Aggregation and species coexistence in fleas parasitic on small mammals

The aggregation model of coexistence states that species coexistence is facilitated if interspecific aggregation is reduced relative to intraspecific aggregation. We investigated the relationship between intraspecific and interspecific aggregation in 17 component communities (the flea assemblage of a host population) of fleas parasitic on small mammals and hypothesized that interspecific interactions should be reduced relative to intraspecific interactions, facilitating species coexistence. We predicted that the reduction of the level of interspecific aggregation in relation to the level of intraspecific aggregation would be positively correlated with total flea abundance and species richness of flea assemblages. We also expected that the higher degree of facilitation of flea coexistence would be affected by host parameters such as body mass, basal metabolic rate (BMR) and depth and complexity of burrows. Results of this study supported the aggregation model of coexistence and demonstrated that, in general, a) conspecific fleas were aggregated across their hosts; b) flea assemblages were not dominated by negative interspecific interactions; and c) the level of interspecific aggregation in flea assemblages was reduced in relation to the level of intraspecific aggregation. Intraspecific aggregation tended to be correlated positively to body mass, burrow complexity and mass-independent BMR of a host. Positive interspecific associations of fleas tended to occur more frequently in species-rich flea assemblages and/or in larger hosts possessing deep complex burrows. Intraspecific aggregation increased relative to interspecific aggregation when species richness of flea infracommunities (the flea assemblage of a host individual) and component communities increased. We conclude that the pattern of flea coexistence is related both to the structure of flea communities and affinities of host species.     

Assembly rules of ectoparasite communities across scales: combining patterns of abiotic factors,host composition,geographic space,phylogeny and traits

We investigated the role of environmental filtering as an underlying mechanism of assembly of compound communities of fleas parasitic on Palearctic small mammals at two spatial scales; a continental scale (encompassing regions across the entire Palearctic) and a regional scale (across sampling localities within Slovakia). We used the three‐table ordination (the RLQ analysis) and its extended version that links species occurrences with geographic space, environmental variables, and species traits and phylogeny (the ESLTP analysis). We asked whether environmental filtering acts as an assembly rule of compound communities of fleas and, if yes, a) whether the effect of environment on species composition of compound communities of fleas differs between spatial scales and b) what are the relative importance of the abiotic and host environments. We found that compound communities of fleas are, to a great extent, assembled via environmental filters that represent interplay between filtering via abiotic environment and filtering via host composition. The relative importance of these two components of environmental filtering differed between spatial scales. Host composition had a stronger effect on flea assembly than abiotic environment on the continental scale, while the opposite was true for the regional scale. The likely reason behind this scale‐dependence is that communities on the regional scale are mainly governed by ecological and epidemiological processes, while communities on the continental scale are mainly affected by evolutionary, biogeographic and historical forces.     

Co‐occurrence and phylogenetic distance in communities of mammalian ectoparasites: limiting similarity versus environmental filtering

Similarity between species plays a key role in the processes governing community assembly. The co‐occurrence of highly similar species may be unlikely if their similar needs lead to intense competition (limiting similarity). On the other hand, persistence in a particular habitat may require certain traits, such that communities end up consisting of species sharing the same traits (environmental filtering). Relatively little information exists on the relative importance of these processes in structuring parasite communities. Assuming that phylogenetic relatedness reflects ecological similarity, we tested whether the co‐occurrence of pairs of flea species (Siphonaptera) on the same host individuals was explained by the phylogenetic distance between them, among 40 different samples of mammalian hosts (rodents and shrews) from different species, areas or seasons. Our results indicate that frequency of co‐occurrence between flea species increased with decreasing phylogenetic distance between them in 37 out of 40 community samples, with 14 of these correlations being statistically significant. A meta‐analysis across all samples confirmed the overall trend for closely related species to co‐occur more frequently on the same individual hosts than expected by chance, independently of the identity of the host species or of environmental conditions. These findings suggest that competition between closely related, and therefore presumably ecologically similar, species is not important in shaping flea communities. Instead, if only fleas with certain behavioural, ecological and physiological properties can encounter and exploit a given host, and if phylogenetic relationships determine trait similarity among flea species, then a process akin to environmental filtering, or host filtering, could favour the co‐occurrence of related species on the same host.     

Host Specificity, Parasite Community Size and the Relation between Abundance and its Variance

We investigated the empirical relationship between mean abundance and its variance, known as Taylor’s power law, in fleas parasitic on small mammals. It has been suggested that the exponent of this function, b, represents a true biological character of a species and, dependent on the level of host specificity, varies among species. Other empirical and theoretical studies suggest that exponent b depends on interspecific competition and varies intraspecifically. We tested these hypotheses using data from central and eastern Slovakia. We demonstrate that the slope of Taylor’s relationship (a) is repeatable within a flea species, i.e. the slope represents a true species character; (b) increases with an increase of the degree of flea host specificity; and (c) decreases with an increase in flea community size. We discuss our results with the idea that the host can mediate interactions among and within flea species. Co-ordinating editor: A. Biere     

Intracellular calcium measurements as a method in studies on activity of purinergic P2X receptor channels

Extracellular nucleotide-activated purinergic receptors (P2XRs) are a family of cation-permeable channels that conduct small cations, including Ca²⁺, leading to the depolarization of cells and subsequent stimulation of voltage-gated Ca²⁺ influx in excitable cells. Here, we studied the spatiotemporal characteristics of intracellular Ca²⁺ signaling and its dependence on current signaling in excitable mouse immortalized gonadotropin-releasing hormone-secreting cells (GT1) and nonexcitable human embryonic kidney cells (HEK-293) cells expressing wild-type and chimeric P2XRs. In both cell types, P2XR generated depolarizing currents during the sustained ATP stimulation, which desensitized in order (from rapidly desensitizing to nondesensitizing): P2X₃R > P2X_2b + X₄R > P2X_2bR > P2X_2a + X₄R > P2X₄R > P2X_2aR > P2X₇R. HEK-293 cells were not suitable for studies on P2XR-mediated Ca²⁺ influx because of the coactivation of endogenously expressed Ca²⁺-mobilizing purinergic P2Y receptors. However, when expressed in GT1 cells, all wild-type and chimeric P2XRs responded to agonist binding with global Ca²⁺ signals, which desensitized in the same order as current signals but in a significantly slower manner. The global distribution of Ca²⁺ signals was present independently of the rate of current desensitization. The temporal characteristics of Ca²⁺ signals were not affected by voltage-gated Ca²⁺ influx and removal of extracellular sodium. Ca²⁺ signals reflected well the receptor-specific EC₅₀ values for ATP and the extracellular Zn²⁺ and pH sensitivities of P2XRs. These results indicate that intracellular Ca²⁺ measurements are useful for characterizing the pharmacological properties and messenger functions of P2XRs, as well as the kinetics of channel activity, when the host cells do not express other members of purinergic receptors. ATP-gated receptor channels; inward currents; intracellular calcium signals; desensitization-inactivation; voltage-gated calcium influx; localized and global calcium signals     

Endothelin-induced, long lasting, and Ca2+ influx-independent blockade of intrinsic secretion in pituitary cells by Gz subunits

The G protein-coupled receptors in excitable cells have prominent roles in controlling Ca2+-triggered secretion by modulating voltage-gated Ca2+ influx. In pituitary lactotrophs, spontaneous voltage-gated Ca2+ influx is sufficient to maintain prolactin release high. Here we show that endothelin in picomolar concentrations can interrupt such release for several hours downstream of spontaneous and high K+-stimulated voltage-gated Ca2+ influx. This action occurred through the Gz signaling pathway; the adenylyl cyclase-signaling cascade could mediate sustained inhibition of secretion, whereas rapid inhibition also occurred at elevated cAMP levels regardless of the status of phospholipase C, tyrosine kinases, and protein kinase C. In a nanomolar concentration range, endothelin also inhibited voltage-gated Ca2+ influx through the G i/o signaling pathway. Thus, the coupling of seven-transmembrane domain endothelin receptors to Gz proteins provided a pathway that effectively blocked hormone secretion distal to Ca2+ entry, whereas the cross-coupling to G i/o proteins reinforced such inhibition by simultaneously reducing the pacemaking activity.     

Structural and gene expression analyses of uptake hydrogenases and other proteins involved in nitrogenase protection in Frankia

The actinorhizal bacterium Frankia expresses nitrogenase and can therefore convert molecular nitrogen into ammonia and the by-product hydrogen. However, nitrogenase is inhibited by oxygen. Consequently, Frankia and its actinorhizal hosts have developed various mechanisms for excluding oxygen from their nitrogen-containing compartments. These include the expression of oxygen-scavenging uptake hydrogenases, the formation of hopanoid-rich vesicles, enclosed by multi-layered hopanoid structures, the lignification of hyphal cell walls, and the production of haemoglobins in the symbiotic nodule. In this work, we analysed the expression and structure of the so-called uptake hydrogenase (Hup), which catalyses the in vivo dissociation of hydrogen to recycle the energy locked up in this ‘waste’ product. Two uptake hydrogenase syntons have been identified in Frankia: synton 1 is expressed under free-living conditions while synton 2 is expressed during symbiosis. We used qPCR to determine synton 1 hup gene expression in two Frankia strains under aerobic and anaerobic conditions. We also predicted the 3D structures of the Hup protein subunits based on multiple sequence alignments and remote homology modelling. Finally, we performed BLAST searches of genome and protein databases to identify genes that may contribute to the protection of nitrogenase against oxygen in the two Frankia strains. Our results show that in Frankia strain ACN14a, the expression patterns of the large (HupL1) and small (HupS1) uptake hydrogenase subunits depend on the abundance of oxygen in the external environment. Structural models of the membrane-bound hydrogenase subunits of ACN14a showed that both subunits resemble the structures of known [NiFe] hydrogenases (Volbeda et al. 1995), but contain fewer cysteine residues than the uptake hydrogenase of the Frankia DC12 and Eu1c strains. Moreover, we show that all of the investigated Frankia strains have two squalene hopane cyclase genes (shc1 and shc2). The only exceptions were CcI3 and the symbiont of Datisca glomerata, which possess shc1 but not shc2. Four truncated haemoglobin genes were identified in Frankia ACN14a and Eu1f, three in CcI3, two in EANpec1 and one in the Datisca glomerata symbiont (Dg).