Biology

Background: Saccharomyces boulardii is a probiotic yeast routinely used to prevent and to treat gastrointestinal disorders, including the antibiotic-associated diarrhea caused by Clostridium difficile infections. However, only 1-3% of the yeast administered orally is recovered alive in the feces suggesting that this yeast is unable to survive the acidic environment of the gastrointestinal tract.

Inclusive fitness theory predicts that members of non-clonal societies will gain by directing altruistic acts towards their closest relatives. Multiple mating by queens and multiple queens creates distinct full-sister groups in many hymenopteran societies within which nepotism might occur. However, the weight of empirical data suggests that nepotism within full-sister groups is absent. It has been suggested that a lack of reliable recognition markers is responsible. In this paper, we investigated whether epicuticular lipids could provide reliable cues for intracolony kin recognition in two species of social wasps, the paper wasp Polistes dominulus and the hornet Vespa crabro. Epicuticular lipids have previously been shown to be central to kin recognition at the nest level, making them excellent candidates for within-nest discrimination. We genotyped individuals using DNA microsatellites and analysed surface chemistry by gas chromatographymass spectrometry. We find that in both species epicuticular lipids typically could provide enough information to distinguish related nest-mates from unrelated nest-mates, a difference that occurs in colonies with multiple queens. However, in V. crabro, where colonies may be composed by different patrilines, information for discrimination between full sisters and half-sisters is weaker and prone to errors. Our data suggest that epicuticular lipids at best provide reliable information for intracolony nepotistic discrimination in multiple-queen colonies composed of unrelated lines.

The resolution of social conflict in colonies may accord with the interests of the most numerous party. In social insect colonies with single once-mated queens, workers are more closely related to the workers' sons than they are to the queens' sons. Therefore, they should prefer workers to produce males, against the queen's interests. Workers are capable of producing males as they arise from unfertilized eggs. We found Polistes gallicus to have colonies of single, once-mated queens, as determined by microsatellite genotyping of the workers, so worker interests predict worker male production. In colonies lacking queens, workers produced the males, but not in colonies with original queens. Thus worker interests were expressed only when the queen was gone. The high fraction of missing queens and early end to the colony cycle relative to climate so early in the season is surprising and may indicate a forceful elimination of the queen.

Within-colony kin discrimination has not been demonstrated conclusively for any social insect, perhaps partly because highly polymorphic genetic markers necessary to assess within-colony relatednesses have only recently become available. We use microsatellite loci to investigate within-colony kin discrimination in behavioural interactions in the neotropical multiple-queen wasp, Parachartergus colobopterus. Withincolony kin discrimination would be particularly advantageous in this species since average genetic relatedness among colony members overall is low (0.32AE0.06), compared to the relatedness value between full sisters of 0.75. Using seven colonies of individually marked females, we recorded behavioural interactions that were cooperative (222 grooming, 2438 feeding), aggressive (511 body or wing biting, 240 mandible biting) or neutral (1676 antennating). We expected cooperative behaviours to favour closer kin and aggressive behaviours to be directed towards more distant kin, but found that none of the behaviours we investigated showed discrimination on the basis of relatedness. We could have detected a di¡erence in relatedness values of as little as between 0.03 and 0.12, depending on the behaviour being analysed. Thus, we found no evidence for kin discrimination in within-colony behaviour in this species.

Microsatellites have proven to be very useful genetic markers for studies of kinship, parentage, and gene mapping. If microsatellites are conserved among species, then those developed for one species can be used on related species, which would save the time and effort of developing new loci. We evaluated conservation of 27 trinucleotide loci that were derived from 2 species of Polistes wasps in cross-species applications on 27 species chosen from the major lineages of the Vespidae, which diverged as much as 144 million years ago. We further investigated cross-species polymorphism levels for 18 of the loci. There was a clear relationship between cladistic distance and both conservation of the priming sites and heterozygosity. However the loci derived from P. bellicosus were much more widely conserved and polymorphic than were those derived from P. annularis. The disparity in cross-species utility between these sets of loci means that caution should be used in generalizing from conservation rates derived from single species. We found no relationship between locus conservation or heterozygosity and GC content of flanks, repeat motif, repeat length, or heterozygosity in the original species, which suggests that generalizations from other studies reporting such patterns are premature. 1998 Academic Press

Background Social wasps in the subfamily Polistinae (Hymenoptera: Vespidae) have been important in studies of the evolution of sociality, kin selection, and within colony conflicts of interest. These studies have generally been conducted within species, because a resolved phylogeny among species is lacking. We used nuclear DNA microsatellite flanking sequences, mitochondrial COI sequence, and morphological characters to generate a phylogeny for the Polistinae (Hymenoptera) using 69 species. Results Our phylogeny is largely concordant with previous phylogenies at higher levels, and is more resolved at the species level. Our results support the monophyly of the New World subgenera of Polistini, while the Old World subgenera are a paraphyletic group. All genera for which we had more than one exemplar were supported as monophyletic except Polybia which is not resolved, and may be paraphyletic. Conclusion The combination of DNA sequences from flanks of microsatellite repeats with mtCOI sequences and morphological characters proved to be useful characters establishing relationships among the different subgenera and species of the Polistini. This is the first detailed hypothesis for the species of this important group.

Species form the fundamental units of analysis in many areas of biology and, therefore, rigorous delimitation of this unit is important to a broad array of researchers. Recently, many new empirical methods have been proposed to delimit species in nature, and a large literature exists on the theoretical merit and superiority of each method. However, few empirical studies actually compare the results of these methods applied in the same study system. We used a large allozyme and chromosome dataset to apply a number of genetic-distance, character-based, and tree-based methods to a well-studied, data-rich system: the Sceloporus grammicus lizard complex of central Mexico. We hypothesized species boundaries under a general lineage or evolutionary species conceptual framework in an a priori fashion using mapped restriction-site data (mitochondrial DNA and nuclear rDNA), allozymes, and morphology. We then compared the ability of different methods to recover the ''hypothesized evolutionary species'' (HES). Highton's genetic-distance method and a tree-based method consistently recovered all four HES, although sometimes with weak support. With two exceptions, other methods recovered the same HES, but additional groups were weakly delimited and nested within the HES. Given the apparent recent divergence of some of the chromosome races and distinct populations in this complex, these are encouraging results. We emphasize the value of specifying testable criteria as clearly as possible and testing these with methods that make use of different properties of a single dataset.

One of the central questions in physiological ecology is how energetic constraints affect organismal performance and the dynamics of ecological systems. Social insect colonies integrate the balance of supply and demand across levels of biological organization such that the individual components are simultaneously serving as the supply transport network and also the source of energetic demand. An increasing number of studies have demonstrated that the per-capita metabolic rates of individuals within social insect colonies decrease with increasing colony size, a metabolic hypometry much like the pattern exhibited by individual organisms. An important question is thus, whether this scaling pattern is a result of an energetic supply constraint or evidence for an emergent economy of scale. This review synthesizes theoretical models and results from empirical studies on the scaling of resource supply and demand in social insect colonies. Scaling in biology is a powerful tool to unify the study of diverse concepts and organisms; increased integration of mechanistic realism into metabolic models will improve our understanding of the evolution of complex biological systems.

1 Insect metabolic rates are highly variable and are affected by acute environmental and behavioral, developmental, and evolutionary factors.
2 The effects of temperature on insect metabolic rates depend on their behavior, life-history stage, morphology, and size. In many cases, inactive insect metabolic rates increase with temperature in a manner consistent with the assumptions of the metabolic theory of ecology (MTE), but exceptions include insects that are flying, endothermic, or behaviorally thermoregulating. In these cases metabolic rates may remain constant or decrease with increasing temperature.
3 Insect metabolic rates are not generally constrained by oxygen limitation.
4 The metabolic rates for behaviorally active insects may be elevated up to 30 times greater than their standard resting metabolic rates, an aerobic scope greater than the comparable range found among similarly sized vertebrates.
5 Nutritional state can have dramatic influences on insect metabolic rates, ranging from extreme diapause in response to starvation to nearly 10-fold increases in metabolic rate following feeding.
6 Metabolic rate correlates with insect body size both intra- and interspecifically. The interspecific slope is ¾, as predicted by MTE. Individual insects as well as eusocial insect colonies share common hypometric scaling exponents, but there is extensive variation in the metabolic elevation (i.e., scaling intercept or normalization constant) of these allometric relationships. While some of this variation may be related to methodology and behavioral variation, it is likely that these patterns may reflect previously unrecognized evolutionary differences in physiology and life history.
7 Future extensions of MTE should include physiological, behavioral, and evolutionary mechanisms. Future developments of MTE have great potential to investigate a number of areas in which further research is in highly needed including the evolution of insect endothermy, body size, eusociality, and metabolic symmorphosis.

Background: Despite advances in imaging techniques, real-time visualization of the structure and dynamics of tissues and organs inside small living animals has remained elusive. Recently, we have been using synchrotron x-rays to visualize the internal anatomy of millimeter-sized opaque, living animals. This technique takes advantage of partially-coherent x-rays and diffraction to enable clear visualization of internal soft tissue not viewable via conventional absorption radiography. However, because higher quality images require greater x-ray fluxes, there exists an inherent tradeoff between image quality and tissue damage.

Rhythmic tracheal compression is a prominent feature of internal dynamics in multiple orders of insects. During compression parts of the tracheal system collapse, effecting a large change in volume, but the ultimate physiological significance of this phenomenon in gas exchange has not been determined. Possible functions of this mechanism include to convectively transport air within or out of the body, to increase the local pressure within the tracheae, or some combination thereof. To determine whether tracheal compressions are associated with excurrent gas exchange in the ground beetle Pterostichus stygicus, we used flow-through respirometry and synchrotron x-ray phase-contrast imaging to simultaneously record CO(2) emission and observe morphological changes in the major tracheae. Each observed tracheal compression (which occurred at a mean frequency and duration of 15.6+/-4.2 min(-1) and 2.5+/-0.8 s, respectively) was associated with a local peak in CO(2) emission, with the start of each compression occurring simultaneously with the start of the rise in CO(2) emission. No such pulses were observed during inter-compression periods. Most pulses occurred on top of an existing level of CO(2) release, indicating that at least one spiracle was open when compression began. This evidence demonstrates that tracheal compressions convectively pushed air out of the body with each stroke. The volume of CO(2) emitted per pulse was 14+/-4 nl, representing approximately 20% of the average CO(2) emission volume during x-ray irradiation, and 13% prior to it. CO(2) pulses with similar volume, duration and frequency were observed both prior to and after x-ray beam exposure, indicating that rhythmic tracheal compression was not a response to x-ray irradiation per se. This study suggests that intra-tracheal and trans-spiracular convection of air driven by active tracheal compression may be a major component of ventilation for many insects.

Insects in general, and Drosophila in particular, are much more capable of surviving anoxia than vertebrates, and the mechanisms involved are of considerable biomedical and ecological interest. Temperature is likely to strongly affect both the rates of damage occurring in anoxia and the recovery processes in normoxia, but as yet there is no information on the effect of this crucial variable on recovery rates from anoxia in any animal. We studied the effects of temperature, and thus indirectly of metabolic flux rates, on survival and recovery times of individual male Drosophila melanogaster following anoxia and O 2 reperfusion. Individual flies were reared at 25°C and exposed to an anoxic period of 7.5, 25, 42.5 or 60min at 20, 25 or 30°C. Before, during and after anoxic exposure the flies' metabolic rates (MRs), rates of water loss and activity indices were recorded. Temperature strongly affected the MR of the flies, with a Q 10 of 2.21. Temperature did not affect the slope of the relationship between time to recovery and duration of anoxic exposure, suggesting that thermal effects on damage and repair rates were similar. However, the intercept of that relationship was significantly lower (i.e. recovery was most rapid) at 25°C, which was the rearing temperature. When temperatures during exposure to anoxia and during recovery were switched, recovery times matched those predicted from a model in which the accumulation and clearance of metabolic end-products share a similar dependence on temperature.

The negative allometric scaling of metabolic rate with body size is among the most striking patterns in biology. We investigated whether this pattern extends to physically independent eusocial systems by measuring the metabolic rates of whole functioning colonies of the seed-harvester ant Pogonomyrmex californicus. These intraspecific scaling data were compared to the predictions of an additive model developed to estimate collective metabolic rates. Contrary to the prediction of the additive model, colony metabolic rate allometry resembled the pattern commonly observed interspecifically for individual organisms, scaling with colony mass 0.75 . Among the same-aged colonies, net growth rate varied by up to sevenfold, with larger colonies exhibiting higher net growth efficiency than smaller colonies. Isolated worker groups exhibited isometric metabolic rate scaling, suggesting that the social environment of the colony is critical to regulating individual patterns of work output. Within the social environment, individual worker locomotor velocities exhibited power-law distributions that scaled with colony size so that larger colonies exhibited a greater disparity between active and inactive ants than did smaller colonies. These results demonstrate that behavioral organization within colonies may have a major influence on colonylevel metabolism and in generating intraspecific variation in growth trajectories.

Cocaine epigenetically regulates gene expression via changes in histone post-translational modifications (HPTMs). We previously found that the immediate early gene Nr4a1 is epigenetically activated by cocaine in mouse brain reward regions. However, few studies have examined multiple HPTMs at a single gene. Bivalent gene promoters are simultaneously enriched in both activating (H3K4me3 (K4)) and repressive (H3K27me3 (K27)) HPTMs. As such, bivalent genes are lowly expressed but poised for activity-dependent gene regulation. In this study, we identified K4&K27 bivalency at Nr4a1 following investigator-administered cocaine in male and female mice. We applied sequential chromatin immunoprecipitation and qPCR to define Nr4a1 bivalency and expression in striatum (STR), prefrontal cortex (PFC), and hippocampus (HPC). We used Pearson’s correlation to quantify relationships within each brain region across treatment conditions for each sex. In female STR, cocaine increased Nr4a1 mRNA while mai...