RESUMEN
A fundamental question in community ecology is the role of predator-prey interactions in food-web stability and species coexistence. Although microbial microcosms offer powerful systems to investigate it, interrogating the environment is much more arduous. Here, we show in a 1-year survey that the obligate predators Bdellovibrio and like organisms (BALOs) can regulate prey populations, possibly in a density-dependent manner, in the naturally complex, species-rich environments of wastewater treatment plants. Abundant as well as rarer prey populations are affected, leading to an oscillating predatory landscape shifting at various temporal scales in which the total population remains stable. Shifts, along with differential prey range, explain co-existence of the numerous predators through niche partitioning. We validate these sequence-based findings using single-cell sorting combined with fluorescent hybridization and community sequencing. Our approach should be applicable for deciphering community interactions in other systems.
Asunto(s)
Bdellovibrio/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN/métodos , Aguas del Alcantarillado/microbiología , Bacterias/clasificación , Bacterias/genética , Bacterias/crecimiento & desarrollo , Bdellovibrio/clasificación , Bdellovibrio/fisiología , Ecosistema , Cadena Alimentaria , Variación Genética , Filogenia , Dinámica Poblacional , Análisis de la Célula Individual/métodosRESUMEN
Bdellovibrio and like organisms are abundant environmental parasitoids of prokaryotes that show diverse predation strategies. The vast majority of studied Bdellovibrio bacteria and like organisms deploy intraperiplasmic replication inside the prey cell, while few isolates with smaller genomes consume their prey from the outside in an epibiotic manner. The novel parasitoid "Candidatus Bdellovibrio qaytius" was isolated from a eutrophic freshwater pond in British Columbia, where it was a continual part of the microbial community. "Ca Bdellovibrio qaytius" was found to preferentially prey on the betaproteobacterium Paraburkholderia fungorum without entering the periplasm. Despite its epibiotic replication strategy, "Ca Bdellovibrio" encodes a large genomic complement more similar to that of complex periplasmic predators. Functional genomic annotation further revealed several biosynthesis pathways not previously found in epibiotic predators, indicating that "Ca Bdellovibrio" represents an intermediate phenotype and at the same time narrowing down the genomic complement specific to epibiotic predators. In phylogenetic analysis, "Ca Bdellovibrio qaytius" occupies a widely distributed, but poorly characterized, basal cluster within the genus Bdellovibrio This suggests that epibiotic predation might be a common predation type in nature and that epibiotic predation could be the ancestral predation type in the genus.IMPORTANCEBdellovibrio and like organisms are bacteria that prey on other bacteria and are widespread in the environment. Most of the known Bdellovibrio species enter the space between the inner and outer prey membrane, where they consume their prey cells. However, one Bdellovibrio species has been described that consumes its prey from the outside. Here, we describe "Ca Bdellovibrio qaytius," a novel member of the genus Bdellovibrio that also remains outside the prey cell throughout its replication cycle. Unexpectedly, the genome of "Ca Bdellovibrio" is much more similar to the genomes of intracellular predators than to the species with a similar life cycle. Since "Ca Bdellovibrio" is also a basal representative of this genus, we hypothesize that extracellular predation could be the ancestral predation strategy.
Asunto(s)
Bdellovibrio/genética , Bdellovibrio/clasificación , Bdellovibrio/aislamiento & purificación , Bdellovibrio/fisiología , Burkholderiaceae/fisiología , Genoma Bacteriano , Genómica , Filogenia , Estanques/microbiologíaRESUMEN
Biodiversity is generally believed to be a main determinant of ecosystem functioning. This principle also applies to the microbiome and could consequently contribute to host health. According to ecological theory, communities are shaped by top predators whose direct and indirect interactions with community members cause stability and diversity. Bdellovibrio and like organisms (BALOs) are a neglected group of predatory bacteria that feed on Gram-negative bacteria and can thereby influence microbiome composition. We asked whether BALOs can predict biodiversity levels in microbiomes from distinct host groups and environments. We demonstrate that genetic signatures of BALOs are commonly found within the 16S rRNA reads from diverse host taxa. In many cases, their presence, abundance, and especially richness are positively correlated with overall microbiome diversity. Our findings suggest that BALOs can act as drivers of microbial alpha-diversity and should therefore be considered candidates for the restoration of microbiomes and the prevention of dysbiosis.
Asunto(s)
Bdellovibrio/fisiología , Caenorhabditis elegans/microbiología , Drosophila melanogaster/microbiología , Hydra/microbiología , Microbiota , Poríferos/microbiología , Animales , Bdellovibrio/clasificación , Bdellovibrio/genética , Bdellovibrio/aislamiento & purificación , Biodiversidad , ADN Bacteriano/genética , Especificidad del Huésped , Filogenia , Poríferos/clasificación , ARN Ribosómico 16S/genéticaRESUMEN
Defining phenotypic and associated genotypic variation among Bdellovibrio may further our understanding of how this genus attacks and kills different Gram-negative bacteria. We isolated Bdellovibrio sp. NC01 from soil. Analysis of 16S rRNA gene sequences and average amino acid identity showed that NC01 belongs to a different species than the type species bacteriovorus. By clustering amino acid sequences from completely sequenced Bdellovibrio and comparing the resulting orthologue groups to a previously published analysis, we defined a 'core genome' of 778 protein-coding genes and identified four protein-coding genes that appeared to be missing only in NC01. To determine how horizontal gene transfer (HGT) may have impacted NC01 genome evolution, we performed genome-wide comparisons of Bdellovibrio nucleotide sequences, which indicated that eight NC01 genomic regions were likely acquired by HGT. To investigate how genome variation may impact predation, we compared protein-coding gene content between NC01 and the B. bacteriovorus type strain HD100, focusing on genes implicated as important in successful killing of prey. Of these, NC01 is missing ten genes that may play roles in lytic activity during predation. Compared to HD100, NC01 kills fewer tested prey strains and kills Escherichia coli ML35 less efficiently. NC01 causes a smaller log reduction in ML35, after which the prey population recovers and the NC01 population decreases. In addition, NC01 forms turbid plaques on lawns of E. coli ML35, in contrast to clear plaques formed by HD100. Linking phenotypic variation in interactions between Bdellovibrio and Gram-negative bacteria with underlying Bdellovibrio genome variation is valuable for understanding the ecological significance of predatory bacteria and evaluating their effectiveness in clinical applications.
Asunto(s)
Bdellovibrio/fisiología , Genoma Bacteriano/genética , Microbiología del Suelo , Antibiosis/genética , Proteínas Bacterianas/genética , Bdellovibrio/clasificación , Bdellovibrio/genética , Escherichia coli/fisiología , Eliminación de Gen , Transferencia de Gen Horizontal , Bacterias Gramnegativas/fisiología , Viabilidad Microbiana , Fenotipo , Filogenia , ARN Ribosómico 16S/genéticaRESUMEN
Microbes drive a variety of ecosystem processes and services, but many of them remain largely unexplored because of a lack of knowledge on both the diversity and functionality of some potentially crucial microbiological compartments. This is the case with and within the group of bacterial predators collectively known as Bdellovibrio and like organisms (BALOs). Here, we report the abundance, distribution, and diversity of three families of these obligate predatory Gram-negative bacteria in three perialpine lakes (Lakes Annecy, Bourget, and Geneva). The study was conducted at different depths (near-surface versus 45 or 50 m) from August 2015 to January 2016. Using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and cloning-sequencing approaches, we show that the diversity of BALOs is relatively low and very specific to freshwaters or even the lakes themselves. While the Peredibacteraceae family was represented mainly by a single species (Peredibacter starrii), it could represent up to 7% of the total bacterial cell abundances. Comparatively, the abundances of the two other families (Bdellovibrionaceae and Bacteriovoracaceae) were significantly lower. In addition, the distributions in the water column were very different between the three groups, suggesting various life strategies/niches, as follows: Peredibacteraceae dominated near the surface, while Bdellovibrionaceae and Bacteriovoracaceae were more abundant at greater depths. Statistical analyses revealed that BALOs seem mainly to be driven by depth and temperature. Finally, this original study was also the opportunity to design new quantitative PCR (qPCR) primers for Peredibacteraceae quantification.IMPORTANCE This study highlights the abundance, distribution, and diversity of a poorly known microbial compartment in natural aquatic ecosystems, the Bdellovibrio and like organisms (BALOs). These obligate bacterial predators of other bacteria may have an important functional role. This study shows the relative quantitative importance of the three main families of this group, with the design of a new primer pair, and their diversity. While both the diversity and the abundances of these BALOs were globally low, it is noteworthy that the abundance of the Peredibacteraceae could reach important values.
Asunto(s)
Bdellovibrio/aislamiento & purificación , Biodiversidad , Lagos/microbiología , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bdellovibrio/clasificación , Bdellovibrio/genética , ADN Bacteriano/genética , Ecosistema , Filogenia , ARN Ribosómico 16S/genéticaRESUMEN
Obligate predatory bacteria, i.e. bacteria requiring a Gram negative prey cell in order to complete their cell cycle, belong to the polyphyletic group referred to as the Bdellovibrio And Like Organisms (BALO). Predatory interactions between bacteria are complex, yet their dynamics and impact on bacterial communities in the environment are becoming better understood. BALO have unique life cycles: they grow epibiotically with the predator remaining attached to the prey's envelope, dividing in a binary manner or periplasmically, i.e. by penetrating the prey's periplasm to generate a number of progeny cells. The periplasmic life cycle includes unique gene and protein patterns and unique signaling features. These ecological and cellular features, along with applications of the BALO in the medical, agricultural and environmental fields are surveyed.
Asunto(s)
Bdellovibrio/clasificación , Bdellovibrio/patogenicidad , Animales , Antibiosis/fisiología , Proteínas Bacterianas/genética , Bdellovibrio/citología , Ciclo Celular/genética , División Celular , Regulación Bacteriana de la Expresión Génica , HumanosRESUMEN
Acute hepatopancreatic necrosis disease, a severe disease of shrimp, is caused by Vibrio parahaemolyticus (AHPND Vp), a halophilic bacterium harboring a plasmid that contains toxin genes homologous to Photorhabdus insect-related toxins. We obtained 9 isolates of Bdellovibrio and like organisms (BALOs) from water and sediment samples in Thailand. Using 16S rRNA sequencing, all of the organisms were identified as Bacteriovorax spp. and were able to attack all tested AHPND Vp isolates. In addition, their various susceptible hosts, including Gram-positive and Gram-negative bacteria, were observed. The optimal ratio for interaction between the Bacteriovorax isolate BV-A and AHPND Vp was determined to be 1:10. The suitable conditions applied for co-culture between BV-A and AHPND Vp were 30°C, 2% NaCl, and pH 7.6. The capability of BV-A to reduce numbers of AHPND Vp in vitro was observed in co-culture after incubation for 2 d and continued until the end of the incubation period. In vivo, BV-A was able to reduce mortality of shrimp post-larvae infected with AHPND Vp. In addition, BV-A significantly decreased the formation of biofilm by AHPND Vp. These findings provide evidence for using Bacteriovorax as a biocontrol of AHPND Vp in shrimp aquaculture.
Asunto(s)
Bdellovibrio/clasificación , Bdellovibrio/fisiología , Penaeidae/microbiología , Vibrio parahaemolyticus/patogenicidad , Animales , Bdellovibrio/genética , Biopelículas , Interacciones Huésped-Patógeno , Filogenia , ARN Bacteriano/genética , ARN Ribosómico 16S/genéticaRESUMEN
Predatory bacteria are ubiquitously distributed in nature in including in aquatic environments, sewage, intestinal tracts of animals and humans, rhizophere and, soils. However, our understanding of their evolutionary history is limited. Results of recent studies have shown that acquiring novel genes is a major force driving bacterial evolution. Therefore, to gain a better understanding of the impact of gene gain and loss in the evolution of bacterial predators, this study employed comparative genomic approaches to identify core-set gene families and species-specific gene families, and model gene gain and loss events among 11 genomes that represented diverse lineages. In total, 1977 gene families were classified. Of these 509 (pattern 11111111111) were present all of the 11 species. Among the non-core set gene families, 52 were present only in saltwater bacteria predators and had no ortholog in the other genomes. Similarly 109 and 44 were present only in the genomes of Micavibrio spp. and Bdellovibrio spp., respectively. In this study, the gain loss mapping engine GLOOME was selected to analyze and estimate the expectations and probabilities of both gain and loss events in the predatory bacteria. In total, 354 gene families were involved in significant gene gain events, and 407 gene families were classified into gene loss events with high supported value. Moreover, 18 families from the core set gene family were identified as putative genes under positive selection. The results of this study suggest that acquisition of particular genes that encode functional proteins in metabolism and cellular processes and signaling, especially ABC systems, may help bacterial predators adapt to surrounding environmental changes and present different predation strategies for survival in their habitats.
Asunto(s)
Bacterias/genética , Bacterias/patogenicidad , Evolución Molecular , Genes Bacterianos , Alphaproteobacteria/clasificación , Alphaproteobacteria/genética , Alphaproteobacteria/patogenicidad , Animales , Bacterias/clasificación , Bacteroidetes/clasificación , Bacteroidetes/genética , Bacteroidetes/patogenicidad , Bdellovibrio/clasificación , Bdellovibrio/genética , Bdellovibrio/patogenicidad , Mapeo Cromosómico , Deltaproteobacteria/clasificación , Deltaproteobacteria/genética , Deltaproteobacteria/patogenicidad , Ecosistema , Humanos , Familia de Multigenes , Filogenia , Virulencia/genéticaRESUMEN
In this study, two strains of Bdellovibrio were isolated from soil samples using the culture-dependent technique and two members of the family Enterobacteriaceae (Klebsiella sp. and Salmonella sp.) as prey. The Bdellovibrio strains were bacteriolytic, plaque-forming, and highly motile gram-negative bacteria. We identified and confirmed the Bdellovibrio strains using microscopy, PCR amplification, and sequencing of the 16S rRNA gene. They were observed to be different strains based on hit locus and prey range analyses. Here, the first report on Bdellovibrio strains isolated from soil in Mexico corroborates earlier report indicating that populations of Bdellovibrio found in soil are heterogeneous thereby the need to identify the various strains.
Asunto(s)
Bacteriólisis/fisiología , Bdellovibrio/aislamiento & purificación , Agentes de Control Biológico/metabolismo , Klebsiella/crecimiento & desarrollo , Salmonella/crecimiento & desarrollo , Secuencia de Bases , Bdellovibrio/clasificación , Bdellovibrio/genética , ADN Bacteriano/genética , México , Microscopía de Fuerza Atómica , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Microbiología del SueloRESUMEN
Bdellovibrio-and-like organisms (BALOs) are gram-negative, predatory bacteria with wide variations in genome sizes and GC content and ecological habitats. The ATP-binding cassette (ABC) systems have been identified in several prokaryotes, fungi and plants and have a role in transport of materials in and out of cells and in cellular processes. However, knowledge of the ABC systems of BALOs remains obscure. A total of 269 putative ABC proteins were identified in BALOs. The genes encoding these ABC systems occupy nearly 1.3% of the gene content in freshwater Bdellovibrio strains and about 0.7% in their saltwater counterparts. The proteins found belong to 25 ABC system families based on their structural characteristics and functions. Among these, 16 families function as importers, 6 as exporters and 3 are involved in various cellular processes. Eight of these 25 ABC system families were deduced to be the core set of ABC systems conserved in all BALOs. All Bacteriovorax strains have 28 or less ABC systems. On the contrary, the freshwater Bdellovibrio strains have more ABC systems, typically around 51. In the genome of Bdellovibrio exovorus JSS (CP003537.1), 53 putative ABC systems were detected, representing the highest number among all the BALO genomes examined in this study. Unexpected high numbers of ABC systems involved in cellular processes were found in all BALOs. Phylogenetic analysis suggests that the majority of ABC proteins can be assigned into many separate families with high bootstrap supports (>50%). In this study, a general framework of sequence-structure-function connections for the ABC systems in BALOs was revealed providing novel insights for future investigations.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/genética , Proteínas Bacterianas/genética , Bdellovibrio/genética , Deltaproteobacteria/genética , Genoma Bacteriano , Filogenia , Transportadoras de Casetes de Unión a ATP/química , Transportadoras de Casetes de Unión a ATP/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Composición de Base , Bdellovibrio/clasificación , Bdellovibrio/metabolismo , Transporte Biológico , Deltaproteobacteria/clasificación , Deltaproteobacteria/metabolismo , Agua Dulce/microbiología , Expresión Génica , Tamaño del Genoma , SalinidadRESUMEN
Bdellovibrio-and-like organisms (BALOs) are Gram-negative, predatory bacteria that inhabit terrestrial, freshwater and saltwater environments. They have been detected primarily by culture-dependent methods which have limitations. In this study, diversity and community structure of BALOs in freshwater and saltwater samples were characterized by 16S rRNA gene pyrosequencing with specific BALO group primers. Novel Bacteriovorax 16S rDNA sequences were found both in saltwater enrichment cultures and in situ environmental samples, but no new operational taxonomic units were detected in the freshwater samples. The results revealed unexpected diversity of BALOs and advance understanding of the similarities and differences between Bdellovibrio and Bacteriovorax diversity and distribution in the environment.
Asunto(s)
Bdellovibrio/clasificación , Bdellovibrio/aislamiento & purificación , Agua Dulce/microbiología , Variación Genética , Agua de Mar/microbiología , Bdellovibrio/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
OBJECTIVE: Bdellovibrio-and-like organisms (BALOs) are small-sized, parasitic bacteria that rely on other Gram-negative bacteria for survival. Our work aimed to characterize the community diversity of BALOs associated with tropical shrimp ponds. METHODS: We collected water samples from eight shrimp ponds culturing Litopenaeus vannamei, in Zhanjiang, China, and extracted total DNA of the samples. Then the 16S rRNA gene clone libraries were constructed following PCR amplification with BALOs family-specific primers. The community composition and structure of BALOs were further analyzed based on phylogenetic analysis. RESULTS: A total of 726 and 664 valid clones were obtained from the 16S rRNA gene libraries of families Halobacteriovoracaceae and Peredibacteraceae, respectively. Subsequently, they were respectively grouped into 68 and 44 OTUs (operational taxonomic units) at 99.5% sequence similarity, and assigned into 37 and 28 clusters at 97% similarity. For the Halobacteriovoracaceae libraries, most clusters were so far uncultured with the exception of 5 members, which accounting for 43.5% clones. And the cultivable cluster IX and the uncultured cluster B28 were the first and second dominant ones respectively. For the Peredibacteraceae libraries, the cluster pa12 and the only cultivable cluster A3.12 were the first and second dominant ones respectively. Notably, the values of Shannon diversity indices of Halobacteriovoracaceae and Peredibacteraceae tend to be reversed with the change of salinity, but the total BALOs diversities among the eight shrimp ponds were similar. CONCLUSION: Shrimp mariculture ponds in Zhanjinag harbor a high diversity of BALOs comprising of Halobacteriovoracaceae and Peredibacteraceae, and salinity affect their community structure and composition greatly.
Asunto(s)
Bdellovibrio/aislamiento & purificación , Penaeidae/microbiología , Estanques/microbiología , Mariscos/microbiología , Animales , Bdellovibrio/clasificación , Bdellovibrio/genética , Biodiversidad , China , ADN Bacteriano/genética , Datos de Secuencia Molecular , Penaeidae/crecimiento & desarrollo , Filogenia , ARN Ribosómico 16S/genética , Mariscos/análisisRESUMEN
Bdellovibrio and like organisms (BALO) are obligate predators of Gram-negative bacteria, belonging to the α- and δ-proteobacteria. BALO prey using either a periplasmic or an epibiotic predatory strategy, but the genetic background underlying these phenotypes is not known. Here we compare the epibiotic Bdellovibrio exovorus and Micavibrio aeruginosavorus to the periplasmic B. bacteriovorus and Bacteriovorax marinus. Electron microscopy showed that M. aeruginosavorus, but not B. exovorus, can attach to prey cells in a non-polar manner through its longitudinal side. Both these predators were resistant to a surprisingly high number of antibiotic compounds, possibly via 26 and 19 antibiotic-resistance genes, respectively, most of them encoding efflux pumps. Comparative genomic analysis of all the BALOs revealed that epibiotic predators have a much smaller genome (ca. 2.5 Mbp) than the periplasmic predators (ca. 3.5 Mbp). Additionally, periplasmic predators have, on average, 888 more proteins, at least 60% more peptidases, and one more rRNA operon. Fifteen and 219 protein families were specific to the epibiotic and the periplasmic predators, respectively, the latter clearly forming the core of the periplasmic 'predatome', which is upregulated during the growth phase. Metabolic deficiencies of epibiotic genomes include the synthesis of inosine, riboflavin, vitamin B6 and the siderophore aerobactin. The phylogeny of the epibiotic predators suggests that they evolved by convergent evolution, with M. aeruginosavorus originating from a non-predatory ancestor while B. exovorus evolved from periplasmic predators by gene loss.
Asunto(s)
Bdellovibrio/clasificación , Bdellovibrio/fisiología , Evolución Biológica , Bacterias Gramnegativas/fisiología , Proteínas Bacterianas/análisis , Bdellovibrio/citología , Bdellovibrio/genética , Genoma Bacteriano , Filogenia , Proteoma/análisisRESUMEN
Bdellovibrio and like organisms (BALOs) are a group of Gram-negative bacterial predators that are defined as having a periplasmic life cycle, whereby the predator enters into the periplasm of a prey cell. Recently, a predator of Caulobacter crescentus with a novel epibiotic life cycle was identified as a new species - Bdellovibrio exovorus. Therefore, this raises the question as to what determines the type of life cycle of a predator. Six bacterial strains susceptible to predation by B. exovorus JSS were isolated from soil, sewage, and activated sludge. 16S rRNA gene sequence analysis revealed these prey cells to be Acinetobacter johnsonii, Acinetobacter junii, Aeromonas hydrophila, and Delftia acidovorans. The life cycle of B. exovorus was epibiotic on all these prey cells. Environmental samples were enriched with these prey cells; new BALOs were isolated and their life cycle assessed. All new isolates had a periplasmic life cycle. BALOs generally have diverse prey ranges, and thus, not all new prey cells could be used by each new predator. Overall, each prey cell was able to support the growth of predators with either life cycle. Therefore it was confirmed that it is the predator and not the prey that determines the type of life cycle.
Asunto(s)
Bdellovibrio/crecimiento & desarrollo , Cadena Alimentaria , Bacterias Gramnegativas , Microbiología del Suelo , Bdellovibrio/clasificación , Bdellovibrio/genética , Bdellovibrio/aislamiento & purificación , Caulobacter , Medios de Cultivo , Datos de Secuencia MolecularRESUMEN
The life cycle, prey range and taxonomic status of a Bdellovibrio-like organism, strain JSS(T), were studied. Strain JSS(T) was isolated from sewage in London, Ontario, Canada, in enrichment culture with Caulobacter crescentus prey cells. During predation, this strain remained attached to the outside of a stalked C. crescentus cell. No periplasmic growth stage was observed and no bdelloplast was formed. The stalked cells of C. crescentus retained their shape and, after predation, were devoid of cytoplasmic content, as shown by transmission electron microscopy. A periplasmic growth stage has been a definitive character in the description of members of the genera Bdellovibrio, Bacteriovorax, Bacteriolyticum and Peredibacter. This is the first description of an epibiotic predator in this group of prokaryotic predators. The G+C content of the genomic DNA of strain JSS(T) was 46.1 mol%. 16S rRNA gene sequence analysis showed that this strain was related to Bdellovibrio bacteriovorus strains HD100(T), 109J, 114 and 127 (90-93 % similarity). Phylogenetic analysis based on 16S rRNA gene sequences grouped strain JSS(T) with the Bdellovibrio cluster, but at a distance from other Bdellovibrio isolates. On the basis of features of the life cycle and phylogenetic data, it was concluded that strain JSS(T) merits classification as the type strain of a novel species, for which the name Bdellovibrio exovorus sp. nov. is proposed (type strain JSS(T) =ATCC BAA-2330(T) = DSM 25223(T)).
Asunto(s)
Bdellovibrio/clasificación , Filogenia , Aguas del Alcantarillado/microbiología , Técnicas de Tipificación Bacteriana , Composición de Base , Bdellovibrio/genética , Bdellovibrio/aislamiento & purificación , Caulobacter crescentus/crecimiento & desarrollo , ADN Bacteriano/genética , Datos de Secuencia Molecular , Ontario , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
Bacteriovorax marinus SJ is a predatory delta-proteobacterium isolated from a marine environment. The genome sequence of this strain provides an interesting contrast to that of the terrestrial predatory bacterium Bdellovibrio bacteriovorus HD100. Based on their predatory lifestyle, Bacteriovorax were originally designated as members of the genus Bdellovibrio but subsequently were re-assigned to a new genus and family based on genetic and phenotypic differences. B. marinus attaches to gram-negative bacteria, penetrates through the cell wall to form a bdelloplast, in which it replicates, as shown using microscopy. Bacteriovorax is distinct, as it shares only 30% of its gene products with its closest sequenced relatives. Remarkably, 34% of predicted genes over 500 nt in length were completely unique with no significant matches in the databases. As expected, Bacteriovorax shares several characteristic loci with the other delta-proteobacteria. A geneset shared between Bacteriovorax and Bdellovibrio that is not conserved among other delta-proteobacteria such as Myxobacteria (which destroy prey bacteria externally via lysis), or the non-predatory Desulfo-bacteria and Geobacter species was identified. These 291 gene orthologues common to both Bacteriovorax and Bdellovibrio may be the key indicators of host-interaction predatory-specific processes required for prey entry. The locus from Bdellovibrio bacteriovorus is implicated in the switch from predatory to prey/host-independent growth. Although the locus is conserved in B. marinus, the sequence has only limited similarity. The results of this study advance understanding of both the similarities and differences between Bdellovibrio and Bacteriovorax and confirm the distant relationship between the two and their separation into different families.
Asunto(s)
Bdellovibrio/genética , Deltaproteobacteria/genética , Agua de Mar/microbiología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Bdellovibrio/clasificación , Bdellovibrio/fisiología , Deltaproteobacteria/clasificación , Deltaproteobacteria/fisiología , Cadena Alimentaria , Regulación Bacteriana de la Expresión Génica , Genoma Bacteriano , Datos de Secuencia Molecular , Filogenia , PlásmidosRESUMEN
Bdellovibrio and like organisms (BALOs) are obligate predators of Gram-negative bacteria. BALOs are isolated as plaques growing at the expense of their prey and are cultivated as two-member cultures. The growth cycle is composed of an extracellular attack phase and an intraperiplasmic elongation and replication phase. However, there are methods for obtaining host-independent (HI) mutants that grow without prey on rich media. BALOs are commonly found in the environment but generally constitute small populations; therefore, their isolation may require enrichment steps. Contamination by other bacteria during isolation necessitates efficient separation between the smaller BALO cells from the majority of larger bacteria. BALOs can also be directly detected and quantified in environmental samples using specific PCR. Synchronous cultures of both wild-type and HI derivatives can be obtained to study the different growth phases. These can be further separated by centrifugation. Classification is based on 16S rDNA analysis. Protocols relevant to these aspects of BALO detection, isolation, growth, classification, and quantitation are presented in this unit.
Asunto(s)
Técnicas Bacteriológicas/métodos , Bdellovibrio/clasificación , Bdellovibrio/aislamiento & purificación , Centrifugación/métodos , Recuento de Colonia Microbiana/métodos , Filtración/métodos , Bdellovibrio/genética , Bdellovibrio/crecimiento & desarrollo , Reacción en Cadena de la Polimerasa/métodos , Ribotipificación , Microbiología del Suelo , Microbiología del AguaRESUMEN
Recent studies have revealed that the use of bdellovibrios is an alternative to control bacteriosis. However, no bdellovibrios are available against Aeromonas hydrophila infections in sturgeons. In the present study, a potential Bdellovibrio strain F16 was isolated from sturgeon gut samples, using a sturgeon-pathogenic A. hydrophila as the prey bacterium. It was initially identified as a Bdellovibrio strain using morphological characteristics and specific PCR amplification, and confirmed to be Bdellovibrio sp. strain ETB (GenBank Accession No. DQ302728) and Bdellovibrio bacteriovorus strain SRA9 (GenBank Accession No. AF263833) by phylogenetic analysis. In addition, it was shown to be safe for mammalians and sturgeons, had a wide prey range, and exhibited significant bacteriolytic effects on the pathogenic A. hydrophila. To the best of our knowledge, this is the first report on a promising gut Bdellovibrio strain against pathogenic A. hydrophila.
Asunto(s)
Aeromonas hydrophila/fisiología , Antibiosis , Bdellovibrio/fisiología , Agentes de Control Biológico , Aeromonas hydrophila/clasificación , Aeromonas hydrophila/genética , Aeromonas hydrophila/patogenicidad , Animales , Bacterias/genética , Bdellovibrio/clasificación , Bdellovibrio/genética , Enfermedades de los Peces/microbiología , Infecciones por Bacterias Gramnegativas/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Filogenia , Reacción en Cadena de la PolimerasaRESUMEN
Bdellovibrio-and-like organisms (BALOs) are a group of highly motile delta-proteobacteria that prey on other gram-negative bacteria. However, nothing is known of the application potential of marine BALOs in safeguarding seafood safety. Here, biological characterization of two marine BALOs strains and their application in the elimination of Vibrio parahaemolyticus in oyster (Crassostrea ariakensis) at the laboratory scale were investigated. BALOs strains BDH12 and BDHSH06 were isolated from sediment of Daya bay in Shenzhen of China, with Shewanella putrefaciens strain 12 and V. parahaemolyticus strain SH06 as preys, respectively, when using double layer agar technique. They were identified as BALOs morphologically by transmission electron microscopy, while partial 16S rDNA sequencing analysis revealed that they showed no close relationships with members of the known genera Bdellovibrio, Bacteriolyticum, Bacteriovorax, or Peredibacter. Biological characterizations revealed that both strains had the optimal pH, salinity and temperature at 7.2, 3% and 30 °C, correspondingly. They could not utilize autoclaved, dead cells as hosts. Prey range analysis revealed that individually, BDH12 and BDHSH06 lysed 82.5% (47 strains) and 84.2% (48 strains) of the total 57 preys tested respectively. In combination, they lysed 98.2% (56 of 57) strains. All strains of V. parahaemolyticus, Vibrio cholerae and Vibrio alginolyticus tested could be lysed by both strains. A 7-day laboratory-scale V. parahaemolyticus elimination experiment in oyster showed that in the control, the cell counts of total vibrios and V. parahaemolyticus strain Vp plus in water and in oyster intestines were on the rise, whereas in the BALOs treated groups, their numbers were down from 8.09±0.05 log CFU/ml and 8.02±0.04 log CFU/ml to 2.39±0.01 log CFU/ml and 2.33±0.01 log CFU/ml, respectively. The same patterns could also be observed in oyster intestines. Results of this study indicate the feasibility of using BALOs to biologically control or even eliminate V. parahaemolyticus in seafood oyster.
Asunto(s)
Bdellovibrio/aislamiento & purificación , Agentes de Control Biológico , Ostreidae/microbiología , Vibrio parahaemolyticus/crecimiento & desarrollo , Animales , Bdellovibrio/clasificación , Bdellovibrio/genética , Bdellovibrio/crecimiento & desarrollo , China , Recuento de Colonia Microbiana , ADN Bacteriano/genética , Concentración de Iones de Hidrógeno , Filogenia , ARN Ribosómico 16S/genética , Salinidad , Alimentos Marinos/microbiología , Agua de Mar/microbiología , Análisis de Secuencia de ADN , TemperaturaRESUMEN
Bdellovibrio bacteriovorus is a famously fast, flagellate predatory bacterium, preying upon Gram-negative bacteria in liquids; how it interacts with prey on surfaces such as in medical biofilms is unknown. Here we report that Bdellovibrio bacteria "scout" for prey bacteria on solid surfaces, using slow gliding motility that is present in flagellum-negative and pilus-negative strains.