Cladophora (Chlorophyta) spp. harbor human bacterial pathogens in nearshore water of Lake Michigan

Cladophora glomerata, a macrophytic green alga, is commonly found in the Great Lakes, and significant accumulations occur along shorelines during the summer months. Recently, Cladophora has been shown to harbor high densities of the fecal indicator bacteria Escherichia coli and enterococci. Cladophora may also harbor human pathogens; however, until now, no studies to address this question have been performed. In the present study, we determined whether attached Cladophora, obtained from the Lake Michigan and Burns Ditch (Little Calumet River, Indiana) sides of a breakwater during the summers of 2004 and 2005, harbored the bacterial pathogens Shiga toxin-producing Escherichia coli (STEC), Salmonella, Shigella, and Campylobacter. The presence of potential pathogens and numbers of organisms were determined by using cultural methods and by using conventional PCR, most-probable-number PCR (MPN-PCR), and quantitative PCR (QPCR) performed with genus- and toxin-specific primers and probes. While Shigella and STEC were detected in 100% and 25%, respectively, of the algal samples obtained near Burns Ditch in 2004, the same pathogens were not detected in samples collected in 2005. MPN-PCR and QPCR allowed enumeration of Salmonella in 40 to 80% of the ditch- and lakeside samples, respectively, and the densities were up to 1.6 x 10(3) cells per g Cladophora. Similarly, these PCR methods allowed enumeration of up to 5.4 x 10(2) Campylobacter cells/g Cladophora in 60 to 100% of lake- and ditchside samples. The Campylobacter densities were significantly higher (P < 0.05) in the lakeside Cladophora samples than in the ditchside Cladophora samples. DNA fingerprint analyses indicated that genotypically identical Salmonella isolates were associated with geographically and temporally distinct Cladophora samples. However, Campylobacter isolates were genetically diverse. Since animal hosts are thought to be the primary habitat for Campylobacter and Salmonella species, our results suggest that Cladophora is a likely secondary habitat for pathogenic bacteria in Lake Michigan and that the association of these bacteria with Cladophora warrants additional studies to assess the potential health impact on beach users.     

The Evolution of Mutualism in Gut Microbiota Via Host Epithelial Selection

The human gut harbours a large and genetically diverse population of symbiotic microbes that both feed and protect the host. Evolutionary theory, however, predicts that such genetic diversity can destabilise mutualistic partnerships. How then can the mutualism of the human microbiota be explained? Here we develop an individual-based model of host-associated microbial communities. We first demonstrate the fundamental problem faced by a host: The presence of a genetically diverse microbiota leads to the dominance of the fastest growing microbes instead of the microbes that are most beneficial to the host. We next investigate the potential for host secretions to influence the microbiota. This reveals that the epithelium–microbiota interface acts as a selectivity amplifier: Modest amounts of moderately selective epithelial secretions cause a complete shift in the strains growing at the epithelial surface. This occurs because of the physical structure of the epithelium–microbiota interface: Epithelial secretions have effects that permeate upwards through the whole microbial community, while lumen compounds preferentially affect cells that are soon to slough off. Finally, our model predicts that while antimicrobial secretion can promote host epithelial selection, epithelial nutrient secretion will often be key to host selection. Our findings are consistent with a growing number of empirical papers that indicate an influence of host factors upon microbiota, including growth-promoting glycoconjugates. We argue that host selection is likely to be a key mechanism in the stabilisation of the mutualism between a host and its microbiota.     

Diverse Microbiota Identified in Whole Intact Nest Chambers of the Red Mason Bee Osmia bicornis (Linnaeus 1758)

Microbial activity is known to have profound impact on bee ecology and physiology, both by beneficial and pathogenic effects. Most information about such associations is available for colony-building organisms, and especially the honey bee. There, active manipulations through worker bees result in a restricted diversity of microbes present within the colony environment. Microbial diversity in solitary bee nests remains unstudied, although their larvae face a very different situation compared with social bees by growing up in isolated compartments. Here, we assessed the microbiota present in nests and pre-adults of Osmia bicornis, the red mason bee, by culture-independent pyrosequencing. We found high bacterial diversity not comparable with honey bee colonies. We identified a variety of bacteria potentially with positive or negative interactions for bee larvae. However, most of the other diverse bacteria present in the nests seem to originate from environmental sources through incorporated nest building material and stored pollen. This diversity of microorganisms may cause severe larval mortality and require specific physiological or symbiotic adaptations against microbial threats. They may however also profit from such a diverse environment through gain of mutualistic partners. We conclude that further studies of microbiota interaction in solitary bees will improve the understanding of fitness components and populations dynamics.     

Cross-phytogroup assessment of foliar epiphytic mycobiomes

The foliar surface forms one of the largest aboveground habitats on Earth and maintains plant-fungus relationships that greatly affect ecosystem functioning. Despite many studies with particular plant species, the foliar epiphytic mycobiome has not been studied across a large number of plant species from different taxa. Using high-throughput sequencing, we assessed epiphytic mycobiomes on leaf surfaces of 592 plant species in a botanical garden. Plants of angiosperms, gymnosperms, and pteridophytes were involved. Plant taxonomy, leaf side, growing environment, and evolutionary relationships were considered. We found that pteridophytes showed the higher fungal species diversity, stronger mutualistic fungal interactions, and a greater percentage of putative pathogens than gymnosperms and angiosperms. Plant taxonomic group, leaf side, and growing environment were significantly associated with the foliar epiphytic mycobiome, but the similarity of the mycobiomes among plants was not directly related to the distance of the host evolutionary tree. Our results provide a general understanding of the foliar fungal mycobiomes from pteridophytes to angiosperms. These findings will facilitate our understanding of foliar fungal epiphytes and their roles in plant communities and ecosystems.     

Occurrence of Escherichia coli and enterococci in Cladophora (Chlorophyta) in nearshore water and beach sand of Lake Michigan

Each summer, the nuisance green alga Cladophora (mostly Cladophora glomerata) amasses along Lake Michigan beaches, creating nearshore anoxia and unsightly, malodorous mats that can attract problem animals and detract from visitor enjoyment. Traditionally, elevated counts of Escherichia coli are presumed to indicate the presence of sewage, mostly derived from nearby point sources. The relationship between fecal indicator bacteria and Cladophora remains essentially unstudied. This investigation describes the local and regional density of Escherichia coli and enterococci in Cladophora mats along beaches in the four states (Wisconsin, Illinois, Indiana, and Michigan) bordering Lake Michigan. Samples of Cladophora strands collected from 10 beaches (n = 41) were assayed for concentrations of E. coli and enterococci during the summer of 2002. Both E. coli and enterococci were ubiquitous (up to 97% occurrence), with overall log mean densities (+/- standard errors) of 5.3 (+/- 4.8) and 4.8 (+/- 4.5) per g (dry weight). E. coli and enterococci were strongly correlated in southern Lake Michigan beaches (P < 0.001, R(2) = 0.73, n = 17) but not in northern beaches (P = 0.892, n = 16). Both E. coli and enterococci survived for over 6 months in sun-dried Cladophora mats stored at 4 degrees C; the residual bacteria in the dried alga readily grew upon rehydration. These findings suggest that Cladophora amassing along the beaches of Lake Michigan may be an important environmental source of indicator bacteria and call into question the reliability of E. coli and enterococci as indicators of water quality for freshwater recreational beaches.     

Increasing antagonistic interactions cause bacterial communities to collapse at high diversity

Biodiversity is a major determinant of ecosystem functioning. Species-rich communities often use resources more efficiently thereby improving community performance. However, high competition within diverse communities may also reduce community functioning. We manipulated the genotypic diversity of Pseudomonas fluorescens communities, a plant mutualistic species inhibiting pathogens. We measured antagonistic interactions in vitro, and related these interactions to bacterial community productivity (root colonisation) and ecosystem service (host plant protection). Antagonistic interactions increased disproportionally with species richness. Mutual poisoning between competitors lead to a 'negative complementarity effect', causing a decrease in bacterial density by up to 98% in diverse communities and a complete loss of plant protection. The results emphasize that antagonistic interactions may determine community functioning and cause negative biodiversity-ecosystem functioning relationships. Interference competition may thus be an additional key for predicting the dynamics and performance of natural assemblages and needs to be implemented in future biodiversity models.     

Diversity and distribution of epibiotic bacteria on Pseudo-nitzschia multiseries (Bacillariophyceae) in culture, and comparison with those on diatoms in native seawater

Previous studies on the interaction between bacteria and harmful algal bloom species have mostly considered the bacteria in the bulk solution. Here, we document the abundance and mode of attachment of bacteria growing on the cell surface of the domoic acid-producing diatom Pseudo-nitzschia multiseries (Hasle) Hasle in culture, compared with diatoms in field samples. The epiphytic bacteria were examined by scanning electron microscopy to visualize their morphology and mode of attachment. Two P. multiseries cultures were studied: clone CLN-1 and sub-clone CLN-1-NRC; the latter had been maintained in another laboratory for 2 years. Each of these P. multiseries cultures exhibited a clearly different assemblage of epibiotic bacteria, even though both originated from the same parent culture. The bacterial diversity was greater in clone CLN-1 (nine distinct morphotypes seen) than in sub-clone CLN-1-NRC (six morphotypes). The former clone also produced more domoic acid than the latter. There was a succession of bacterial morphotypes as well as an increase in the number of epiphytic bacteria per diatom cell during the progression from exponential to stationary phase. The most diverse and common morphotypes were rod-shaped cells (e.g. a Caulobacter-like bacterium attached by a discoid holdfast). Epibionts showed a preference for attachment at specific regions of the host diatom frustule, e.g. the raphe or cingulum, locations where organic matter may be extruding from the diatom cell. Most diatom cells carried only one to five bacteria, and up to ca. 60% of the intact diatom cells (although intact cells themselves were infrequent) were still free of epibiotic bacteria at the end of the 31-day batch culture experiment. Sequencing of the SSU rRNA gene showed that five of the eight bacterial strains isolated from the P. multiseries cultures were members of the Alphaproteobacteria, three of the Gammaproteobacteria and one of the Bacteroidetes. A morphologically diverse assemblage of epibiotic bacteria was also found on both centric and pennate planktonic diatoms in natural coastal waters. Of the eight morphotypes recorded, all but two were also found in the cultures. Relatively fewer wild diatom cells carried bacteria compared to cells in culture. We hypothesize that the diversity and abundance of epiphytic bacteria may explain some of the variability seen in the production of DA by different P. multiseries clones, and should be considered as another important and controllable variable that influences diatom cell physiology.     

The Molecular Basis of Bacterial–Insect Symbiosis

Insects provide experimentally tractable and cost-effective model systems to investigate the molecular basis of animal–bacterial interactions. Recent research is revealing the central role of the insect innate immune system, especially anti-microbial peptides and reactive oxygen species, in regulating the abundance and composition of the microbiota in various insects, including Drosophila and the mosquitoes Aedes and Anopheles. Interactions between the immune system and microbiota are, however, bidirectional with evidence that members of the resident microbiota can promote immune function, conferring resistance to pathogens and parasites by both activation of immune effectors and production of toxins. Antagonistic and mutualistic interactions among bacteria have also been implicated as determinants of the microbiota composition, including exclusion of pathogens, but the molecular mechanisms are largely unknown. Some bacteria are crucial for insect nutrition, through provisioning of specific nutrients (e.g., B vitamins, essential amino acids) and modulation of the insect nutritional sensing and signaling pathways (e.g., insulin signaling) that regulate nutrient allocation, especially to lipid and other energy reserves. A key challenge for future research is to identify the molecular interaction between specific bacterial effectors and animal receptors, as well as to determine how these interactions translate into microbiota-dependent signaling, metabolism, and immune function in the host.     

绿球藻目的几个中国新记录属种

6个生境比较特殊而有趣的绿球藻目物种首次在我国发现,它们分别是生于土壤的香喇网绿藻(Dictyochloris fragrans Vischer ex Start),寄生于眼子菜叶片的浮萍绿点藻(Chlorchytrium lemmae Cohn.),气生种类—生长在樟树皮上的分层胶囊藻(Gloeocystis polydermatica(Kutz.) Hindak),与真菌共生形成地衣的土著共球藻(Trebouxia aboricola de Puymaly)和团集共球藻(Trebouscia glomerata(Waren) Ahmadjian),还有附生于一种刚毛藻(Cladophora sp.)细胞表面的伊乐外生藻(Ectogeron elodeae Dangeard)。除胶囊藻属(Gloeocystis)外,其它属均为中国新记录属。     

Wide bacterial diversity associated with tubes of the vent worm Riftia pachyptila

We carried out a 16S rDNA-based molecular survey of the prokaryotic diversity associated with the chitin tubes of the giant vent tubeworm Riftia pachyptila (collected at the East Pacific Rise, 9 degrees N and 13 degrees N). Scanning electron microscopy showed dense microbial populations, particularly on the external surface of the middle and upper tube regions, which included very diverse prokaryotic morphotypes. We used archaeal- and bacterial-specific primers for polymerase chain reaction (PCR) amplification, but only bacterial amplicons were obtained. We analysed a total of 87 clones. Most belonged to the epsilon-Proteobacteria, but also to the delta-, alpha- and gamma-Proteobacteria. A broad diversity of phylotypes belonging to other bacterial divisions was detected, including Verrucomicrobia, the Cytophaga-Flavobacterium-Bacteroides group and the candidate division OP8. We also retrieved a sequence, R76-B150, of uncertain phylogenetic affiliation, which could represent a novel candidate division. The sequence of the R. pachyptilagamma-proteobacterial endosymbiont was not detected. The bacterial diversity found suggests that complex metabolic interactions, particularly based on sulphur chemistry, may be occurring in different microniches of the R. pachyptila tubes.     

Effects of suspended sediment and desiccation on the benthic tailwater community in the Colorado River, USA

We demonstrated that differences in habitatrequirements by C. glomerata and Oscillatoria havea profound bottom-up influence on the foodweb in thetailwaters below Glen Canyon Dam in the Colorado Riverthrough Grand Canyon National Park, USA. We examinedthe effects of suspended sediment and desiccation onthe colonization sequence of Cladophora glomerata andOscillatoria spp. and the consequent effects onmacroinvertebrates in each algal community in a seriesof reciprocal transplants in the regulated ColoradoRiver, AZ. Our experiments showed that C. glomeratagrows best in continuously submerged, clear-water,stable habitats, whereas Oscillatoria forms densemat-like matrices of trichomes and sand in varialzones and habitats with high suspended sedimentstypical of many southwestern USA streams. Varial zoneconditions have a stronger influence on communitystructure than habitats with high suspended sediments. Recruitment by chironomid larvae was less dependent onC. glomerata and less affected by suspended sedimentand periodic desiccation than Gammarus lacustris. Weestimated the energy from macroinvertebrate biomassassociated with tufts of C. glomerata to be an orderof magnitude higher than that in Oscillatoriamatrices. Therefore, loss of C. glomerata andreplacement of habitat more suitable for Oscillatoriaas a result of regulated flows indirectly reducespotential energy flow in the Colorado Riverfoodweb.     

The Enteric Two‐Step: nutritional strategies of bacterial pathogens within the gut

The gut microbiota is a dense and diverse microbial community governed by dynamic microbe–microbe and microbe–host interactions, the status of which influences whether enteric pathogens can cause disease. Here we review recent insights into the key roles that nutrients play in bacterial pathogen exploitation of the gut microbial ecosystem. We synthesize recent findings to support a five‐stage model describing the transition between a healthy microbiota and one dominated by a pathogen and disease. Within this five‐stage model, two stages are critical to the pathogen: (i) an initial expansion phase that must occur in the absence of pathogen‐induced inflammation, followed by (ii) pathogen‐promoting physiological changes such as inflammation and diarrhoea. We discuss how this emerging paradigm of pathogen life within the lumen of the gut is giving rise to novel therapeutic strategies.     

Facilitation of periphyton production by tadpole grazing: functional differences between species

1. This study examined how interactions between resources that vary in edibility, and herbivores that vary in ability to acquire resources, control primary productivity. In a northern California river, grazing on Cladophora glomerata , a relatively inedible filamentous green alga, and its more nutritious epiphytic diatoms, was manipulated by exposing cobbles to tadpoles ( Rana boylii or Hyla regilla ) or excluding tadpoles.
2. Rana indirectly facilitated Cladophora by removing diatoms, whereas Hyla did not significantly change biomass relative to controls. Algal ash-free dry mass on cobbles in Rana treatments was 65 and 72% greater than on controls in two years of investigation (1991 and 1993). Rana decreased epiphytic diatom biovolume by 56% and detritus by 87%.
3. Because nitrogen excretion rates of Hyla and Rana were similar, the differences in effect between the two species were probably due to their roles as consumers rather than as recyclers.
4. The net effect of Rana on periphyton was a 10% increase in areal specific primary productivity (mg O₂ h^–1 m^–2); Hyla caused an 18% decrease. Rana decreased biomass-specific productivity (mg O₂ h^–1 g^–1) 44%; Hyla had no effect.
5. In tadpole exclosures, grazers such as baetid mayfly larvae (mostly Centroptilum sp.) were 4.7 (1991) and 1.8 (1993) times more abundant, and midge larvae (Chironomidae) were 2.5 (1991) and 2 (1993) times more abundant than in Rana enclosures. Invertebrate assemblages in Hyla enclosures, however, were similar to exclosures. Few predatory insects and fish colonized Rana enclosures. Path analyses indicated that Rana affected macroinvertebrates via both interference and exploitation of epiphytic diatoms.     

Facilitation of periphyton production by tadpole grazing: functional differences between species

1. This study examined how interactions between resources that vary in edibility, and herbivores that vary in ability to acquire resources, control primary productivity. In a northern California river, grazing on Cladophora glomerata , a relatively inedible filamentous green alga, and its more nutritious epiphytic diatoms, was manipulated by exposing cobbles to tadpoles ( Rana boylii or Hyla regilla ) or excluding tadpoles.
2. Rana indirectly facilitated Cladophora by removing diatoms, whereas Hyla did not significantly change biomass relative to controls. Algal ash-free dry mass on cobbles in Rana treatments was 65 and 72% greater than on controls in two years of investigation (1991 and 1993). Rana decreased epiphytic diatom biovolume by 56% and detritus by 87%.
3. Because nitrogen excretion rates of Hyla and Rana were similar, the differences in effect between the two species were probably due to their roles as consumers rather than as recyclers.
4. The net effect of Rana on periphyton was a 10% increase in areal specific primary productivity (mg O₂ h^–1 m^–2); Hyla caused an 18% decrease. Rana decreased biomass-specific productivity (mg O₂ h^–1 g^–1) 44%; Hyla had no effect.
5. In tadpole exclosures, grazers such as baetid mayfly larvae (mostly Centroptilum sp.) were 4.7 (1991) and 1.8 (1993) times more abundant, and midge larvae (Chironomidae) were 2.5 (1991) and 2 (1993) times more abundant than in Rana enclosures. Invertebrate assemblages in Hyla enclosures, however, were similar to exclosures. Few predatory insects and fish colonized Rana enclosures. Path analyses indicated that Rana affected macroinvertebrates via both interference and exploitation of epiphytic diatoms.     

Plasticity and acclimation to light reflected in temporal and spatial changes of small-scale macroalgal distribution in a stream

The small-scale distribution pattern of macroalgae in the river Ilm, in Germany was monitored. These patterns were then related to abiotic factors and tested to discover whether the distribution of the common macroalgae, Cladophora glomerata (L.) Kütz. and Vaucheria sp., was linked to differences in their photosynthetic plasticity. Cladophora glomerata revealed higher maximum photosynthetic electron transport rates after acclimation to high light (HL) compared with low light (LL) acclimated samples. By contrast, Vaucheria sp. did not acclimate to different growth light conditions. The photosynthetic performance of both algae also varied according to diurnal conditions. High light caused a reversible decrease of the dark-adapted quantum yield (F(v)/F(m)) in C. glomerata and a concomitant reversible decrease of the light-adapted quantum yield (DeltaF/F'(m)). In Vaucheria sp., F(v)/F(m) remained mostly unchanged over the day, whereas DeltaF/F'(m) decreased during the morning at low light. Photosynthetic pigments confirmed acclimational differences between the species. HL C. glomerata showed increased chlorophyll a:chlorophyll b ratios, and higher amounts of xanthophyll-cycle pigments compared with LL samples, whereas Vaucheria sp. did not reveal differences between the light treatments. While preferences for substrate size, water velocity, and depth are similar for C. glomerata and Vaucheria sp., the physiological responses to light conditions are different. It is concluded that light conditions significantly affect the small-scale spatial distribution of macroalgae and that fitness is enhanced in species with a higher plasticity in photosynthetic acclimation in unstable environments.     

ECOLOGY OF CLADOPHORA GLOMERATA (L.) KÜTZ IN SOUTHERN ONTARIO1

Cladophora glomerata (L.) Kütz. was the dominant attached alga in streams and along lake shores in southern Ontario. Maximum production occurred in summer and resulted from 2 short periods of intensive vegetative growth (June and September). Optimum habitat conditions consist of a firm substrate in shallow alkaline water. These conditions are provided by harbor facilities where man-made structures supplement naturally occurring rock outcrops or cobble beaches and where nutrients are available from tributaries draining agricultural and urban land areas. Periodicity appears to be related to temperature; the interval between the 2 periods of intensive growth during warm weather decreases at successively more northern locations. Frequent observation of Cladophora growing at a single stream station revealed that variations in thallus morphology and relative cell dimensions of C. glomerata are greater than has generally been recognized. These variations can be attributed to changes in environmental conditions and events in the annual life cycle of the alga.     

Shifting the balance: antibiotic effects on host-microbiota mutualism

Antibiotics have been used effectively as a means to treat bacterial infections in humans and animals for over half a century. However, through their use, lasting alterations are being made to a mutualistic relationship that has taken millennia to evolve: the relationship between the host and its microbiota. Host-microbiota interactions are dynamic; therefore, changes in the microbiota as a consequence of antibiotic treatment can result in the dysregulation of host immune homeostasis and an increased susceptibility to disease. A better understanding of both the changes in the microbiota as a result of antibiotic treatment and the consequential changes in host immune homeostasis is imperative, so that these effects can be mitigated.     

Interactions between functionally diverse fungal mutualists inconsistently affect plant performance and competition

Plants form mutualistic relationship with a variety of belowground fungal species. Such a mutualistic relationship can enhance plant growth and resistance to pathogens. Yet, we know little about how interactions between functionally diverse groups of fungal mutualists affect plant performance and competition. We experimentally determined the effects of interaction between two functional groups of belowground fungi that form mutualistic relationship with plants, arbuscular mycorrhizal (AM) fungi and Trichoderma, on interspecific competition between pairs of closely related plant species from four different genera. We hypothesized that the combination of two functionally diverse belowground fungal species would allow plants and fungi to partition their symbiotic relationships and relax plant–plant competition. Our results show that: 1) the AM fungal species consistently outcompeted the Trichoderma species independent of plant combinations; 2) the fungal species generally had limited effects on competitive interactions between plants; 3) however, the combination of fungal species relaxed interspecific competition in one of the four instances of plant–plant competition, despite the general competitive superiority of AM fungi over Trichoderma. We highlight that the competitive outcome between functionally diverse fungal species may show high consistency across a broad range of host plants and their combinations. However, despite this consistent competitive hierarchy, the consequences of their interaction for plant performance and competition can strongly vary among plant communities.     

Dissolved organic matter utilization and oxygen uptake in algal-bacterial microcosms.

Under closed laboratory conditions, at non-limiting nutrient levels, the biomass of Anabaena variabilis, Anacystis nidulans, Chlorella pyrenoidosa, and Selanastrum capricornutum increased with increasing levels of dissolved organic matter (DOM) as a result of bacterially produced carbon dioxide (CO2) and (or) cofactors. Oxygen (O2) produced as a result of algal photosynthesis was sufficient to supply the najority of O2 required by the bacterial community. The percentage of DOM utilized by bacteria which was subsequently incorporated into algal biomass varied with individual species indicating that the association between individual algal species and the bacterial microbiota varied. Under natural conditions bacteria could provide CO2 and (or) cofactors for algal photosynthesis which in turn supplies O2 for bacterial respiration. This mutualistic association in aquatic environments could result in an increase in planktonic and epiphytic algal biomass if other nutrients are available.