16S RNA Gene Sequencing Research Articles

Impact of temperature stress on Pyropia yezoensis and its inhabitant microbiota to promote aquaculture

The diverse inhabitant microbiota of Pyropia yezoensis can establish both beneficial and harmful interactions that effect community structure. It is complex to understand the seaweed microbial diversity and their ecological and functional regulations which are affected by numerous environmental factors. In this study, temperature stress was focused to understand the metagenomics of P. yezoensis microbiota, its functional annotation and the metabolites regulation of P. yezoensis. The samples of P. yezoensis from yellow sea of China were divided into two groups i.e low temperature 5°C (LT) and High temperature 25°C (HT). The genes and functional level analysis were followed by using metagenomic sequencing analysis and sequencing of 16sRNA gene. The Pyropia yezoensis samples were also analyzed through LCMS for metabolites regulation due to effects of microbiota and temperature stress. According to taxonomic classification analysis, Proteobacteria, Bacteroidetes, and Firmicutes were predominant in both samples with over-representation of these species in high temperature stress samples. A diverse relationship between the bacterial communities residing in the LT and HT samples was explored through discriminant analysis. The functional prediction of bacterial communities from KEGG analysis revealed a notable increase in HT predicted genes associated to amino acid metabolism, carbohydrate metabolism, energy metabolism, co-factors and vitamins metabolism, membrane transport, and nucleotide metabolism. The provoked chemical signaling and transportation response from the bacterial community, leading to activation of metabolic genes to utilize the available substrates of seaweed. The LCMS illustrated that the primary metabolites of Pyropia yezoensis were down regulated due to effect of high temperature which indicates a negative effect on the growth and health of Pyropia, while the secondary metabolites were up-regulated showing the defensive mechanism against unbearable stress. These up and down regulated metabolites also activated their respective pathways, suggesting the adoptive mechanism of P. yezoensis in response of high temperature.

Effects of sample collection and storage methods on fecal bacterial diversity in California ground squirrels (Otospermophilus beecheyi)

Abstract Measures of fecal bacterial abundance and taxonomic composition are commonly used as proxies for gut microbial diversity in studies of free-living mammals. Because methods of sample collection and storage may affect measures of bacterial diversity, we evaluated the effects of several procedures on fecal bacterial diversity in a free-living population of California ground squirrels (Otospermophilus beecheyi). Replicate fecal samples from 12 adult female squirrels were collected either from the soil beneath traps in which individuals had been captured or from tubs placed under squirrels during handling. Samples were then either frozen immediately in liquid nitrogen or stored on ice for several hours before being transferred to a −80°C freezer. Sequencing of the bacterial 16s RNA gene revealed no differences among methods with regard to sequence read depth (number of sequences recovered per animal) or alpha (within-individual) diversity of bacterial taxa. While our collection-storage procedures had a significant effect on one of the metrics of beta (among-individual) bacterial diversity examined, this effect was small compared to that of individual identity. Date of sample collection affected alpha and beta diversity; samples collected only 1 week apart differed significantly in bacterial diversity. Overall, these findings suggest that the collection and storage methods employed yield generally comparable information and are capable of capturing potentially important patterns of fecal bacterial composition and diversity in free-living mammals.

Exploring the Correlation between Changes in Gut Microbial Community Diversity and Depression in Human Populations.

Depression, also known as depressive disorder, is a group of psychosomatic affective disorders characterized by persistent and significantly depressed mood, delayed thinking, and cognitive impairment. The aim of this study was to explore the correlation between changes in gut microbial community diversity and depression to provide data on new strategies for the prevention and treatment of depression. In this study, we separated participants into a group of depressed patients and a healthy comparison group. We analyzed the gut microbial community structure of depressed patients and healthy comparisons using second-generation sequencing of the bacterial 16S RNA gene. There were significant differences in the gut microflora structure between patients with depression and healthy individuals. The gut flora alpha diversity index was significantly reduced in patients with depression compared to that in the healthy population. At the species level, the relative abundance of Coprococcus catus and Bacteroides barnesiae was significantly lower in the depressed group than that in the control group. The development of depression may be associated with a decrease in beneficial gut bacteria.

Discovery of Rickettsia and Rickettsiella Intracellular Bacteria in Emerald Ash Borer Agrilus planipennis by Metagenomic Study of Larval Gut Microbiome in European Russia

Emerald ash borer Agrilus planipennis (Coleoptera: Buprestidae) is a quarantine pest posing a threat to ash trees all over Europe. This wood-boring beetle native to Asia is quickly spreading in North America and European Russia, and approaching the European Union and the Middle East. It is important to study microorganisms associated with this pest, because the knowledge of its “natural enemies” and “natural allies” could be potentially used for the control of the pest. All previously published information about the A. planipennis microbiome was obtained in North America and China. We present the first study on procaryotes associated with A. planipennis in Europe. Alive larvae were sampled from under the bark of Fraxinus pennsylvanica in the Moscow Oblast and the gut microbiome was studied using metagenomic methods. Next-generation Illumina-based amplicon sequencing of the v3-v4 region 16S-RNA gene was performed. In total, 439 operational taxonomic units from 39 families and five phyla were detected. The dominant families in our samples were Pseudomonadaceae, Erwiniaceae and Enterobacteriaceae, in accordance with the published information on the larval gut microbiome in North America and China. We detected intracellular bacteria in A. planipennis for the first time, namely Rickettsia (Rickettsiaceae) and Rickettsiella (Diplorickettsiaceae). Representatives of the genus Rickettsia are known to be in mutualistic symbiosis with some phytophagous insects, while Rickettsiella bacteria are pathogenic to many arthropods. The finding of Rickettsia and Rickettsiella opens perspectives for future research on the interactions between these bacteria and A. planipennis and the possible use of these interactions for the control of the pest.

The Negative Effects of Feces-Associated Microorganisms on the Fitness of the Stored Product Mite Tyrophagus putrescentiae.

Feces have been suggested as a major source of microorganisms for recolonization of the gut of stored product mites via coprophagy. The mites can host microorganisms that decrease their fitness, but their transmission is not known. To address the role of fecal microbiota on mite fitness, we performed an experimental study in which the surfaces of mite (Tyrophagus putrescentiae) eggs were sterilized. Mites eggs (15 per experimental box) were then hatched and grown on feedstock with and without feces. These experiments were conducted with four distinct T. putrescentiae populations (5L, 5K, 5N, and 5P), and mite population density after 21 day of cultivation was used to assess mite fitness and the impact of fecal microbiota on fitness. Population density was not affected by the presence of feces in two of the cultures (5L and 5K), while significant effects of feces were observed in the other cultures (5N and 5P). Mite culture microbial communities were analyzed using cultivation-independent next-generation amplicon sequencing of microbial 16S and 18S ribosomal RNA (rRNA) genes in the fitness influenced populations (5N and 5P). Several microbial taxa were associated with fecal treatments and reduced mite fitness, including Staphylococcus and Bartonella-like bacteria, and the fungal genera Yamadazyma, Candida, and Aspergillus. Although coprophagy is the transmission route mites used to obtain beneficial gut bacteria such as Bartonella-like organisms, the results of this study demonstrate that fecal-associated microorganisms can have negative effects on some populations of T. putrescentiae fitness, and this may counteract the positive effects of gut symbiont acquisition.

Comparative Study Between the Sequencing of 16sRNA gene and Traditional Identification Methods of Some Bacterial Pathogens Associated with A Systemic Bacterial Infections from Diseased Poultry Flocks

Poultry health problems of chickens are usually accompanied by bacterial systemic infections causing severe losses in flocks, sick birds were systemically could be infected by several bacteria regardless of the clinical signs. 16s RNA sequence is a modern approach for the diagnosis of bacterial infection, especially of multiple causative agents. Ten field cases suffering from systemic infections have been examined for bacterial causes, using conventional isolation and 16S rRNA sequencing, The ten sequenced samples were compatible with the results of isolation; six samples were similar to E.coli species with identity% = 97-99%, while it had similarity% < 96% with Salmonella enterica. While the other samples showed variable identity ranging from 94-99% with Salmonella enterica, E. coli, Enterobacter spp and Klebsiella spp In conclusion, although conventional isolation is the golden standard method for bacterial diagnosis, however, the 16s RNA sequencing has elaborated excellence in diagnosis and identification of bacterial infection, concerning the time consuming, detection of multi-caused infections and sensitivity.

English

The sheath rot rice disease is known to be associated to Pseudomonas fuscovaginae and a recent study on the symptomatic of sheath rot revealed that this bacterial pathogen is abundant in highland rice ecology cultivation in Burundi. From asymptomatic samples of sheath rice, a collection of bacterial isolates and culturable microbiome have been carried out. A comparison between the culturable microbiome and total microbiome was made. Phenotypic assays in vitro on the bacterial isolates were performed after identification at the genus level of each bacterial isolate by sequencing of 16S RNA gene. A bacterial isolate belonging to Alcaligenes genus has antibacterial activity against the rice sheath rot pathogen P. fuscovaginae in vitro conditions.   Key words: Pseudomonas fuscovaginae, sheath rot, microbiome, Alcaligenes species.

Complex Internal Microstructure of Feather Follicles on Chicken Skin Promotes the Bacterial Cross-Contamination of Carcasses During the Slaughtering Process.

Chicken skin is considered the most susceptible to bacterial contamination during slaughter. It is rich in bushy feather follicles with complex internal structures that can absorb bacteria via cross-contamination during slaughter. Until now, the microstructural changes and local bacterial composition of feather follicles during slaughter have not been thoroughly investigated. This study used hematoxylin-eosin (HE) staining of the tissue paraffin section to investigate the structure of the feather follicles on chicken skin. In addition, the biopsy sampling method was employed for the high-throughput sequencing of 16S RNA genes to study the composition and source of bacterial contamination during slaughter. The results show that the feather follicles on chicken skin form a closed cavity structure during the slaughtering process. The volume of the irregular follicle cavity was about Ø: 200 μm × D: 1040 μm, which provides a place for the bacteria to absorb and resist the cleaning and disinfection during the slaughtering process. The composition of bacteria in the feather follicle was mainly Acinetobacter (37%), Psychrobacter (8%), Macrococcus (5%), and Comamonas (2%). The heat map obtained via the species abundance analysis of the feather follicle samples as well as the slaughter environment samples suggests that the gastrointestinal feces contaminated the feather follicles on the chicken skin mainly during the evisceration, defeathering, and chilling processes, and the last-stage chilling water also caused severe cross-contamination to the feather follicles during the chilling process.

IMPACT OF ENDOPHYTIC LIPOPEPTIDES USE AGAINST SOME PLANT PATHOGENIC MICROORGANISMS

Twelve dissimilar plant associated bacterial colonies were isolated from five different tomato seedlings, the Non Ribosomal Lipopeptides genes were detected by degenerated primers technique in the twelve tomato endophytic isolates, only the long rode bacilli endophytic strains were found to harbour the lipopeptides synthetases genes, these three strains were given laboratory names of (BMG100, BMG101 and BMG102) for further pursuit. Partial sequencing of 16s RNA gene and protein profile analysis for the three bacilli isolates showed that all isolates belonged to Bacillus subtilis. The lipopeptides production by HPLC analysis revealed that all the endophytic isolates from tomato stems had the ability to produce lipopeptides except the third isolate named B. subtilis BMG102 showed no production. different levels of volumetric oxygen transfer coefficient (KLa) were used as cursors for lipopeptides production. The maximum concentration of surfactin type was about 1431, 529 and 437 mg.L-1, the plipastitin (fengycin type ) was 360 198 mg.L-1 and zero while the Mycosubtilin (iturin type) was zero, 228 and 414 mg.L-1 for the strains B. subtilis ATCC21332, BMG100 and BMG101, respectively. Surfactin production is visibly optimal at high oxygen transfer (kLa) and showed more than 0.07 S-1, while the optimum production of the fengycin and iturin could be observed at mild values of O2 transfer (kLa) which equal 0.01 and 0.015 S-1 respectively. The antibacterial - antifungal effects of the three B.subtilis extracts collected during lipopeptides productivity regimes showed inhibition activity against Pseudomonas syringae, Ralstonia solanacearum and Erwinia amylovora, as will as the antifungal activity against Aspergillus niger, Phanerochaete chrysosporium and Alternaria alternata. These results indicate potential use of B.subtilis strains as biocontrol agents, especially when considering commercial implementation. Studies regarding the lipopeptides applications, should also be considered as this will influence the efficacy of the lipopeptides against the target organisms.

Temperature potentially induced distinctive flavor of mud crab Scylla paramamosain mediated by gut microbiota

Many factors affect the flavor of crabs. However, impact of temperature on flavor has not been reported. Here, we examined Scylla paramamosain collected within the main four producing areas in China from north sampling point (NP) and south sampling point (SP), respectively. The contents of flavouring-related substances in hepatopancreas, muscles and gonads were determined by high-performance liquid chromatography (HPLC). Meanwhile, high-throughput sequencing of 16S RNA gene was used to reveal the diversity distribution of gut microbiota at each sample collection point. Comparisons among flavor substances of edible parts, the implied higher temperature in SP may be beneficial to the accumulation of flavor substances in gonads, while lower temperature in NP may be beneficial to the accumulation of flavor substances in muscles and hepatopancreas. The gut microbiota of crabs, was analyzed via 16S rRNA gene sequencing. The results of gut microbiota showed that there were significant differences in the distribution of gut microbiota in NP and SP. The microbiota composition of SP has a high distribution richness and no absolute dominant bacteria, while NP has absolute dominant bacteria and its microbiota richness was lower than SP. The results of redundancy analysis (RDA) showed that there was a significant correlation between temperature and the relative abundance of gut microbiota, and a significant correlation between gut microbiota and the content of flavor substances. This study indicates that temperature may be one of the main factors for the differences of flavor substances between SP and NP, which was most probably mediated by gut microbiota. Further exploration is needed with laboratory experiments in which the environment is more precisely controlled if these views are to be determined.

Detection and characterization of multidrug-resistant Salmonella enterica serovar Infantis as an emerging threat in poultry farms of Faisalabad, Pakistan.

The aim of this study was the molecular identification of Salmonella enterica serovar Infantis (S. Infantis) isolated from poultry samples and their antimicrobial resistance and virulence profiling. A total of 149 isolates, belonging to genus Salmonella, originally isolated from 340 suspected poultry post mortem specimens reported by us earlier were preliminary identified as Salmonella by biochemical methods and confirmed by polymerase chain reaction targeting genus-specific gene invA. Targeting serovar-specific gene fragment (fljB) resulted in confirmation of 54 isolates as S. Infantis which were further confirmed by sequencing of 16S RNA and fljB genes. Swimming and swarming motilities were detected in 98·1 and 11·1% isolates respectively. Phenotypic disc diffusion assay against 23 antimicrobial agents showed the highest resistance against pefloxacin (PEF) (94·4%), chloramphenicol (83·3%) and imipenem (77·7%) while 5·3% isolates showed extended-spectrum β-lactamase production. Fifty-nine genes reported for antimicrobial resistance and 12 for conferring virulence were targeted. The most prevalent resistance gene for aminoglycosides was aadA (42·3%), for quinolone resistance determining region parE (62·5%), for penicillin's Int1 (62·9%), for chloramphenicol cat3 (66·1%) and for beta-lactams blaTEM -1 (44·4%). Among efflux pump coding genes, armA showed highest (74·2%) prevalence and for virulence, a high prevalence of SopE (89·2%) showed the zoonotic potential of the isolates. The activity of efflux pumps was detected through Ethidium Bromide-agar method. Poultry could act as reservoirs of multidrug resistance Salmonella. We firstly report the prevalence and molecular characterization of virulence/drug resistance in S. Infantis from this region and the results may contribute to designing precisely targeted therapy. This study has also highlighted the possible emergence of S. Infantis with zoonotic potential.

Symbiotic cornucopia of the monophagous planthopper Ommatidiotus dissimilis (Fall\xe9n, 1806) (Hemiptera: Fulgoromorpha: Caliscelidae)

In contrast to Cicadomorpha, in which numerous symbiotic bacteria have been identified and characterized, the symbionts of fulgoromorphans are poorly known. Here, we present the results of histological, ultrastructural, and molecular analyses of the symbiotic system of the planthopper Ommatidiotus dissimilis. Amplification, cloning, and sequencing of bacterial 16S RNA genes have revealed that O. dissimilis is host to five types of bacteria. Apart from bacteria Sulcia and Vidania, which are regarded as ancestral symbionts of Fulgoromorpha, three additional types of bacteria belonging to the genera Sodalis, Wolbachia, and Rickettsia have been detected. Histological and ultrastructural investigations have shown that bacteria Sulcia, Vidania, and Sodalis house separate bacteriocytes, whereas bacteria Wolbachia and Rickettsia are dispersed within various insect tissue. Additionally, bacteria belonging to the genus Vidania occupy the bacteriome localized in the lumen of the hindgut. Both molecular and microscopic analyses have revealed that all the symbionts are transovarially transmitted between generations.

Can soil microbial diversity influence plant metabolites and life history traits of a rhizophagous insect? A demonstration in oilseed rape.

Interactions between plants and phytophagous insects play an important part in shaping the biochemical composition of plants. Reciprocally plant metabolites can influence major life history traits in these insects and largely contribute to their fitness. Plant rhizospheric microorganisms are an important biotic factor modulating plant metabolites and adaptation to stress. While plant-insects or plant-microorganisms interactions and their consequences on the plant metabolite signature are well-documented, the impact of soil microbial communities on plant defenses against phytophagous insects remains poorly known. In this study, we used oilseed rape (Brassica napus) and the cabbage root fly (Delia radicum) as biological models to tackle this question. Even though D. radicum is a belowground herbivore as a larva, its adult life history traits depend on aboveground signals. We therefore tested whether soil microbial diversity influenced emergence rate and fitness but also fly oviposition behavior, and tried to link possible effects to modifications in leaf and root metabolites. Through a removal-recolonization experiment, 3 soil microbial modalities ("high," "medium," "low") were established and assessed through amplicon sequencing of 16S and 18S ribosomal RNA genes. The "medium" modality in the rhizosphere significantly improved insect development traits. Plant-microorganism interactions were marginally associated to modulations of root metabolites profiles, which could partly explain these results. We highlighted the potential role of plant-microbial interaction in plant defenses against Delia radicum. Rhizospheric microbial communities must be taken into account when analyzing plant defenses against herbivores, being either below or aboveground.

Bacteria Associated With Piezodorus guildinii (Hemiptera: Pentatomidae), With Special Reference to Those Transmitted by Feeding.

The redbanded stink bug, Piezodorus guildinii (Westwood) (Hemiptera: Heteroptera: Pentatomidae), is a rapidly growing pest damaging southern US agriculture. Pentatomid stink bugs are known to vector bacterial, fungal, and viral plant diseases. However, bacteria associated with redbanded stink bugs and their vector potential have not yet been assessed. In this study, we 1) cultured and identified bacteria transmitted by feeding of redbanded stink bug and 2) described bacteria from guts of redbanded stink bug individuals using next-generation sequencing of 16S rRNA genes. Nineteen bacteria transmitted by feeding of redbanded stink bug on soybean agar were isolated and identified via Sanger sequencing of near full length 16S RNA genes. The transmitted bacteria belonged to at least a dozen species in eight genera and included potential plant pathogens (Phaseolibacter flectens), plant beneficials (Bacillus atropheus), and possible insect beneficials (Acinetobacter sp. and Citrobacter farmeri). A total of 284,448 reads were captured from Illumina MiSeq sequencing of the uncultured gut bacteria community. Fifty-one putative bacteria species (74% of the estimated total species richness) were identified via matches to NCBI databases. The bacteria metagenome contained potential plant and insect pathogens (Erwinia persicina, E. rhaponici, Brenneria nigrifluens, Ralstonia picketti, and Serratia marcescens) and beneficials (Pantoea dispersa, Klebsiella oxytoca, Clostridium butyricum, and Citrobacter farmeri).

An investigation into blood microbiota and its potential association with Bacterial Chondronecrosis with Osteomyelitis (BCO) in Broilers.

Bacterial chondronecrosis with osteomyelitis (BCO) is a common cause of lameness in commercial broiler chickens worldwide. BCO represents substantial production loss and welfare issues of chickens. The bacterial species or communities underlying BCO pathogenesis still remain to be fully characterized. To gain insights on blood microbiota in broilers and its potential association with BCO, blood samples collected from healthy (n = 240) and lame (n = 12) chickens were analyzed by deep sequencing of 16S RNA genes. The chicken blood microbiota were dominated by Proteobacteria (60.58% ± 0.65) followed by Bactroidetes (13.99% ± 0.29), Firmicutes (11.45% ± 0.51), Actinobacteria (10.21% ± 0.37) and Cyanobacteria (1.96% ± 0.21) that constituted 98.18% (± 0.22) of the whole phyla. The bacterial communities consist of 30–40 OTUs in the blood of broiler chickens, regardless of ages and other environmental or host conditions, and the blood microbiomes of BCO chickens were largely distinct from those of healthy chickens. In addition, Linear discriminant analysis (LDA) effect size (LEfSe) method revealed that Staphylococcus, Granulicatella, and Microbacterium were significantly enriched in BCO chickens as compared to healthy chickens. The results from this study have significant implications in understanding blood microbiota present in broiler chickens and its potential role in BCO pathogenesis.

Effects of varying CO2 flows on bacterial communities in mesocosms created from two soils

Although leakages from carbon capture and storage sites are not expected, it is necessary to be prepared for that possibility and evaluate its risk on soil bacterial communities. Since most studies regarding CO2 effects have been performed in natural sites, the current study was conducted in an experimental site (PISCO2, Spain) in an attempt to consider the short term instead of the long-term effects on bacterial communities, together with the effects of low CO2 below-ground emissions. High-throughput sequencing of 16S RNA genes was used to evaluate the composition and structure of two soils bacterial communities. Overall, communities presented different compositions and structures between soils. The analysis of communities sampled in each soil showed partial shifts in bacterial community composition, but no consistent effects on the richness and diversity. CO2 flux effects on community composition were mainly observed in one soil type, in which the CO2 flux could be considered as one of the factors that contributes to shape bacterial communities, even at low CO2 fluxes and short term emissions.

High-throughput sequencing of 16S RNA genes of soil bacterial communities from a naturally occurring CO2 gas vent

High-throughput sequencing was used to investigate the effects of CO2 emissions on the composition and structure of soil bacterial communities. Soil from an area (La Sima, Campo de Calatrava) where CO2 is released to the atmosphere from a naturally occurring gas vent venting structure. Soil samples were taken was sampled along a CO2 gradient. Bacterial community richness and diversity notably decreased with increasing CO2 flux. Bacterial community composition varied along the CO2 gradient: as CO2 flux increased, the relative abundance of Chloroflexi phylum increased, whereas the relative abundance of Acidobacteria, Verrucomicrobia and Gemmatiomonadetes phyla decreased. Within the Chloroflexi phylum, OTUs related to the genera Thermogemmatispora, Ktedonobacter and Thermomicrobium dominated bacterial communities sampled in sites with the highest CO2 flux. Canonical correspondence analysis (CCA) revealed that differences in bacterial communities were best explained by CO2 flux. The geological storage of CO2 has been proposed as a viable option for reducing anthropogenic CO2 contributions to atmospheric pools in order to mitigate the climatic change. These results are useful to evaluate the risk of potential CO2 leakages on ecosystems associated to geological storage of CO2.

Contrasting the Microbiomes From Healthy Volunteers and Patients With Chronic Rhinosinusitis

Chronic rhinosinusitis (CRS) is the persistent inflammation of the sinus and nasal passages lasting over 3 months. The etiology of CRS is not well understood. To obtain insights into the disease process, we contrasted the microbiome and immune response from patients with CRS and healthy controls. A case vs control design was used. Samples were collected in the operating room in an institutional hospital or clinic. Thirty patients with CRS and 12 healthy controls undergoing surgery were recruited. The microbiome was analyzed by deep sequencing of the bacterial 16S and fungal 18S ribosomal RNA genes. Immune response was measured by quantification of 30 different cytokines by multiplexed enzyme-linked immunosorbent assay, and immune cells in the lavage were identified by flow cytometry. The immune response of peripheral blood leukocytes to the lavage microbiota was assessed by interleukin (IL)-5 enzyme-linked immunospot assay. While quantitative increase in most bacterial and fungal species was observed in patients with CRS relative to controls, the microbiomes of patients with CRS were qualitatively similar to the controls. Because these results indicated that bacteria and fungi are not triggering CRS, we undertook a more detailed characterization of the immune response. Patients with CRS had increased levels of the following cytokines: IL-4, IL-5, IL-8, and IL-13, along with increased levels of eosinophils and basophils in the lavage. Importantly, peripheral blood leukocytes isolated from patients with CRS responded to control lavage samples (ie, to commensals) to produce IL-5. In contrast, the same lavage sample evoked no IL-5 production in leukocytes from healthy controls. These findings support the theory that in some cases CRS results from an immune hyperresponsiveness to commensal organisms.

Microbiome diversity of Aphis glycines with extensive superinfection in native and invasive populations

Associations among insects and microbes can lead to beneficial or parasitic interactions. Using 454 sequencing of 16S RNA genes, we compared microbiome diversity and abundance among field-collected (F) and laboratory-reared (L) populations of the invasive soybean aphid (Aphis glycines), a pest of soybean. Additionally, we screened A. glycines populations from native (Japan, South Korea and China) and invasive regions (North America) to broadly determine the microbiome diversity. Our results suggested that Arsenophonus (relative abundance of 54.6%), Buchnera (38.7%) and Wolbachia (3.7%) were the major bacteria associated with A. glycines. Arsenophonus was the most abundant in F populations but was significantly reduced in L populations; additional bacteria species also had lower relative abundances in L populations. Native and invasive populations were largely similar in bacteria communities and revealed substantial superinfection of Arsenophonus and Wolbachia. The lone exception was a lack of Arsenophonus in A. glycines from Japan. Divergent selection pressures among natural and laboratory populations were inferred as factors driving the differential bacterial communities observed. Our results will allow for improved comparative aphid-symbiont research and broaden our understanding of the interactions among insects, endosymbionts and their environments.

Phylogenetic composition and properties of bacteria coexisting with the fungus Hypholoma fasciculare in decaying wood

White-rot fungi are major degraders of woody materials in terrestrial environments because of their ability to decompose lignin. However, little is known on the possible associations of white-rot fungi with other microorganisms during wood decay. We investigated the numbers, community composition and functional traits of bacteria present in natural wood samples under advanced decay by the white-rot basidiomycete Hypholoma fasciculare. The wood samples contained high numbers of cultivable bacteria (0.2-8 x 10(9) colony forming units (CFU) per g of dry wood). Most cultivable bacteria belonged to Proteobacteria and Acidobacteria (75% and 23% of sequences, respectively). The same phyla were also found to be dominant (59% and 23%, respectively) using a non-culturable quantification technique, namely, direct cloning and sequencing of 16sRNA genes extracted from wood. Bacteria that could be subcultured consisted of acid-tolerant strains that seemed to rely on substrates released by lignocellulolytic enzyme activities of the fungus. There were no indications for antagonism (antibiosis) of the bacteria against the fungus.