Miocene/beginning of the Pliocene. Parallel events of karst fragmentation and population isolation would have occurred during the Pleistocene or/and the end of the Pliocene. The limited gene flow detected between populations from different karst blocks restrains the likelihood of survival of Laonastes. This work increases the necessity for a strict protection of this rare animal and its habitat and provides exclusive information, essential to the organization of its protection.