Evaluation of the application potential of Bdellovibrio sp. YBD-1 isolated from Yak faeces.

Studies on Bdellovibrio and like organisms (BALOs), obligate predatory bacteria, have highlighted the possibility of regulating bacteria and biofilms; however, yak-derived BALOs are yet to be reported. We aimed to characterize the BALOs isolated and identified from yak (Bos grunniens) feces and examine application potential. BALOs were isolated from healthy yak fecal samples, with Escherichia coli (ATCC 25922) as prey using the double-layer agar method, identified by transmission electron microscopy (TEM), and the specific 16S rDNA sequencing analysis. Sequencing of the 16S rDNA gene indicated that this isolate was 91% similar to the Bdellovibrio sp. NC01 reference strain and was named YBD-1. Proportion of YBD-1 lysed bacteria is 12/13. The YBD-1 showed best growth at 25-40°C, 0-0.25% (w/v) NaCl, and pH 6.5-7.5. YBD-1 significantly reduced the planktonic cells and biofilms of E.coli in co-culture compared to the E.coli group. Additionally, SEM analysis indicated that YBD-1 significantly reduced biofilm formation in E. coli. Furthermore, quantitative Real Time-polymerase chain reaction (qRT-PCR) showed that the expression of the virulence genes fim and iroN and the genes pgaABC involved in biofilm formation went down significantly. We concluded that YBD-1 may have the potential to control bacterial growth and biofilm-associated bacterial illnesses.

Effects of Acanthamoeba castellanii on the dissolved oxygen and the microbial community under the experimental aquatic model.

Acanthamoeba castellanii is a protist that has a high predation efficiency for bacteria in a number of monoxenic culture experiments. However, the role of A. castellanii in the microbial community is still unknown because of the lack of studies on multiple-species interactions. The aim of this study was to investigate the change of bacterial composition after A. castellanii emerges in a water environment. We added A. castellanii to an environmental water sample and incubated it for two days. Then, we performed 16S ribosomal RNA sequencing techniques to analyze the changes in bacterial composition. In this study, A. castellanii slightly increased the relative abundance of a few opportunistic pathogens, such as Legionella, Roseomonas, and Haemophilus. This result may be related to the training ground hypothesis. On the other hand, the growth of some bacteria was inhibited, such as Cyanobacteria and Firmicutes. Although A. castellanii did not drastically change the whole bacterial community, we surprisingly found the dissolved oxygen concentration was increased after incubation with A. castellanii. We applied environmental water at the laboratory scale to investigate the interactions among A. castellanii, complex microbial communities and the environment. We identified the bacteria that are sensitive to A. castellanii and further found the novel relationship between dissolved oxygen and microbial interaction. Our results helped to clarify the role of A. castellanii in microbial communities.

New 16S rRNA primers to uncover Bdellovibrio and like organisms diversity and abundance.

Appropriate use and specific primers are important in assessing the diversity and abundance of microbial groups of interest. Bdellovibrio and like organisms (BALOs), that refer to obligate Gram-negative bacterial predators of other Gram-negative bacteria, evolved in terms of taxonomy and classification over the past two decades. Hence, some former primers have become inadequate while others are yet to be designed, for both PCR (especially with the advent of NGS) and qPCR approaches. Thus, to study BALOs' abundance and diversity in a variety of aquatic ecosystems, we designed in silico specific primer sets for each BALO genera and tested them in vitro on a variety of cultures and environmental samples. Also, we performed Sanger and Nano Miseq sequencing to reveal the exact degree of specificity of the most promising primers set. Here we report our success in designing specific primers for some BALOs genera, i.e. Bdellovibrio (PCR), Bacteriovorax (qPCR), Peredibacter (PCR).

From the Inside Out: an Epibiotic Bdellovibrio Predator with an Expanded Genomic Complement.

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.

Bdellovibrio and Like Organisms Are Predictors of Microbiome Diversity in Distinct Host Groups.

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.

Diversity, Dynamics, and Distribution of Bdellovibrio and Like Organisms in Perialpine Lakes.

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.

Increasing the autotrophic growth of Chlorella USTB-01 via the control of bacterial contamination by Bdellovibrio USTB-06.

AIMS: (i) To obtain and identify the predatory bacteria for the control of contaminated bacteria and to promote the autotrophic growth of Chlorella USTB-01. (ii) To identify and measure the different cell numbers in microalgal culture using flow cytometer. METHODS AND RESULTS: A predatory bacterial strain was isolated using Escherichia coli BL21 as a sole prey host, which was identified as Bdellovibrio USTB-06 by the analysis of 16S rDNA sequence. A flow cytometer was successfully used to identify and measure the cell numbers of Chlorella USTB-01, the contaminated bacteria and Bdellovibrio USTB-06 simultaneously in the autotrophic culture of Chlorella USTB-01 according to the identification of the different cell sizes. With the addition of Bdellovibrio USTB-06 at initial 104 plaque-forming units per ml, the contaminated bacteria severely decreased by about five counts (in log10  CFU per ml) and the growth of Chlorella USTB-01 was greatly increased by 37·0% compared with those of control respectively. CONCLUSIONS: Bdellovibrio USTB-06 could effectively promote the growth of Chlorella USTB-01 via the killing of the contaminated bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study reveals a good biotechnology method to increase the growth of Chlorella USTB-01 which is very important in the industry of microalgal culture.

Individual Patterns of Complexity in Cystic Fibrosis Lung Microbiota, Including Predator Bacteria, over a 1-Year Period.

Cystic fibrosis (CF) lung microbiota composition has recently been redefined by the application of next-generation sequencing (NGS) tools, identifying, among others, previously undescribed anaerobic and uncultivable bacteria. In the present study, we monitored the fluctuations of this ecosystem in 15 CF patients during a 1-year follow-up period, describing for the first time, as far as we know, the presence of predator bacteria in the CF lung microbiome. In addition, a new computational model was developed to ascertain the hypothetical ecological repercussions of a prey-predator interaction in CF lung microbial communities. Fifteen adult CF patients, stratified according to their pulmonary function into mild (n = 5), moderate (n = 9), and severe (n = 1) disease, were recruited at the CF unit of the Ramón y Cajal University Hospital (Madrid, Spain). Each patient contributed three or four induced sputum samples during a 1-year follow-up period. Lung microbiota composition was determined by both cultivation and NGS techniques and was compared with the patients' clinical variables. Results revealed a particular microbiota composition for each patient that was maintained during the study period, although some fluctuations were detected without any clinical correlation. For the first time, Bdellovibrio and Vampirovibrio predator bacteria were shown in CF lung microbiota and reduced-genome bacterial parasites of the phylum Parcubacteria were also consistently detected. The newly designed computational model allows us to hypothesize that inoculation of predators into the pulmonary microbiome might contribute to the control of chronic colonization by CF pathogens in early colonization stages.IMPORTANCE The application of NGS to sequential samples of CF patients demonstrated the complexity of the organisms present in the lung (156 species) and the constancy of basic individual colonization patterns, although some differences between samples from the same patient were observed, probably related to sampling bias. Bdellovibrio and Vampirovibrio predator bacteria were found for the first time by NGS as part of the CF lung microbiota, although their ecological significance needs to be clarified. The newly designed computational model allows us to hypothesize that inoculation of predators into the lung microbiome can eradicate CF pathogens in early stages of the process. Our data strongly suggest that lower respiratory microbiome fluctuations are not necessarily related to the patient's clinical status.

Isolation of Bdellovibrio sp. from soil samples in Mexico and their potential applications in control of pathogens.

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.

Stenotrophomonas maltophilia biofilm reduction by Bdellovibrio exovorus.

Stenotrophomonas maltophilia, a bacterium ubiquitous in the environment, is also an opportunistic, multidrug-resistant human pathogen that colonizes tissues and medical devices via biofilm formation. We investigated the ability of an isolate from sewage of the bacterial predator Bdellovibrio exovorus to disrupt preformed biofilms of 18 strains of S. maltophilia isolated from patients, hospital sink drains and water fountain drains. B. exovorus FFRS-5 preyed on all S. maltophilia strains in liquid co-cultures and was able to significantly disrupt the biofilms of 15 of the S. maltophilia strains tested, decreasing as much as 76.7% of the biofilm mass. The addition of ciprofloxacin and kanamycin in general reduced S. maltophilia biofilms but less than that of B. exovorus alone. Furthermore, when antibiotics and B. exovorus were used together, B. exovorus was still effective in the presence of ciprofloxacin whereas the addition of kanamycin reduced the effectiveness of B. exovorus. Overall, B. exovorus was able to decrease the mass of preformed biofilms of S. maltophilia in the presence of clinically relevant antibiotics demonstrating that the predator may prove to be a beneficial tool to reduce S. maltophilia environmental or clinically associated biofilms.

Visualizing Bdellovibrio bacteriovorus by Using the tdTomato Fluorescent Protein.

Bdellovibrio bacteriovorus is a Gram-negative bacterium that belongs to the delta subgroup of proteobacteria and is characterized by a predatory life cycle. In recent years, work has highlighted the potential use of this predator to control bacteria and biofilms. Traditionally, the reduction in prey cells was used to monitor predation dynamics. In this study, we introduced pMQ414, a plasmid that expresses the tdTomato fluorescent reporter protein, into a host-independent strain and a host-dependent strain of B. bacteriovorus 109J. The new construct was used to conveniently monitor predator proliferation in real time, in different growth conditions, in the presence of lytic enzymes, and on several prey bacteria, replicating previous studies that used plaque analysis to quantify B. bacteriovorus. The new fluorescent plasmid also enabled us to visualize the predator in liquid cultures, in the context of a biofilm, and in association with human epithelial cells.

Investigation into the Efficacy of Bdellovibrio bacteriovorus as a Novel Preharvest Intervention To Control Escherichia coli O157:H7 and Salmonella in Cattle Using an In Vitro Model.

Cattle are an important reservoir for the foodborne pathogens Salmonella and Escherichia coli O157:H7; they frequently harbor these microorganisms in their digestive tracts and shed them in their feces. Thus, there is potential for contamination of cattle hides and, subsequently, carcasses. Interventions aimed at reducing or eliminating pathogen shedding preharvest will also reduce the likelihood of beef product contamination by these pathogens. Therefore, this study used an in vitro model to evaluate Bdellovibrio bacteriovorus, a gram-negative microorganism that preys upon other gram-negative microorganisms, as a preharvest intervention to control Salmonella and E. coli O157:H7. Rumen fluid and feces were inoculated with pansusceptible or antimicrobial-resistant strains of one pathogen. Control samples were treated with HEPES buffer, whereas experimental samples were exposed to HEPES buffer plus B. bacteriovorus. Salmonella and E. coli O157:H7 populations were quantified at 0, 24, 48, and 72 h. The most-probable-number (MPN) technique, followed by streaking onto xylose lysine Tergitol 4 agar, was used to determine Salmonella populations, whereas spread plating onto sorbitol MacConkey agar supplemented with cefixime and tellurite was employed to enumerate E. coli O157:H7. B. bacteriovorus reduced pansusceptible Salmonella in cattle feces by 2.02 Log MPN/g (P = 0.0005) and antimicrobial-resistant Salmonella by 3.79 (P < 0.0001) and 2.24 (P = 0.0013) Log MPN/g after 24 and 48 h, respectively, in comparison to control samples. Significant reductions were not observed for E. coli O157:H7 in rumen or feces. These data suggest that further investigation into B. bacteriovorus efficacy as a preharvest intervention to control Salmonella in cattle is warranted.

[Diversity of Bdellovibrio-and-like organisms from shrimp mariculture ponds in Zhanjiang].

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.

454 Pyrosequencing reveals diversity of Bdellovibrio and like organisms in fresh and salt water.

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.

To invade or not to invade: two approaches to a prokaryotic predatory life cycle.

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.

Bdellovibrio exovorus sp. nov., a novel predator of Caulobacter crescentus.

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)).

Isolation and classification of Bdellovibrio and like organisms.

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.

Biological characterization of two marine Bdellovibrio-and-like organisms isolated from Daya bay of Shenzhen, China and their application in the elimination of Vibrio parahaemolyticus in oyster.

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.

A microfluidic concentrator array for quantitative predation assays of predatory microbes.

We present a microfabricated concentrator array device that makes it possible to quantify the predation rate of Bdellovibrio bacteriovorus, a predatory microbe, toward its prey, Escherichia coli str. MG1655. The device can accumulate both prey and predator microbes sequentially within a series of concentrator arrays using the motility of the microbes and microfabricated arrowhead-shaped ratchet structures. Since the device can constrain both prey and predator cells within 200 pL chambers at a desired range of cell densities, it was demonstrated that the device cannot only enhance the possibility of studying predation processes/cycles directly at a single cell level but can also quantify the predation rates indirectly by measuring the time-dependent fluorescent intensity signals from the prey. Furthermore, the device can produce a wide range of initial prey to predator density ratios within various concentrator arrays through the use of microfluidic mixer structures on a single array chip, which allows us to study many different conditions with a single set of cultures, and quantitatively characterize the predation behaviour/rate. Lastly, we note that this novel concentrator array device can be a very powerful tool facilitating studies of microbial predations and microbe-microbe interaction and may be broadly used in other microbial biotechnological applications.

Prey bacteria shape the community structure of their predators.

Although predator-prey interactions among higher organisms have been studied extensively, only few examples are known for microbes other than protists and viruses. Among the bacteria, the most studied obligate predators are the Bdellovibrio and like organisms (BALOs) that prey on many other bacteria. In the macroscopical world, both predator and prey influence the population size of the other's community, and may have a role in selection. However, selective pressures among prey and predatory bacteria have been rarely investigated. In this study, Bacteriovorax, a predator within the group of BALOs, in environmental waters were fed two prey bacteria, Vibrio vulnificus and Vibrio parahaemolyticus. The two prey species yielded distinct Bacteriovorax populations, evidence that selective pressures shaped the predator community and diversity. The results of laboratory experiments confirmed the differential predation of Bacteriovorax phylotypes on the two bacteria species. Not only did Bacteriovorax Cluster IX exhibit the versatility to be the exclusive efficient predator on Vibrio vulnificus, thereby, behaving as a specialist, but was also able to prey with similar efficiency on Vibrio parahaemolyticus, indicative of a generalist. Therefore, we proposed a designation of versatilist for this predator. This initiative should provide a basis for further efforts to characterize the predatory patterns of bacterial predators. The results of this study have revealed impacts of the prey on Bacteriovorax predation and in structuring the predator community, and advanced understanding of predation behavior in the microbial world.