Symbiotic streptomycetes in antennal glands of the South American digger wasp genus Trachypus (Hymenoptera,Crabronidae)

Several insect taxa have evolved symbioses with actinobacteria that protect the host or its nutritional resources against pathogens. Digger wasps of the genus Philanthus (‘beewolves’; Hymenoptera, Crabronidae) cultivate ‘Candidatus Streptomyces philanthi’ (Ca. S. philanthi) in specialized antennal glands and transfer them to subterranean brood cells, where the symbionts provide protection for the beewolf larva against pathogens by producing a cocktail of antibiotic substances. The present study investigates the occurrence of antennal symbionts in two species of the genus Trachypus, which is the closest relative to Philanthus. Cross sections of the antennae from females of both Trachypus denticollis and Trachypus boharti reveal bacteria‐containing gland reservoirs that are morphologically very similar to those found in Philanthus spp. Polymerase chain reaction‐based screens with specific primers, sequencing of the partial 16S rRNA gene, and fluorescence in situ hybridization confirm the presence of close relatives of ‘Ca. S. philanthi’ in T. denticollis and T. boharti. However, 16S rRNA sequence divergence between Trachypus spp. and Philanthus spp. symbionts is much higher for T. boharti than for T. denticollis, suggesting that horizontal transmission and/or de novo uptake of symbionts from the environment occurs occasionally. The results obtained indicate that the protective symbiosis with antennal actinobacteria is older and more widespread than previously recognized and occurs in at least two genera of digger wasps that comprise approximately 170 species.     

A cladistic analysis of the subfamily Philanthinae (Hymenoptera: Sphecidae)

Abstract. The Philanthinae, as treated here, contains the eight valid genera Aphilanthops Patton, Cerceris Latreille, Clypeadon Patton, Eucerceris Cresson, Philanthinus Beaumont, Philanthus Fabricius, Pseudoscolia Radoszkowski and Trachypus Klug. Listropygia Bohart is synonymized with Clypeadon , syn.n. The genera Eremiasphecium Kohl and Odontosphex Arnold are removed from the Philanthinae and regarded as genera incertae sedis within Sphecidae. Analysis of the genera of this redefined subfamily Philanthinae finds two major clades whose monophyly is strongly supported: ( Philanthinus + ( Philanthus + Trachypus )) [= Tribe Philanthini] and ( Pseudoscolia + ( Cerceris + Eucerceris )) [= Tribe Cercerini]. There is some evidence for the monophyly of Clypeadon + Aphilanthops [= Tribe Aphilanthopini]. Cerceris and Philanthus are inferred to be paraphyletic with respect to Eucerceris and Trachypus , respectively. Philanthinus is removed from the tribe Aphilanthopini and placed in the tribe Philanthini, and the monotypic tribe Pseudoscoliini is synonymized with Cercerini.     

Morphology and ultrastructure of a bacteria cultivation organ: the antennal glands of female European beewolves, Philanthus triangulum (Hymenoptera, Crabronidae)

Females of a solitary digger wasp, the European beewolf (Philanthus triangulum F.), cultivate symbiotic bacteria of the genus Streptomyces in specialized antennal glands. The streptomycetes are secreted in the subterranean brood cells and protect the offspring against mould fungi. We reconstructed the complex morphology of the antennal glands using 3D-visualization software, investigated the ultrastructure of the glands, and examine the role of the antennal glands as organs for the cultivation of the symbiotic bacteria. The bacteria are cultivated in five antennomeres within large reservoirs that consist of two slightly bent lobes. Each gland reservoir is bordered by a monolayered epithelium lined with a partially reinforced cuticle and when completely filled with bacteria it comprises about half of the antennomere's volume. The opening of the reservoir is covered by gelatinous appendage of the cuticle. The cells of the monolayered epithelium bordering each reservoir show basal invaginations, apical microvilli and numerous vesicles. Each reservoir is surrounded by approximately 400 class 3 gland units that are connected to the reservoir lumen through conducting canals. The class 3 gland cells contain numerous vesicles and a high density of rough endoplasmatic reticulum. In the reservoir lumen, large numbers of symbiotic Streptomyces bacteria are embedded in secretion droplets. Thus, the bacteria are apparently provided with large amounts of nutrients via the gland epithelium and the class 3 gland cell units.     

Life cycle and population dynamics of a protective insect symbiont reveal severe bottlenecks during vertical transmission

Insects engage in mutualistic relationships with a wide variety of microorganisms that are usually transmitted vertically to the next generation. During transmission, the symbiont populations often suffer significant bottlenecks that may entail major genetic and genomic consequences. Here we investigated the life-cycle and the severity of transmission bottlenecks in a symbiotic system with an unusual way of post-hatch vertical transmission by using quantitative PCRs and morphological 3D-reconstructions. European beewolves (Philanthus triangulum, Hymenoptera: Crabronidae) harbor symbiotic bacteria (‘Candidatus Streptomyces philanthi’) in specialized antennal gland reservoirs and secrete them into their subterranean brood cells. The symbionts are later taken up by the beewolf larva and incorporated into the cocoon material to provide protection against pathogenic microorganisms. Even after months of hibernation, the symbiont population on the cocoon is estimated to encompass around 1.4 × 10⁵ cells. However, our results indicate that only few of these bacterial cells (about 9.7 × 10²) are taken up from the cocoon by the emerging female. The symbiont population subsequently undergoes logistic growth within the antennal gland reservoirs and reaches a maximum of about 1.5 × 10⁷ cells 3–4 days after emergence. The maximum specific growth rate is estimated to be 0.084–0.105 h⁻¹. With a total reduction in cell numbers of about 6.7 × 10⁻⁵ during vertical transmission, the symbiont population experiences one of the most severe bottlenecks known for any symbiotic system to date. This extreme bottleneck may have significantly affected the evolution of the beewolf-Streptomyces symbiosis by increased genetic drift, an accumulation of mildly deleterious mutations and genome erosion.     

Symbiotic bacteria protect wasp larvae from fungal infestation

Symbiotic associations between different organisms are of great importance for evolutionary and ecological processes [1-4]. Bacteria are particularly valuable symbiotic partners owing to their huge diversity of biochemical pathways that may open entirely new ecological niches for higher organisms [1-3]. Here, we report on a unique association between a new Streptomyces species and a solitary hunting wasp, the European beewolf (Philanthus triangulum, Hymenoptera, Crabronidae). Beewolf females cultivate the Streptomyces bacteria in specialized antennal glands and apply them to the brood cell prior to oviposition. The bacteria are taken up by the larva and occur on the walls of the cocoon. Bioassays indicate that the streptomycetes protect the cocoon from fungal infestation and significantly enhance the survival probability of the larva, possibly by producing antibiotics. Behavioral observations strongly suggest a vertical transmission of the bacteria. Two congeneric beewolf species harbor closely related streptomycetes in their antennae, indicating that the association with protective bacteria is widespread among philanthine wasps and might play an important role in other insects as well. This is the first report on the cultivation of bacteria in insect antennae and the first case of a symbiosis involving bacteria of the important antibiotic-producing genus Streptomyces.     

Structure, chemical composition and putative function of the postpharyngeal gland of the emerald cockroach wasp, Ampulex compressa (Hymenoptera, Ampulicidae)

The postpharyngeal gland (PPG) plays a major role in the social integration of ant colonies. It had been thought to be restricted to ants but was recently also described for a solitary wasp, the European beewolf (Philanthus triangulum). This finding posed the question whether the gland has evolved independently in the two taxa or has been inherited from a common ancestor and is hence homologous. The latter alternative would be supported if a PPG was found in more basal taxa. Therefore, we examined a species at the base of the Apoidea, the solitary ampulicid wasp Ampulex compressa, for the existence of a PPG. Both sexes of this species possess a cephalic gland that branches off the posterior part of the pharynx, is lined by a cuticular intima and surrounded by a monolayered epithelium with the epithelial cells bearing long hairs. Most of these morphological characteristics conform to those of the PPG of ants and beewolves. Chemical analysis of the gland content revealed that it contains mainly hydrocarbons and that there is a congruence of the pattern of hydrocarbons in the gland, on the cuticle, and in the hemolymph, as has also been reported for both ants and beewolves. Based on these morphological and chemical results we propose that the newly described cephalic gland is a PPG and discuss its possible function in A. compressa. The present study supports the view of a homologous origin of the PPG in the aculeate Hymenoptera.     

Escape from parasitism: spatial and temporal strategies of a sphecid wasp against a specialised cuckoo wasp

Parasites and parasitoids exert an important selection pressure on organisms and, thus, play an important role for both population dynamics and evolutionary responses of host species. We investigated host-parasite interactions in a brood-caring wasp, the European beewolf, Philanthus triangulum (Hymenoptera, Sphecidae), and asked whether females of this species might employ temporal or spatial strategies to reduce the rate of attack by a specialised brood parasitoid, the cuckoo wasp Hedychrum rutilans (Hymenoptera, Chrysididae). Females of the host species might shift their activity to periods of low parasitoid activity both in the course of the season and in the course of the day. On a spatial scale, aggregated or dispersed nesting might be favoured depending on the form of the density dependence of parasitism. The beginning and end of the flight season of host and parasitoid were nearly identical. Activity of chrysidids relative to beewolves did not change significantly during the flight season. However, relative parasitoid activity declined in the course of the day, suggesting the existence of temporal enemy-free space in the evening hours. Shifting the main activity to the evening hours might be a flexible response of beewolves to the presence of chrysidids. Activity of cuckoo wasps per nest was independent of nest density but the actual rate of parasitism as revealed by nest excavations indicated direct density dependence. Total mortality, however, was inversely density dependent. Thus, in the study population aggregated nesting did not reduce parasitism but minimised total mortality.     

Comparative analysis of septic injury-inducible genes in phylogenetically distant model organisms of regeneration and stem cell research, the planarian Schmidtea mediterranea and the cnidarian Hydra vulgaris

Background

Host-parasite interactions are among the most important biotic relationships. Host species should evolve mechanisms to detect their enemies and employ appropriate counterstrategies. Parasites, in turn, should evolve mechanisms to evade detection and thus maximize their success. Females of the European beewolf (Philanthus triangulum, Hymenoptera, Crabronidae) hunt exclusively honeybee workers as food for their progeny. The brood cells containing the paralyzed bees are severely threatened by a highly specialized cuckoo wasp (Hedychrum rutilans, Hymenoptera, Chrysididae). Female cuckoo wasps enter beewolf nests to oviposit on paralyzed bees that are temporarily couched in the nest burrow. The cuckoo wasp larva kills the beewolf larva and feeds on it and the bees. Here, we investigated whether H. rutilans evades detection by its host. Since chemical senses are most important in the dark nest, we hypothesized that the cuckoo wasp might employ chemical camouflage.

Results

Field observations suggest that cuckoo wasps are attacked by beewolves in front of their nest, most probably after being recognized visually. In contrast, beewolves seem not to detect signs of the presence of these parasitoids neither when these had visited the nest nor when directly encountered in the dark nest burrow. In a recognition bioassay in observation cages, beewolf females responded significantly less frequently to filter paper discs treated with a cuticular extract from H. rutilans females, than to filter paper discs treated with an extract from another cuckoo wasp species (Chrysis viridula). The behavior to paper discs treated with a cuticular extract from H. rutilans females did not differ significantly from the behavior towards filter paper discs treated with the solvent only. We hypothesized that cuckoo wasps either mimic the chemistry of their beewolf host or their host's prey. We tested this hypothesis using GC-MS analyses of the cuticles of male and female beewolves, cuckoo wasps, and honeybee workers. Cuticle extracts of Hedychrum nobile (Hymenoptera: Chrysididae) and Cerceris arenaria (Hymenoptera: Crabronidae) were used as outgroups. There was little congruence with regard to cuticular compounds between H. rutilans females and honeybees as well as females of C. arenaria and H. nobile. However, there was a considerable similarity between beewolf females and H. rutilans females. Beewolf females show a striking dimorphism regarding their cuticular hydrocarbons with one morph having (Z)-9-C25:1 and the other morph having (Z)-9-C27:1 as the major component. H. rutilans females were more similar to the morph having (Z)-9-C27:1 as the main component.

Conclusion

We conclude that H. rutilans females closely mimic the composition of cuticular compounds of their host species P. triangulum. The occurrence of isomeric forms of certain compounds on the cuticles of the cuckoo wasps but their absence on beewolf females suggests that cuckoo wasps synthesize the cuticular compounds rather than sequester them from their host. Thus, the behavioral data and the chemical analysis provide evidence that a specialized cuckoo wasp exhibits chemical mimicry of the odor of its host. This probably allows the cuckoo wasp to enter the nest with a reduced risk of being detected by olfaction and without leaving traitorous chemical traces.     

Diversity of endophytic bacteria within nodules of the Sphaerophysa salsula in different regions of Loess Plateau in China

A total of 115 endophytic bacteria were isolated from root nodules of the wild legume Sphaerophysa salsula grown in two ecological regions of Loess Plateau in China. The genetic diversity and phylogeny of the strains were revealed by restriction fragment length polymorphism and sequencing of 16S rRNA gene and enterobacterial repetitive intergenic consensus-PCR. Their symbiotic capacity was checked by nodulation tests and analysis of nifH gene sequence. This is the first systematic study on endophytic bacteria associated with S. salsula root nodules. Fifty of the strains found were symbiotic bacteria belonging to eight putative species in the genera Mesorhizobium, Rhizobium and Sinorhizobium, harboring similar nifH genes; Mesorhizobium gobiense was the main group and 65 strains were nonsymbiotic bacteria related to 17 species in the genera Paracoccus, Sphingomonas, Inquilinus, Pseudomonas, Serratia, Mycobacterium, Nocardia, Streptomyces, Paenibacillus, Brevibacillus, Staphylococcus, Lysinibacillus and Bacillus, which were universally coexistent with symbiotic bacteria in the nodules. Differing from other similar studies, the present study is the first time that symbiotic and nonsymbiotic bacteria have been simultaneously isolated from the same root nodules, offering the possibility to accurately reveal the correlation between these two kinds of bacteria. These results provide valuable information about the interactions among the symbiotic bacteria, nonsymbiotic bacteria and their habitats.     

A ‘social’ gland in a solitary wasp? The postpharyngeal gland of female European beewolves (Hymenoptera,Crabronidae)

Exocrine glands play an important role in maintaining the integrity of colonies of social Hymenoptera. The postpharyngeal gland (PPG) of ants is crucial for the generation of a nest odour that enables nestmate recognition. The evolutionary history of this gland is unknown and it was thought to be restricted to ants. Here we describe an exocrine head gland in females of a solitary crabronid wasp, the European beewolf, Philanthus triangulum, that resembles the PPG of ants in many respects. The newly described gland has the same location and the same glove like shape as in ants, and it also has a monolayered epithelium with similar ultrastructure. Unlike in ants, the epithelium bears hairs that reach into the lumen of the gland. Although the PPG of beewolves serves a completely different function it is also associated to an allogrooming behaviour as in ants. Based on these morphological and behavioural similarities as well as similarities in the chemical composition of the content of the PPG of both taxa, we hypothesise that the PPGs of ants and beewolves have a common evolutionary origin. Thus, our results suggest that the PPG in ants might not have evolved in response to social requirements but might have already existed in solitary predecessors.     

Is the postpharyngeal gland of a solitary digger wasp homologous to ants? Evidence from chemistry and physiology

The postpharyngeal gland (PPG) was thought to be restricted to ants where it serves a crucial function in the generation of the colony odour. Recently, head glands that closely resemble the PPG of ants were discovered in females of a solitary digger wasp, the European beewolf. The function of this gland necessarily differs from ants: beewolf females apply the secretion of their PPG onto the bodies of paralysed honeybees that serve as larval provisions in order to delay fungus growth. Since ants and digger wasps are not closely related, the occurrence of this gland in these two taxa might either be due to convergent evolution or it is a homologous organ inherited from a common ancestor. Here we test the hypothesis that the PPGs of both taxa are homologous by comparing characteristics of chemical composition and physiology of the PPG of beewolves and ants. Based on reported characteristics of the PPG content of ants, we tested three predictions that were all met. First, the PPG of beewolves contained mainly long-chain hydrocarbons and very few compounds with functional groups. Second, the composition of hydrocarbons in the beewolf PPG was similar to that of the hemolymph. Taking the structure of the gland epithelium and the huge requirements of beewolf females for gland secretion into account this result suggests that the content of the PPG is also sequestered from the hemolymph in beewolves. Third, the chemical composition of the PPG and the cuticle was similar in beewolves since cuticular hydrocarbons derive either from the hemolymph or the PPG. Taking the considerable morphological similarities into account, our results support the hypothesis of a homologous origin of the PPG in beewolves and ants.     

The behaviour of Philanthus crabroniformis (Hymenoptera: Sphecidae)

Observations were made over a two year period on the nesting and hunting behaviour of a population of Philanthus crabroniformis Smith. Data are presented on the time and speed of provisioning, the capture of prey and the species taken, the design and location of nests, the approach to the nest with prey, and aggressive interactions between females. P. gibbosus Fabr., also nested in the same area and took much the same prey. Unlike other sympatric pairs of Philanthus which have been studied, the two species in question were not totally segregated by separate nesting seasons nor by mutually exclusive habitat preferences. One aspect of the behaviour of the two Philanthus which was highly distinctive was the manner in which prey-laden females approached their nests. Selection for divergence in approach patterns may have been exerted by miltogrammine flies, a major parasite of many digger wasps.     

中国榕小蜂触角感受器形态特征及进化适应性分析

【目的】榕树与传粉榕小蜂体系是协同进化、专性传粉的经典系统,每种榕树上一般还生活有种类丰富、数目众多的非传粉榕小蜂。在选择压力下,榕小蜂为在榕果内生存产生了明显的适应性形态。触角感受器是昆虫通讯系统的单元,其形态是反映生态和进化适应性的最佳特征之一。本文旨在对中国部分传粉和非传粉榕小蜂的触角感受器的形态多样性和进化适应性进行系统研究。【方法】对来源于海南和云南15种榕树上24种榕小蜂54个型的触角感受器形态进行了扫描电镜观察,基于现有的分子系统发育树,对传粉榕小蜂触角感受器的形态特征进行了性状演化分析,并对榕小蜂的形态特征进行了进化适应性分析。【结果】榕小蜂触角感受器普遍存在雌雄二型现象。雌蜂触角感受器种类有毛状、锥状、板状、刺状、钟形、腔锥状和栓锥状等,数目丰富,并且进果产卵的传粉雌蜂和果外产卵的非传粉雌蜂之间、进果产卵的传粉雌蜂和进果产卵的非传粉雌蜂之间在形态上存在差异。传粉雄蜂和不具有雄性多型现象的非传粉雄蜂触角感受器极为退化,具有雄性多型的非传粉雄蜂触角感受器形态在种内不具有显著差异。性状演化分析表明进化路径相当复杂,可能存在多次独立进化过程。触角感受器的形态类型与其进化适应性相关。【结论】榕小蜂触角感受器类型多样,形态变化丰富,并为适应榕果内的生存而产生了进化适应性特点。雌蜂和雄蜂在榕果内受到了完全不同的选择压力,行使不同的生态功能,从而产生了不同的适应性形态。不同的适应性形态可能与雌蜂不同的产卵行为、雄蜂不同的交配策略具有一定联系。该文是首次对中国榕小蜂触角感受器形态进行系统研究的报道,有助于更好地理解榕小蜂的形态特征、进化路线、行为策略和生态关系。     

Chemical analyses of wasp-associated streptomyces bacteria reveal a prolific potential for natural products discovery

Identifying new sources for small molecule discovery is necessary to help mitigate the continuous emergence of antibiotic-resistance in pathogenic microbes. Recent studies indicate that one potentially rich source of novel natural products is Actinobacterial symbionts associated with social and solitary Hymenoptera. Here we test this possibility by examining two species of solitary mud dauber wasps, Sceliphron caementarium and Chalybion californicum. We performed enrichment isolations from 33 wasps and obtained more than 200 isolates of Streptomyces Actinobacteria. Chemical analyses of 15 of these isolates identified 11 distinct and structurally diverse secondary metabolites, including a novel polyunsaturated and polyoxygenated macrocyclic lactam, which we name sceliphrolactam. By pairing the 15 Streptomyces strains against a collection of fungi and bacteria, we document their antifungal and antibacterial activity. The prevalence and anti-microbial properties of Actinobacteria associated with these two solitary wasp species suggest the potential role of these Streptomyces as antibiotic-producing symbionts, potentially helping defend their wasp hosts from pathogenic microbes. Finding phylogenetically diverse and chemically prolific Actinobacteria from solitary wasps suggests that insect-associated Actinobacteria can provide a valuable source of novel natural products of pharmaceutical interest.     

The evolution of antennal courtship in diplazontine parasitoid wasps (Hymenoptera,Ichneumonidae, Diplazontinae)

Background

As predicted by theory, traits associated with reproduction often evolve at a comparatively high speed. This is especially the case for courtship behaviour which plays a central role in reproductive isolation. On the other hand, courtship behavioural traits often involve morphological and behavioural adaptations in both sexes; this suggests that their evolution might be under severe constraints, for instance irreversibility of character loss. Here, we use a recently proposed method to retrieve data on a peculiar courtship behavioural trait, i.e. antennal coiling, for 56 species of diplazontine parasitoid wasps. On the basis of a well-resolved phylogeny, we reconstruct the evolutionary history of antennal coiling and associated morphological modifications to study the mode of evolution of this complex character system.

Results

Our study reveals a large variation in shape, location and ultra-structure of male-specific modifications on the antennae. As for antennal coiling, we find either single-coiling, double-coiling or the absence of coiling; each state is present in multiple genera. Using a model comparison approach, we show that the possession of antennal modifications is highly correlated with antennal coiling behaviour. Ancestral state reconstruction shows that both antennal modifications and antennal coiling are highly congruent with the molecular phylogeny, implying low levels of homoplasy and a comparatively low speed of evolution. Antennal coiling is lost on two independent occasions, and never reacquired. A zero rate of regaining antennal coiling is supported by maximum parsimony, maximum likelihood and Bayesian approaches.

Conclusions

Our study provides the first comparative evidence for a tight correlation between male-specific antennal modifications and the use of the antennae during courtship. Antennal coiling in Diplazontinae evolved at a comparatively low rate, and was never reacquired in any of the studied taxa. This suggests that the loss of antennal coiling is irreversible on the timescale examined here, and therefore that evolutionary constraints have greatly influenced the evolution of antennal courtship in this group of parasitoid wasps. Further studies are needed to ascertain whether the loss of antennal coiling is irreversible on larger timescales, and whether evolutionary constraints have influenced courtship behavioural traits in a similar way in other groups.

     

Hemicellulose-degrading bacteria and yeasts from the termite gut

Termites play a major role in the recycling of photosynthetically fixed carbon. With the aid of their symbiotic intestinal flora, they are able to degrade extensively wood constituents such as cellulose and hemicellulose. Nevertheless, the microbial species involved in the degradation of hemicelluloses are poorly defined. The purpose of this paper was to examine the microflora involved in hemicellulose degradation. Different aerobic and facultatively anaerobic bacteria and yeasts were isolated using xylan, arabinogalactan and carboxymethylcellulose as substrates. Gram-positive isolates belonged to the genera Bacillus, Paenibacillus, Streptomyces or the actinobacteria group, while the Gramnegative strains were assigned to the genera Pseudomonas, Acinetobacter, Ochrobactrum , and to genera belonging to the family Enterobacteriaceae. The spectrum and activity of xylan- and arabinogalactan-hydrolysing glycosidases of these new isolates, together with additional bacterial strains originally obtained from enrichments with aromatic compounds were determined.     

Intraspecific specialization of the generalist parasitoid Cotesia sesamiae revealed by polyDNAvirus polymorphism and associated with different Wolbachia infection

As a result of an intense host-parasite evolutionary arms race, parasitic wasps frequently display high levels of specialization on very few host species. For instance, in braconid wasps very few generalist species have been described. However, within this family, Cotesia sesamiae is a generalist species that is widespread in sub-Saharan Africa and develops on several lepidopteran hosts. In this study, we tested the hypothesis that C.?sesamiae may be a cryptic specialist when examined at the intraspecific level. We sequenced exon 2 of CrV1, a gene of the symbiotic polyDNAvirus that is integrated into the wasp genome and is associated with host immune suppression. We found that CrV1 genotype was more closely associated with the host in which the parasitoid developed than any abiotic environmental factor tested. We also tested a correlation between CrV1 genotype and an infection with Wolbachia bacteria, which are known for their ability to induce reproductive isolation. The Wolbachia bacteria infection polymorphism was also found as a major factor explaining the genetic structure of CrV1, and, in addition, the best model explaining CrV1 genetic structure involved an interaction between Wolbachia infection and host species. We suggest that Wolbachia could act as an agent capable of maintaining advantageous alleles for host specialization in different populations of C.?sesamiae. This mechanism could be applicable to other insect models because of the high prevalence of Wolbachia in insects.     

Diverse bacteria related to the bacteroides subgroup of the CFB phylum within the gut symbiotic communities of various termites

Phylogenetically diverse clones of the partial 16S rDNA (ca. 850 bp) of bacteria belonging to the bacteroides subgroup of the cytophaga-flavobacter-bacteroides phylum were collected from the symbiotic microbial communities in the guts of six termite species without cultivation. Combined with the sequences reported previously, a total of thirty phylotypes of the subgroup were identified and classified into five phylogenetic clusters. One that was comprised of the phylotypes from a single termite species was related to the genus Rikenella. Two were clustered each with some cultured strains, genera of which have not been clearly defined yet. The remaining two clusters had no culturable representatives, suggesting the presence of yet-uncultivated genera within the termite guts. From these sequence data, we designed a specific primer for the bacteroides subgroup, which was successful in the terminal-restriction fragment length polymorphism analysis to detect the phylotypes of the subgroup in the termite gut.     

Physiological characterization of the Na(+)/Ca(2+) exchanger (NCX) in hepatopancreatic and antennal gland basolateral membrane vesicles isolated from the freshwater crayfish Procambarus clarkii

The purpose of this study was to physiologically characterize the basolateral Na(+)/Ca(2+) exchanger (NCX) in basolateral membrane vesicles (BLMVs) of hepatopancreas and antennal gland of intermolt crayfish. Conditions were optimized to measure Na(+)-dependent Ca(2+) uptake and retention in the BLMV including use of intravesicular (IV) oxalate and measuring initial uptake rates at 20 s. Na(+)-dependent Ca(2+) uptake rate into BLMV was temperature insensitive. Na(+)-dependent Ca(2+) uptake rate was dependent upon free Ca(2+) with saturable Michaelis-Menten kinetics determined as follows: hepatopancreas, maximal uptake rate (J(max))=2.45 nmol/mg per min, concentration at which carrier operates at half-maximal uptake rate (K(m))=0.69 microM Ca(2+); antennal gland, J(max)=13.2 nmol/mg per min, K(m)=0.59 microM Ca(2+). The two vesicle populations exhibited different sensitivity to putative NCX inhibitors. Benzamil had no effect on Na(+)-dependent Ca(2+) uptake rate in hepatopancreas; in antennal gland it was inhibitory at concentrations up to 30 microM and was stimulatory at higher concentrations. Conversely the inhibitor quinacrine was inhibitory at 10 microM in hepatopancreas and was stimulatory at 1000 microM; meanwhile it was ineffective in antennal gland BLMV. Short circuiting the BLMV had no effect on Na(+)-dependent Ca(2+) uptake rate suggesting that the process may be electroneutral. Compared with another prominent basolateral transporter in hepatopancreas the plasma membrane Ca(2+) ATPase (PMCA), the NCX has 70-fold greater J(max) (at comparable temperature) and a lower affinity. In antennal gland the NCX has 40-fold greater J(max) and a lower affinity. In hepatopancreas and antennal gland BLMV NCX appears to determine the rate of basolateral Ca(2+) efflux in intermolt.