Extended mutualism between termites and gut microbes: nutritional symbionts contribute to nest hygiene

All higher eukaryotes have established symbiotic relationships with diverse microorganisms. One of the most well-characterized symbiotic systems is that of termites and their intestinal microorganisms, which digest cellulose. Recently, diverse types of symbioses between gut microbes and host organisms including humans have received growing attention for various features of their complex interactions beyond nutrition. In termites, researchers are beginning to explore such function of gut symbionts, but only the contribution to internal immunity against entomopathogen is known in a few species. Here, we report that gut symbionts of the dampwood termite Zootermopsis nevadensis protect nests from the spread of the commensal bacterium Serratia marcescens, which has pathogenic potential. Defaunated termites dispersed S. marcescens in the surrounding environment by feeding on the bacteria, which then survived passage through their alimentary tracts, while non-defaunated termites did not. Loss of gut symbionts caused a significant reduction in intestinal acetate, which is an important carbon source for termites. Culture experiments showed that acetate had significant inhibitory effects on S. marcescens at a concentration as low as 12 mM, which indicated that the intestinal acetate of non-defaunated termites (40–130 mM) was capable of suppressing this bacterium. These results suggest that digestive derivatives produced by intestinal symbionts play an essential role in nest hygiene in addition to their nutritional function for termites. Our study provides a better understanding of the multifunctionality of symbiotic relationships in diverse organisms beyond nutrition.     

A complex picture of associations between two host mussels and symbiotic bacteria in the Northeast Atlantic

Among chemosymbiotic metazoans found at deep-sea hydrothermal vents, cold seeps and organic falls, members of the mussel clade Bathymodiolinae (Bivalvia: Mytilidae) have evolved interactions with a higher diversity of bacterial lineages than other bivalve groups. Here, we characterized the bacteria associated with “Bathymodiolus” mauritanicus and Idas-like specimens from three sites in the Northeast Atlantic (two mud volcanoes in the Gulf of Cadiz and one seamount of the Gorringe Bank). Phylogenetic analysis of bacterial 16S rRNA-encoding gene sequences demonstrated that “B”. mauritanicus has a dual symbiosis dominated by two phylotypes of methane-oxidising bacteria and a less abundant phylotype of a sulphur-oxidising bacterium. The latter was the dominant phylotype in a sympatric population of Idas-like mussels at the Darwin mud volcano. These results are the first report of a bacterial phylotype shared between two deep-sea mussels from divergent clades. This sulphur-oxidising bacterium was absent from Idas-like specimens from the other two sites (Gorringe Bank and Meknès mud volcano), in which bacterial clone libraries were dominated by other Gammaproteobacteria related to symbionts previously identified in Idas modiolaeformis from the Eastern Mediterranean. All Idas-like specimens studied herein are closely related and also related to I. modiolaeformis. However, they probably display different associations with bacteria, with the possible absence of both methane- and sulphur-oxidising symbionts at the Gorringe Bank. These results draw a very complex picture of associations between mussels and bacteria in the Northeast Atlantic, which could be highly variable depending on locale characteristics of the habitats.     

Non-specific association between filamentous bacteria and fungus-growing ants

Fungus-growing ants and their fungal cultivar form a highly evolved mutualism that is negatively affected by the specialized parasitic fungus Escovopsis. Filamentous Pseudonocardia bacteria occurring on the cuticle of attine ants have been proposed to form a mutualistic interaction with these ants in which they are vertically transmitted (i.e. from parent to offspring colonies). Given a strictly vertical transmission of Pseudonocardia, the evolutionary theory predicts a reduced genetic variability of symbionts among ant lineages. The aim of this study was to verify whether actinomycetes, which occur on Acromyrmex octospinosus leaf-cutting ants, meet this expectation by comparing their genotypic variability with restriction fragment length polymorphisms. Multiple actinomycete strains could be isolated from both individual ant workers and colonies (one to seven strains per colony). The colony specificity of actinomycete communities was high: Only 15% of all strains were isolated from more than one colony, and just 5% were present in both populations investigated. Partial sequencing of 16S ribosomal deoxyribonucleic acid of two of the isolated strains assigned both of them to the genus Streptomyces. Actinomycetes could also be isolated from workers of the two non-attine ant species Myrmica rugulosa and Lasius flavus. Sixty-two percent of the strains derived from attine ants and 80% of the strains isolated from non-attine ants inhibited the growth of Escovopsis. Our data suggest that the association between attine ants and their actinomycete symbionts is less specific then previously thought. Soil-dwelling actinomycetes may have been dynamically recruited from the environment (horizontal transmission), probably reflecting an adaptation to a diverse community of microbial pathogens.     

Innate humoural immunity is related to eggshell bacterial load of European birds: a comparative analysis

Fitness benefits associated with the development of a costly immune system would include not only self-protection against pathogenic microorganisms but also protection of host offspring if it reduces the probability and the rate of vertical transmission of microorganisms. This possibility predicts a negative relationship between probabilities of vertical transmission of symbionts and level of immune response that we here explore inter-specifically. We estimated eggshell bacterial loads by culturing heterotrophic bacteria, Enterococcus, Staphylococcus and Enterobacteriaceae on the eggshells of 29 species of birds as a proxy of vertical transmission of bacteria from mother to offspring. For this pool of species, we also estimated innate immune response (natural antibody and complement (lysis)) of adults, which constitute the main defence against bacterial infection. Multivariate general linear models revealed the predicted negative association between natural antibodies and density of bacteria on the eggshell of 19 species of birds for which we sampled the eggs in more than one nest. Univariate analyses revealed significant associations for heterotrophic bacteria and for Enterobacteriaceae, a group of bacteria that includes important pathogens of avian embryos. Therefore, these results suggest a possible trans-generational benefit of developing a strong immune system by reducing vertical transmission of pathogens.     

Microbial reduction of uranium (VI) by Bacillus sp. dwc-2: A macroscopic and spectroscopic study

The microbial reduction of U(VI) by Bacillus sp. dwc-2, isolated from soil in Southwest China, was explored using transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and X-ray absorption near edge spectroscopy (XANES). Our studies indicated that approximately 16.0% of U(VI) at an initial concentration of 100 mg/L uranium nitrate could be reduced by Bacillus sp. dwc-2 at pH 8.2 under anaerobic conditions at room temperature. Additionally, natural organic matter (NOM) played an important role in enhancing the bioreduction of U(VI) by Bacillus sp. dwc-2. XPS results demonstrated that the uranium presented mixed valence states (U(VI) and U(IV)) after bioreduction, which was subsequently confirmed by XANES. Furthermore, the TEM and high resolution transmission electron microscopy (HRTEM) analysis suggested that the reduced uranium was bioaccumulated mainly within the cell and as a crystalline structure on the cell wall. These observations implied that the reduction of uranium may have a significant effect on its fate in the soil environment in which these bacterial strains occur.     

Methane Ice Worms: Hesiocaeca methanicola Colonizing Fossil Fuel Reserves

 During a research cruise in July 1997 in the Gulf of Mexico we discovered a gas hydrate approximately 1 m thick and over 2 m in diameter which had recently breached the sea floor at a depth of 540 m. The hydrate surface visible from the submarine was considerably greater than that of any other reported hydrate. Two distinct color bands of hydrate were present in the same mound, and the entire exposed surface of the hydrate was infested (2500 individuals/m²) with 2 to 4 cm-long worms, since described as a new species, Hesiocaeca methanicola, in the polychaete family Hesionidae (Desbruyères and Toulmond 1998). H. methanicola tissue stable isotope values are consistent with a chemoautotrophic food source. No evidence of chemoautotrophic symbionts was detected, but geochemical data support the presence of abundant free living bacteria on the hydrate. The activities of the polychaetes, grazing on the hydrate bacteria and supplying oxygen to their habitats, appears to contribute to the dissolution of hydrates in surface sediments. Received: 16 August 1999 / Accepted in revised form: 19 January 2000     

Response of the bird cherry-oat aphid (Rhopalosiphum padi) to climate change in relation to its pest status, vectoring potential and function in a crop-vector-virus pathosystem

Global climate change threatens world food production via direct effects on plant growth and alterations to pest and pathogen prevalence and distribution. Complex relationships between host plant, pest, pathogen and environment create uncertainty particularly involving vector-borne diseases. We attempt to improve the understanding of the effects of climate change via a detailed review of one crop-vector-pathogen system.The bird cherry-oat aphid, Rhopalosiphum padi, is a global pest of cereals and vector of yellow dwarf viruses that cause significant crop losses in cereals. R. padi exhibits both sexual and parthenogenetic reproduction, alternating between crops and other host plants. In Australia, only parthenogenesis occurs due to the absence of the primary host, thus the aphid continuously cycles from grasses to cereals, allowing for continuous virus acquisition and transmission.We have reviewed the potential impact of future climate projections on R. padi population dynamics, persistence, abundance, dispersal and migration events as well as the interactions between vector, virus, crop and environment, all of which are critical to the behaviour and development of the vector and its ability to transmit the virus. We identify a number of knowledge gaps that currently limit efforts to determine how this pathosystem will function in a future climate.     

Recilia banda Kramer (Hemiptera: Cicadellidae), a vector of Napier stunt phytoplasma in Kenya

Napier grass (Pennisetum purpureum) is the most important fodder crop in smallholder dairy production systems in East Africa, characterized by small zero-grazing units. It is also an important trap crop used in the management of cereal stemborers in maize in the region. However, production of Napier grass in the region is severely constrained by Napier stunt disease. The etiology of the disease is known to be a phytoplasma, 16SrXI strain. However, the putative insect vector was yet unknown. We sampled and identified five leafhopper and three planthopper species associated with Napier grass and used them as candidates in pathogen transmission experiments. Polymerase chain reaction (PCR), based on the highly conserved 16S gene, primed by P1/P6-R16F2n/R16R2 nested primer sets was used to diagnose phytoplasma on test plants and insects, before and after transmission experiments. Healthy plants were exposed for 60 days to insects that had fed on diseased plants and acquired phytoplasma. The plants were then incubated for another 30 days. Nested PCR analyses showed that 58.3% of plants exposed to Recilia banda Kramer (Hemiptera: Cicadellidae) were positive for phytoplasma and developed characteristic stunt disease symptoms while 60% of R. banda insect samples were similarly phytoplasma positive. We compared the nucleotide sequences of the phytoplasma isolated from R. banda, Napier grass on which these insects were fed, and Napier grass infected by R. banda, and found them to be virtually identical. The results confirm that R. banda transmits Napier stunt phytoplasma in western Kenya, and may be the key vector of Napier stunt disease in this region.     

Comparative study on estimation methods for NOx emissions from a roadway

In order to estimate NO_x concentration at road side areas, it is necessary to determine the NO_x emissions from the automobile passing a road. There are two methods for estimation of NO_x emission. One is calculated by the emission factor of automobile, driving mode and traffic volume. Another is a method based on the relation between NO_x and tracer gas concentration of diffusion experiment.First, diffusion experiments by air tracer were conducted. They were carried out at two areas, one of which was an open area and the other was a highly-built-up area. In the experiment, traffic volume, NO_x and tracer gas concentrations were measured simultaneously. However, the result obtained at the former area was used for this analysis.The NO_x emission rate of the automobile was calculated from the data of traffic survey and tracer gas experiments and both results were compared. Three methods for estimation of NO_x emission rate from traffic survey were employed in this study.From these results, it can be confirmed that both results are in agreement and well correlated, so these estimation methods are considered to be appropriate.     

Two selenium tolerant Lysinibacillus sp. strains are capable of reducing selenite to elemental Se efficiently under aerobic conditions

Microbes play important roles in the transport and transformation of selenium (Se) in the environment, thereby influencing plant resistance to Se and Se accumulation in plant. The objectives are to characterize the bacteria with high Se tolerance and reduction capacity and explore the significance of microbial origins on their Se tolerance, reduction rate and efficiency. Two bacterial strains were isolated from a naturally occurred Se-rich soil at tea orchard in southern Anhui Province, China. The reduction kinetics of selenite was investigated and the reducing product was characterized using scanning electron microscopy and transmission electron microscopy-energy dispersive spectroscopy. The bacteria were identified as Lysinibacillus xylanilyticus and Lysinibacillus macrolides, respectively, using morphological, physiological and molecular methods. The results showed that the minimal inhibitory concentrations (MICs) of selenite for L. xylanilyticus and L. macrolides were 120 and 220 mmol/L, respectively, while MICs of selenate for L. xylanilyticus and L. macrolides were 800 and 700 mmol/L, respectively. Both strains aerobically reduced selenite with an initial concentration of 1.0 mmol/L to elemental Se nanoparticles (SeNPs) completely within 36 hr. Biogenic SeNPs were observed both inside and outside the cells suggesting either an intra- or extracellular reduction process. Our study implied that the microbes from Se-rich environments were more tolerant to Se and generally quicker and more efficient than those from Se-free habitats in the reduction of Se oxyanions. The bacterial strains with high Se reduction capacity and the biological synthesized SeNPs would have potential applications in agriculture, food, environment and medicine.     

Determination of estrogenic activity in the river Chienti (Marche Region, Italy) by using in vivo and in vitro bioassays

Environmental estrogen-like compounds (i.e. xenoestrogens) are a variety of pollutants, ranging from synthetic to natural occurring molecules, that are found in surface and waste waters over a wide range of concentrations. In aquatic environment, the overall estrogenic activity is often due to the presence of a mixture of chemicals and their degraded products which can induce synergistic effects. Current strategies for monitoring estrogen-like chemicals are based on the use of a battery of in vivo and in vitro ecotoxicological tests. In this regard, the aim of the present work was to carry out a bio-monitoring study for testing estrogenicity of the Chienti river (Marche Region, Italy) by using both an E-screen and a vitellogenin (Vtg) induction assay in juvenile goldfish. Three sites were used for analysis, localized at the mouth (sampling point 1), in the middle (sampling point 2) and at the origin (sampling point 3) of Chienti river. For most of the water samples (i.e. samples collected at sampling points 2 and 3), clear estrogenic activity was detected in the E-screen assay suggesting different proliferation activities in function of the collecting site. In contrast, the Vtg ELISA demonstrated that water samples collected from each sampling point were estrogenic. Overall, we showed for the first time that the estrogenic activities in water samples from the Chienti river were significant in both in vivo and in vitro; we also observed a different sensitivity between bioassays.     

给水管网中耐氯分枝杆菌的灭活特性及机制研究

近年来,在饮用水管网中检测到耐受消毒剂的微生物,对于饮用水安全造成威胁.课题组由南方某城市自来水管网中分离出1株耐氯的产黏液分枝杆菌(Mycobacteria mucogenicum),并对其灭活特性和耐氯机制进行了研究.使用自由氯、一氯胺和二氧化氯对其进行消毒实验,测定CT值.99.9%灭活产黏液分枝杆菌时,自由氯的CT值为(76.25±47.55)mg·min·L-1,一氯胺为(1 396±382)mg·min·L-1,二氧化氯为(13.5±4.9)mg·min·L-1.采用透射电镜对产黏液分枝杆菌的消毒过程进行观察,发现消毒后细菌结构疏松,细胞器层次变得不清晰,核心肿胀溶解.产黏液分枝杆菌具有较好的疏水性,测定其表面疏水率为37.2%,远高于其他细菌,使得亲水性消毒剂不易进入细菌内部,是其耐受含氯消毒剂的原因之一.     

Bioaccumulation characterization of uranium by a novel Streptomyces sporoverrucosus dwc-3

The biosorption mechanisms of uranium on an aerobic bacterial strain Streptomyces sporoverrucosus dwc-3, isolated from a potential disposal site for (ultra-)low uraniferous radioactive waste in Southwest China, were evaluated by using transmission electron microscopy (TEM), energy dispersive X-ray (EDX) analysis, Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), proton induced X-ray emission (PIXE) and enhanced proton backscattering spectrometry (EPBS). Approximately 60% of total uranium at an initial concentration of 10 mg/L uranium nitrate solution could be absorbed on 100 mg S. sporoverrucosus dwc-3 with an adsorption capacity of more than 3.0 mg/g (wet weight) after 12 hr at room temperature at pH 3.0. The dynamic biosorption process of S. sporoverrucosus dwc-3 for uranyl ions was well described by a pseudo second-order model. S. sporoverrucosus dwc-3 could accumulate uranium on cell walls and within the cell, as revealed by SEM and TEM analysis as well as EDX spectra. XPS and FT-IR analysis further suggested that the absorbed uranium was bound to amino, phosphate and carboxyl groups of the cells. Additionally, PIXE and EPBS results confirmed that ion exchange also contributed to the adsorption process of uranium.     

Effect of pretreatment on Pd/Al2O3 catalyst for catalytic oxidation of o-xylene at low temperature

The effect of pretreatment on Pd/Al₂O₃ catalysts for the catalytic oxidation of o-xylene at low temperature was studied by changing the pretreatment and testing conditions. The fresh and pretreated Pd/Al₂O₃ catalysts were characterized by transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD). The results showed that the pretreatment dramatically changed the Pd/PdO ratio and then significantly affected the Pd/Al₂O₃ activity; while the pretreatment had not much influence on Pd particle size. The Pd/Al₂O₃ pre-reduced at 300℃/400℃, which has fully reduced Pd species, showed the highest activity; while the fresh Pd/Al₂O₃, which has fully oxidized Pd species, presented the worst performance, indicating the Pd chemical state plays an important role in the catalytic activity for the o-xylene oxidation. It is concluded that metallic Pd is the active species on the Pd/Al₂O₃ catalyst for the catalytic oxidation of o-xylene at low temperature.     

Effect of acid solutions on plants studied by the optical beam deflection method

The optical beam deflection method was applied to study the effects of acid solution on both a terrestial and aquatic plants Egeria and Cerastium, which are common aquatic plant and terrestial weed respectively. A probe beam from a He-Ne laser was passed through a vicinity of a leaf of the plants, which were put in culture dishes filled with acid solutions. Deflection signals of the probe beam were monitored and compared for acid solutions with different pH values. The results of Egria showed that the deflection signals changed dramatically when pH values of acid solutions were 2.0 and 3.0, while little at pH of 4.0 and 5.0. For Cerastium when pH were below 3.0, deflection signals changed greatly with time at the begining. After a certain period of time, deflection signals changed little with time. When pH value was above 4.0, deflection signals of Cerastium were still changing with time even after 20 hours. The results suggested that the damage threshold of pH was between 3.0 and 4.0 for both the land and aquatic plants.     

Examining the “evolution of increased competitive ability” hypothesis in response to parasites and pathogens in the invasive paper wasp Polistes dominula

Successful invaders often become established in new ranges by outcompeting native species. The “evolution of increased competitive ability” hypothesis predicts that invasive species are subjected to less predation and parasitization than sympatric native species, and thus can allocate resources from defence and immunity to growth and fecundity, thereby achieving higher fitness. In this study, we examined whether American invasive Polistes dominula paper wasps have reduced immunocompetence. To explore this scenario, we tested their susceptibility towards parasites and pathogens at both the individual (immune defence) and colony levels, i.e. hygienic behaviour (removal of diseased individuals by nestmates). First, we examined the response to the specific coevolved parasite Xenos vesparum (lost after invasion) in terms of individual host susceptibility and hygienic behaviour. Second, we explored the response against general pathogens by quantifying the bacterial clearance in individual wasps after a challenge with Escherichia coli and hygienic behaviour after a challenge with the fungus Beauveria bassiana. Our results show that American invasive P. dominula have a higher response against X. vesparum at the colony level, but at the individual level their susceptibility is not significantly different from conspecifics of the native range. On the other hand, invasive P. dominula display lower response after a challenge with general pathogens at both the individual and colony levels. While supporting the hypothesis of a reduction of immunocompetence towards general pathogens in invasive species, these findings also suggest that the response against coevolved parasites might follow different evolutionary pathways which are not always easily predictable.     

土壤环境中肠道致病菌的多重PCR检测研究初探

建立同时检测大肠杆菌、沙门氏菌、金黄色葡萄球菌、福氏志贺氏菌、铜绿假单胞菌等5种土壤常见肠道致病菌的多重PCR检测技术,为这些肠道致病菌感染的快速诊断提供实验依据.根据这些肠道病原菌的毒素基因、高度保守基因及特异性基因分别合成5对特异性引物,应用PCR扩增技术对目的菌株进行特异性检测.实验结果表明,5对寡核苷酸引物都具有较高的特异性和专一性,多重PCR检测限达到104cfu·g-1.多重PCR应用于土壤样品分析,极大的缩短了检测时间(仅需3～4h)、降低了检测成本,对控制病原菌的传播具有重要意义,可推广应用于环境监测、水源检测、食品卫生监督、商品检验检疫等领域.     

Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR (PMA-qPCR)

The detection of viable bacteria in wastewater treatment plants(WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide(PMA) combined with the quantitative polymerase chain reaction(PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 μmol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.     

A new ceratopsian dinosaur from the Javelina Formation (Maastrichtian) of West Texas and implications for chasmosaurine phylogeny

Bravoceratops polyphemus gen. et sp. nov. is a large chasmosaurine ceratopsid from the lowermost part of the Javelina Formation (early Maastrichtian) of Big Bend National Park, TX, USA. B. polyphemus has a distinctive narrow snout, a long fenestrate frill, and a fan-shaped median parietal bar with a midline epiparietal on its posterior margin, as well as a symmetrical depression on its dorsal surface at the nexus of the parietal rami. This depression is interpreted to be the attachment point for a second midline epiparietal. This parietal morphology is distinct from that exhibited by Anchiceratops or Pentaceratops. The posterior midline epiparietal in B. polyphemus and its bifurcated quadratojugal–squamosal joint are features shared with the most derived chasmosaurines, Torosaurus and Triceratops. The combination of primitive and derived traits exhibited by B. polyphemus, and its stratigraphic position, is compatible with the gradual transition from basal, to intermediate, to derived chasmosaurines observed throughout the western interior of North America, and with phylogenetic analysis, which suggests that Bravoceratops may be closely related to Coahuilaceratops.