Production of organic molecules is largely depending on fossil fuels. A sustainable alternative would be the synthesis of these compounds from CO2 and a cheap energy source, such as H2, CH4, NH3, CO, sulfur compounds or iron(II). Volcanic and geothermal areas are rich in CO2 and reduced inorganic gasses and therefore habitats where novel chemolithoautotrophic microorganisms for the synthesis of organic compounds could be discovered. Here we describe “Candidatus Hydrogenisulfobacillus filiaventi” R50 gen. nov., sp. nov., a thermoacidophilic, autotrophic H2-oxidizing microorganism, that fixed CO2 and excreted no less than 0.54 mol organic carbon per mole fixed CO2. Extensive metabolomics and NMR analyses revealed that Val, Ala and Ile are the most dominant form of excreted organic carbon while the aromatic amino acids Tyr and Phe, and Glu and Lys were present at much lower concentrations. In addition to these proteinogenic amino acids, the excreted carbon consisted of homoserine lactone, homoserine and an unidentified amino acid. The biological role of the excretion remains uncertain. In the laboratory, we noticed the production under high growth rates (0.034 h−1, doubling time of 20 h) in combination with O2-limitation, which will most likely not occur in the natural habitat of this strain. Nevertheless, this large production of extracellular organic molecules from CO2 may open possibilities to use chemolithoautotrophic microorganisms for the sustainable production of important biomolecules.
Spotted fever illness caused by the tick-borne pathogen Rickettsia parkeri has emerged in the Pampa biome in southern Brazil, where the tick Amblyomma tigrinum is implicated as the main vector. Because domestic dogs are commonly parasitized by A. tigrinum, this canid is also a suitable sentinel for R. parkeri-associated spotted fever. Herein, we investigate rickettsial infection in ticks, domestic dogs and small mammals in a natural reserve of the Pampa biome in southern Brazil. The ticks A. tigrinum, Amblyomma aureolatum and Rhipicephalus sanguineus were collected from dogs. Molecular analyses of ticks did not detect R. parkeri; however, at least 34% (21/61) of the A. tigrinum ticks were infected by the non-pathogenic agent ‘Candidatus Rickettsia andeanae’. Serological analyses revealed that only 14% and 3% of 36 dogs and 34 small mammals, respectively, were exposed to rickettsial antigens. These results indicate that the study area is not endemic for R. parkeri rickettsiosis. We tabulated 10 studies that reported rickettsial infection in A. tigrinum populations from South America. There was a strong negative correlation between the infection rates by R. parkeri and ‘Candidatus R. andeanae’ in A. tigrinum populations. We propose that high infection rates by ‘Candidatus R. andeanae’ might promote the exclusion of R. parkeri from A. tigrinum populations. The mechanisms for such exclusion are yet to be elucidated.
Cable bacteria are centimeters-long filamentous bacteria that oxidize sulfide in anoxic sediment layers and reduce oxygen at the oxic-anoxic interface, connecting these reactions via electron transport. The ubiquitous cable bacteria have a major impact on sediment geochemistry and microbial communities. This includes diverse bacteria swimming around cable bacteria as dense flocks in the anoxic zone, where the cable bacteria act as chemotactic attractant. We hypothesized that flocking only appears when cable bacteria are highly abundant and active. We set out to discern the timing and drivers of flocking over 81 days in an enrichment culture of the freshwater cable bacterium Candidatus Electronema aureum GS by measuring sediment microprofiles of pH, oxygen, and electric potential as a proxy of cable bacteria activity. Cable bacterial relative abundance was quantified by 16S rRNA amplicon sequencing, and microscopy observations to determine presence of flocking. Flocking was always observed at some cable bacteria, irrespective of overall cable bacteria rRNA abundance, activity, or sediment pH. Diverse cell morphologies of flockers were observed, suggesting that flocking is not restricted to a specific, single bacterial associate. This, coupled with their consistent presence supports a common mechanism of interaction, likely interspecies electron transfer via electron shuttles. Flocking appears exclusively linked to the electron conducting activity of the individual cable bacteria.
Terrestrial hydrothermal springs and aquifers are excellent sites to study microbial biogeography because of their high physicochemical heterogeneity across relatively limited geographic regions. In this study, we performed 16S rRNA gene sequencing and metagenomic analyses of the microbial diversity of 11 different geothermal aquifers and springs across the tectonically active Biga Peninsula (Turkey). Across geothermal settings ranging in temperature from 43 to 79°C, one of the most highly represented groups in both 16S rRNA gene and metagenomic datasets was affiliated with the uncultivated phylum “Candidatus Bipolaricaulota” (former “Ca. Acetothermia” and OP1 division). The highest relative abundance of “Ca. Bipolaricaulota” was observed in a 68°C geothermal brine sediment, where it dominated the microbial community, representing 91% of all detectable 16S rRNA genes. Correlation analysis of “Ca. Bipolaricaulota” operational taxonomic units (OTUs) with physicochemical parameters indicated that salinity was the strongest environmental factor measured associated with the distribution of this novel group in geothermal fluids. Correspondingly, analysis of 23 metagenome-assembled genomes (MAGs) revealed two distinct groups of “Ca. Bipolaricaulota” MAGs based on the differences in carbon metabolism: one group encoding the bacterial Wood-Ljungdahl pathway (WLP) for H2 dependent CO2 fixation is selected for at lower salinities, and a second heterotrophic clade that lacks the WLP that was selected for under hypersaline conditions in the geothermal brine sediment. In conclusion, our results highlight that the biogeography of “Ca. Bipolaricaulota” taxa is strongly correlated with salinity in hydrothermal ecosystems, which coincides with key differences in carbon acquisition strategies. The exceptionally high relative abundance of apparently heterotrophic representatives of this novel candidate Phylum in geothermal brine sediment observed here may help to guide future enrichment experiments to obtain representatives in pure culture.
Endosymbionts play crucial roles in various physiological activities within insect hosts. The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is an important vector for Candidatus Liberibacter asiaticus (CLas), a fatal pathogenic bacterial agent causing the disease Huanglongbing in the citrus industry. This study combines high-throughput sequencing of 16S ribosomal RNA amplicons to explore how CLas affects the bacterial community in different color morphs (blue, gray), genders, and tissues (cuticle, gut, mycetome, Malpighian tubule, ovary, and testis) of ACP. We found that there was no significant differences in the bacterial community diversity and CLas acquired ratio between the different color morphs and genders of ACP adults. However, acquiring CLas could promote the adult bacterial community’s diversity and richness more than in the uninfected condition. The presence of CLas could increase the Wolbachia and unclassified_Enterobacteriaceae proportions more than in the uninfected condition. The bacterial community diversity in the CLas infected tissues of ovary and cuticle, was lower than the uninfected condition, but the richness of all tissues was not different between the infected and uninfected conditions. CLas could also change the bacterial structure in different tissues and make the bacterial relationship network simpler than it is in an uninfected condition. Furthermore, we used quantitative real-time PCR to assess the dynamic changes of Wolbachia in CLas uninfected and infected color morphs and tissues of ACP. The results showed that Wolbachia titers were significantly higher in CLas infected adults than in uninfected adults. In different tissues, the Wolbachia titers in the testis, ovary, and Malpighian tubule were higher than their uninfected counterparts. Our results provide essential knowledge for understanding the symbionts of the ACP and how CLas affects the bacterial community of the ACP.
The hypervariable genomic regions derived from 35 published
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Las genomes were used to decipher the genetic diversity of
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Las strains and identify 10 new strains with high variations in prophage regions. Characterizing these variations in the
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Las bacteria might provide insight into their evolution and adaptation to host plants and insects in China.
Se identificaron nuevos haplotipos de Diaphorina citri también conocido como el psílido asiático de los cítricos, denominados DcitACC-1, DcitACC-2 y DcitACC-3. Los estudios se basaron en la amplificación de ADN del gen COI mitocondrial y se utilizaron individuos de diferentes zonas citrícolas del país, en algunas zonas productoras del país no se han realizado muestreos con anterioridad, específicamente en los municipios de Acatlán de Pérez, Oaxaca, Misantla y Tantoyuca, Veracruz y Huejutla, Hidalgo, por lo que se procedió a colectar insectos adultos sin distinción de género. El número de individuos de cada sitio colectado dependió de la disponibilidad de insectos en el lugar. Se obtuvieron en total 60 individuos colectados. La amplificación del ADN se realizó con los iniciadores específicos DCITRI COI-L y DCITRI COI-R, el producto de la reacción PCR se secuenció en el Instituto Potosino de Investigación Científica y Tecnológica, AC (IPICYT). Las secuencias obtenidas se compararon con las reportadas en el Genebank y se determinó que existe una línea matriz que corresponde a los haplotipos Dcit-01 y Dcit-04 con número de identificación FJ190300 y FJ190306 (Boykin, 2007). Se obtuvieron 22 secuencias que se analizaron con los programas Oligo analizer y Clustal Omega y se identificaron 11 secuencias iguales a los haplotipos Dcit-01 y Dcit-04. Los resultados mostraron 13 secuencias con diferencias en tres nucleótidos específicos: 61, 253 y 636, mismos que son reportados en este trabajo como nuevos haplotipos.
El objetivo de este trabajo fue identificar en el genoma de Candidatus Liberibacter asiaticus (CLas), proteínas de membrana externa con potencial para el desarrollo y optimización de un método de detección inmunoenzimático. El estudio se realizó durante 2019 y se utilizó el servidor web Predict Protein, así como las bases de datos HhPred/HhSearch y Pfam. Se detectaron 52 proteínas de membrana externa en el genoma completo de CLas, de las cuales, 11 no habían sido caracterizadas previamente. Los análisis predictivos realizados en la proteína B8Y674 generaron ocho posibles epítopos y cuatro de ellos evaluados experimentalmente en células B, mostraron porcentajes de identidad entre 80 a 90%. Se detectó a CLas mediante PCR-punto final a partir del ADN extraído de limón mexicano con síntomas de Huanglongbing utilizando iniciadores diseñados sobre la secuencia del gen Omp que codifica para la proteína B8Y674 y se registró 95% de identidad entre las secuencias generadas y secuencias de CLas previamente reportadas. Los resultados obtenidos nos permiten inferir que la proteína B8Y674 es un candidato potencial para ser utilizada en la detección inmunoenzimática de CLas.