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Although nitrogen removal by partial nitritation and anammox is more cost-effective than conventional nitrification and denitrification, one downside is the production and accumulation of nitrous oxide (N2O). The potential exploitation of N2O-reducing bacteria, which are resident members of anammox microbial communities, for N2O mitigation would require more knowledge of their ecophysiology. This study investigated the phylogeny of resident N2O-reducing bacteria in an anammox microbial community and quantified individually the processes of N2O production and N2O consumption. An up-flow column-bed anammox reactor, fed with NH4+ and NO2- and devoid of oxygen, emitted N2O at an average conversion ratio (produced N2O: influent nitrogen) of 0.284%. Transcriptionally active and highly abundant nosZ genes in the reactor biomass belonged to the Burkholderiaceae (clade I type) and Chloroflexus genera (clade II type). Meanwhile, less abundant but actively transcribing nosZ strains were detected in the genera Rhodoferax, Azospirillum, Lautropia, and Bdellovibrio and likely act as an N2O sink. A novel 15N tracer method was adapted to individually quantify N2O production and N2O consumption rates. The estimated true N2O production rate and true N2O consumption rate were 3.98 ± 0.15 and 3.03 ± 0.18 mgN·gVSS-1·day-1, respectively. The N2O consumption rate could be increased by 51% (4.57 ± 0.51 mgN·gVSS-1·day-1) with elevated N2O concentrations but kept comparable irrespective of the presence or absence of NO2-. Collectively, the approach allowed the quantification of N2O-reducing activity and the identification of transcriptionally active N2O reducers that may constitute as an N2O sink in anammox-based processes.

The small size of bacterial cells necessitates rapid adaption to sudden environmental changes. In Bdellovibrio bacteriovorus, an obligate predator of bacteria common in oligotrophic environments, the non-replicative, highly motile attack phase (AP) cell must invade a prey to ensure replication. AP cells swim fast and respire at high rates, rapidly consuming their own contents. How the predator survives in the absence of prey is unknown. We show that starvation for prey significantly alters swimming patterns and causes exponential decay in prey-searching cells over hours, until population-wide swim-arrest. Swim-arrest is accompanied by changes in energy metabolism, enabling rapid swim-reactivation upon introduction of prey or nutrients, and a sweeping change in gene expression and gene regulation that largely differs from those of the paradigmatic stationary phase. Swim-arrest is costly as it imposes a fitness penalty in the form of delayed growth. We track the control of the swim arrest-reactivation process to cyclic-di-GMP (CdG) effectors, including two motility brakes. CRISPRi transcriptional inactivation, and in situ localization of the brakes to the cell pole, demonstrated their essential role for effective survival under prey-induced starvation. Thus, obligate predators evolved a unique CdG-controlled survival strategy, enabling them to sustain their uncommon lifestyle under fluctuating prey supply.

The effects on haematological parameters of Sprague-Dawley rats infected (challenged) by the injection of pathogenic Pasteurella multocida and the administration of Bdellovibrio bacteriovorus (ATCC™ 1534) as therapy, occasioned by the consideration of its use as an alternative to antibiotics, due to the high rate of bacterial resistance to current clinically used antibiotics was investigated. A total of 60 rats, divided into 5 groups (4 experimental groups and 1 control group) of 12 rats each were used in this study. The first group of 12 rats were injected subcutaneously with one millilitre of 1 x 108/ml B. bacteriovorus (ATCC™ 1534). The second group of 12 rats were injected with one millilitre 108/ml of P. multocida in saline. One millilitre each of 108/ml of both P. multocida and B. bacteriovorus (ATCC™ 1534) were injected into another set of 12 rats in the third group. The third group of 12 rats were injected once intra-muscularly, in the hind flank muscle, with 2 mg/kg of Ketamine Hydrochloride. And lastly, one set of 12 rats were not injected with any bacteria served as control. In all cases, observed haematological data were analysed from the experimental rats after 168 hours (except rats which were injected with 2mg/kg ketamine hydrochloride, used as anaesthetic, (at sacrifice), and compared with the haematological profiles of the 12 control rats. Results shows a reduction of mortality from 9 to 1 (88.8%) of rats challenged with P. multocida over those inoculated with B. bacteriovorus and P. multocida was observed. WBC counts were higher (4.12 x 103/µL) in B. bacteriovorus and P. multocida over WBC counts in control rats which served as WBC reference values. Though, not statistically significant (ANOVA = p >0.05). In a similar comparison, RBC counts (6.5 x 103/µL) were lower than observed in control rats, while platelet counts (1138 x 103/µL) were higher than values in controls, however, this was statistically significant. Moreover, haemoglobin concentrations were lower (11.7 g/dL) than in control rats. Though there were slight variations in haematological profiles from reference values, it was concluded that B. bacteriovorus seems to have no life-threatening effect on haematology of rats. However, evaluations such as observed platelet increases on inoculations of B. bacteriovorus, need to be addressed before the promise of its in vivo use in controlling Gram-negative infection in animals and humans can be tapped. Sar, T. T. | Department of Biological Sciences, University of Agriculture, Makurdi, Benue State, Nigeria

International audience; Almost sixty years ago, Bdellovibrio and like organisms (BALOs) were discovered as the first obligate bacterial predators of other bacteria known to science. Since then, they were shown to be diverse and ubiquitous in the environment, and to bear astonishing ecological, physiological, and metabolic capabilities. The last decade has seen important strides made in understanding the mechanistic basis of their life cycle, the dynamics of their interactions with prey, along with significant developments towards their use in medicine, agriculture, and industry. This review details these achievements, identify current understanding and knowledge gaps to encourage and guide future BALO research.

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.

Bdellovibrio and like organisms (BALOs) are Gram-negative obligate predators of other bacteria in a range of environments. The recent discovery of BALOs in the circulatory system of cultured spiny lobster P. ornatus warrants more investigation. We used a combination of co-culture agar and broth assays and transmission electron microscopy to show a Halobacteriovorax sp. strain Hbv preyed upon the model prey bacterium Vibrio sp. strain Vib. The haemolymph microbiome of juvenile P. ornatus was characterised following injection of phosphate buffered saline (control) or prey and/or predator bacteria for 3 d. The predator Hbv had no effect on survival compared to the control after 3 d. However, when compared to the prey only treatment group, lobsters injected with both prey and predator showed significantly lower abundance of genus Vibrio in the haemolymph bacterial community composition. This study indicates that predatory bacteria are not pathogenic and may assist in controlling microbial population growth in the haemolymph of lobsters.

Bdellovibrio and like organisms (BALOs) prey on Gram-negative bacteria in the planktonic phase as well as in biofilms, with the ability to reduce prey populations by orders of magnitude. During the last few years, evidence has mounted for a significant ecological role for BALOs, with important implications for our understanding of microbial community dynamics as well as for applications against pathogens, including drug-resistant pathogens, in medicine, agriculture and aquaculture, and in industrial settings for various uses. However, our understanding of biofilm predation by BALOs is still very fragmentary, including gaps in their effect on biofilm structure, on prey resistance, and on evolutionary outcomes of both predators and prey. Furthermore, their impact on biofilms has been shown to reach beyond predation, as they are reported to reduce biofilm structures of non-prey cells (including Gram-positive bacteria). Here, we review the available literature on BALOs in biofilms, extending known aspects to potential mechanisms employed by the predators to grow in biofilms. Within that context, we discuss the potential ecological significance and potential future utilization of the predatory and enzymatic possibilities offered by BALOs in medical, agricultural and environmental applications.

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., Studying the role of predator–prey interactions in food-web stability and species coexistence in the environment is arduous. Here, Cohen et al. use a combination of community and single-cell analyses to show that bacterial predators can regulate prey populations in the species-rich environments of wastewater treatment plants.

It was previously demonstrated that predatory bacteria are able to efficiently eliminate Gram-negative pathogens including antibiotic-resistant and biofilm-associated bacteria. In this proof-of-concept study we evaluated whether two species of predatory bacteria, Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus, were able to alter the survival of Gram-negative pathogens on the ocular surface. Clinical keratitis isolates of Pseudomonas aeruginosa (strain PAC) and Serratia marcescens (strain K904) were applied to the ocular surface of NZW rabbits followed by application of predatory bacteria. At time intervals, surviving pathogenic bacteria were enumerated. In addition, B. bacteriovorus and S. marcescens were applied to porcine organ culture corneas under contact lenses, and the ocular surface was examined by scanning electron microscopy. The ocular surface epithelial layer of porcine corneas exposed to S. marcescens, but not B. bacteriovorus was damaged. Using this model, neither pathogen could survive on the rabbit ocular surface for longer than 24 h. M. aeruginosavorus correlated with a more rapid clearance of P. aeruginosa but not S. marcescens from rabbit eyes. This study supports previous evidence that predatory bacteria are well tolerated by the cornea, but suggest that predatory bacteria do not considerably change the ability of the ocular surface to clear the tested Gram-negative bacterial pathogens from the ocular surface.

As microbiologists and public health officials scramble for weapons to combat antibiotic resistance, they may end up including an unlikely ally in their arsenal: other bacteria. The prime candidate right now is a predatory bacterium known as Bdellovibrio bacteriovorus . Researchers have started to evaluate predatory bacteria such as Bdellovibrio —shown here in a false-color transmission electron microscopy image at 50,000× magnification—as a means of treating intractable, antibiotic-resistant infections. Image credit: Science Source/ALFRED PASIEKA. Found in soils and water globally, B. bacteriovorus are free-living, harmless bacteria—harmless to humans, at least. In the microbial world, they’re voracious predators of gram-negative bacteria, a group that includes Salmonella , Escherichia coli , and several other pathogens. Bdellovibrio punch a hole in the membranes of their prey, enter and consume their contents, then burst out again to find their next meal. “They’re a very efficient killing machine,” says Daniel Kadouri, an associate professor at the Rutgers School of Dental Medicine in Newark, NJ. “They act kind of like the creature in the movie Alien .” Researchers are now beginning to evaluate Bdellovibrio and other similar predatory bacteria as a route to treating intractable, antibiotic-resistant infections. They’re starting to learn how these predators work in different organs and against different pathogens. As they examine various infection scenarios, they are also discovering how these microbes work in concert with the native microbiome and immune cells. Pills full of predatory bacteria won’t replace antibiotics at pharmacies anytime soon. But studying these microbes could help prepare for a future when many antibiotics fail to treat multi-drug-resistant infections. “Right now, the idea that a single injection of live bacteria could be used as therapy seems quite extreme to people,” says microbiologist Elizabeth Sockett of the University of Nottingham in the United Kingdom. “But as we run out of other …

The use of bdellovibrios has been regarded as an alternative to control multidrug-resistant pathogens and fish bacteriosis. However, scarce information is available on the potential of bdellovibrios in the presence of copper sulfate, which is an algicide widely used to treat cyanobacterial blooms in aquaculture. In the present study, the effects of copper sulfate at sublethal and lethal levels (0.1 and 1.0 mg·L−1) on Bdellovibrio sp. strain BDF-H16 were evaluated. The growth of Bdellovibrio sp. strain BDF-H16 was significantly promoted by both concentrations of copper sulfate, but less so by the lethal concentration. The bacteriolysis of gibel carp-pathogenic Aeromonas hydrophila by Bdellovibrio sp. strain BDF-H16 was also stimulated by copper sulfate in both solid and liquid media. However, Bdellovibrio sp. strain BDF-H16 with 0.1 mg·L−1 copper sulfate clearly inhibited infection of gibel carps by A. hydrophila better than Bdellovibrio sp. strain BDF-H16 with 1.0 mg·L−1 copper sulfate did.

AIMS The Mediterranean fruit fly (the medfly) causes major losses of agricultural fruits. Its microbiome is mainly composed of various Enterobacteriaceae that contribute to nutrient acquisition and are associated with the fly's development. Moreover, the performance of males produced by the sterile insect technique is improved by providing mass-reared insects with specific gut bacteria. Bdellovibrio and like organisms (BALOs) are obligate predators of Gram-negative bacteria that efficiently preys upon diverse Enterobacteriaceae, making it a potential disruptor of the fly's microbiome. We hypothesized that the fly's microbiome can be targeted to control the insect. METHODS AND RESULTS Inoculation of B. bacteriovorus as free-swimming or encapsulated cells into gut extracts significantly reduced gut bacterial abundance, sustaining predator survival. Similar treatments applied to adult flies showed that the predators also survived in the gut environment. While addition of the predators did not affect total gut bacterial abundance and end-point fly mortality, a shift in the gut community structure, measured by high-throughput community sequencing was observed. CONCLUSIONS The bacterial predator of bacteria B. bacteriovorus can prey and survive in vivo in the medfly gut. SIGNIFICANCE AND IMPACT OF THE STUDY This study establishes the potential of BALOs to affect the microbiome of insect hosts.

Bdellovibrio bacteriovorus is an environmentally-ubiquitous bacterium that uses unique adaptations to kill other bacteria. The best-characterized strain, HD100, has a multistage lifestyle, with both a free-living attack phase and an intraperiplasmic growth and division phase inside the prey cell. Advances in understanding the basic biology and regulation of predation processes are paving the way for future potential therapeutic and bioremediation applications of this unusual bacterium.

Recent studies investigating Bdellovibrio spp. have found that although this predator predominantly preys on Gram-negative organisms, under certain conditions (nutrient/prey limitation), it will adapt to survive and grow axenically (without prey) or in the presence of Gram-positive bacterial prey. These advances in the understanding of predatory bacteria have stimulated a renewed interest in these organisms and the potential applications of Bdellovibrio spp. to the benefit of society. Early studies primarily focused on the application of predatory bacteria as "live antibiotics" in the medical field, probiotics in aquaculture and veterinary medicine and their use in agriculture. Additionally, studies have investigated their prevalence in wastewater and environmental sources. However, comprehending that Bdellovibrio spp. may also prey on and target Gram-positive organisms, implies that these predators could specifically be applied for the bioremediation or removal of mixed bacterial communities. Recent studies have also indicated that Bdellovibrio spp. may be useful in controlling food spoilage organisms and subsequently decrease our reliance on food additives. This review will thus highlight recent developments in understanding Bdellovibrio spp. predation strategies and focus on potential new applications of these organisms for water treatment, food preservation, enhancement of industrial processes, and in combination therapies with bacteriophages and/or antibiotics to combat multi-drug resistant organisms.

Glycolysis and gluconeogenesis are central pathways of metabolism across all domains of life. A prominent enzyme in these pathways is phosphoglucose isomerase (PGI) which mediates the interconversion of glucose-6-phosphate and fructose-6-phosphate (F6P). The predatory bacterium Bdellovibrio bacteriovorus leads a complex lifecycle, switching between intraperiplasmic replicative and extracellular “hunter” attack-phase stages. Passage through this complex lifecycle involves different metabolic states. Here we present the unliganded and substrate bound structures of the Bdellovibrio bacteriovorus PGI, solved to 1.74Å and 1.67 Å, respectively. These structures reveal that an induced-fit conformational change within the active site is not a pre-requisite for the binding of substrates in some PGIs. Crucially, we suggest a phenylalanine residue, conserved across most PGI enzymes but substituted for a glycine in Bdellovibrio and other select organisms, is central to the induced-fit mode of substrate recognition for PGIs. This enzyme also represents the smallest conventional PGI characterised to date and likely represents the minimal requirements for a functional PGI.

Trophic interactions are crucial for carbon cycling in food webs. Traditionally, eukaryotic micropredators are considered the major micropredators of bacteria in soils, although bacteria like myxobacteria and Bdellovibrio are also known bacterivores. Until recently, it was impossible to assess the abundance of prokaryotes and eukaryotes in soil food webs simultaneously. Using metatranscriptomic three-domain community profiling we identified pro- and eukaryotic micropredators in 11 European mineral and organic soils from different climes. Myxobacteria comprised 1.5–9.7% of all obtained SSU rRNA transcripts and more than 60% of all identified potential bacterivores in most soils. The name-giving and well-characterized predatory bacteria affiliated with the Myxococcaceae were barely present, while Haliangiaceae and Polyangiaceae dominated. In predation assays, representatives of the latter showed prey spectra as broad as the Myxococcaceae. 18S rRNA transcripts from eukaryotic micropredators, like amoeba and nematodes, were generally less abundant than myxobacterial 16S rRNA transcripts, especially in mineral soils. Although SSU rRNA does not directly reflect organismic abundance, our findings indicate that myxobacteria could be keystone taxa in the soil microbial food web, with potential impact on prokaryotic community composition. Further, they suggest an overlooked, yet ecologically relevant food web module, independent of eukaryotic micropredators and subject to separate environmental and evolutionary pressures.

Bdellovibrio and like organisms (BALOs) are obligate predatory bacteria commonly encountered in the environment. In dual predator–prey cultures, prey accessibility ensures optimal feeding and replication and rapid BALO population growth. However, the environmental prey landscape is complex, as it also incorporates non-prey cells and other particles. These may act as decoys, generating unproductive encounters which in turn may affect both predator and prey population dynamics. In this study, we hypothesized that increasing decoy:prey ratios would bring about increasing costs on the predator's reproductive fitness. We also tested the hypothesis that different BALOs and decoys would have different effects. To this end, we constructed prey landscapes including periplasmic or epibiotic predators including two types of decoy under a large range of initial decoy:prey ratio, and mixed cultures containing multiple predators and prey. We show that as decoy:prey ratios increase, the maximal predator population sizes is reduced and the time to reach it significantly increases. We found that BALOs spent less time handling non-prey (including superinfection-immune invaded prey) than prey cells, and did not differentiate between efficient and less efficient prey. This may explain why in multiple predator and prey cultures, less preferred prey appear to act as decoy.

In this study, two bacterial strains, IRP7 and IRP8, were selected to induce resistance against pine wilt disease (PWD). Foliar application with these strains to nematode-inoculated pine seedlings significantly reduced PWD severity. The effect of nematode inoculation and bacterial treatment on the rhizosphere bacterial community was investigated. The results indicated that the rhizosphere of nematode-inoculated seedlings contained a lower relative abundance of beneficial microbes such as Paraburkholderia, Bradyrhizobium, Rhizobacter, Lysobacter, and Caballeronia. Bacterial treatment resulted in significant changes in the microbes that were represented in relatively low relative abundance. Treatment with IRP7 resulted in an increase in the relative abundance of Nitrospirillum, Bacillus, and Luteibacter, which might be useful for protection against infection. Treatment with IRP8 resulted in an increase in the relative abundance of obligate bacterial predators of the Bdellovibrio genus that were previously shown to control several bacterial phytopathogens and may have a role in the management of nematode-carried bacteria. The selected bacteria were identified as Pseudomonas koreensis IRP7 and Lysobacter enzymogenes IRP8 and are suggested as a potential treatment for induced resistance against PWD. To our knowledge, this is the first report on the effect of foliar treatment with resistance-inducing bacteria on the rhizosphere microbiota.

Antimicrobial resistance (AMR) is a global health and economic crisis. With too few antibiotics in development to meet current and anticipated needs, there is a critical need for new therapies to treat Gram-negative infections. One potential approach is the use of living predatory bacteria, such as Bdellovibrio bacteriovorus (small Gram-negative bacteria that naturally invade and kill Gram-negative pathogens of humans, animals and plants). Moving toward the use of Bdellovibrio as a ‘living antibiotic’ demands the investigation and characterization of these bacterial predators in biologically relevant systems. We review the fundamental science supporting the feasibility of predatory bacteria as alternatives to antibiotics.

Bdellovibrio bacteriovorus, a Gram-negative predatory bacterium belonging to the Bdellovibrio and like organisms (BALOs), predates on Gram-negative bacteria. BALO strains differ in prey range but so far, the genetic basis of resistance against BALO predation is hardly understood. We developed a loss-of-function approach to screen for sensitive mutants in a library of strain M6, a predation-resistant strain of the plant pathogen Acidovorax citrulli. The screen is based on tracking the growth of a B. bacteriovorus strain expressing the fluorescent reporter Tdtomato in mutant pools to reveal predation-sensitive variants. Two independent loci were identified in mutant strains exhibiting significant levels of susceptibility to the predator. Genes in the two loci were analysed using both protein sequence homology and protein structure modeling. Both were secretion-related proteins and thus associated to the bacterial cell wall. Successful complementation of gspK, a gene encoding for a minor pseudopilin protein confirmed the involvement of the type II secretion system in A. citrulli M6 resistance. This proof of concept study shows that our approach can identify key elements of the BALO-prey interaction, and it validates the hypothesis that mutational changes in a single gene can drastically impact prey resistance to BALO predation.

Background and Objectives: The microbial contamination of wastewater is associated with health risks. The aim of this study was to use the autochthonous Bdellovibrio potential to prey Gram-negative pathogenic bacteria as a bio-control agent to treat urban wastewater. Materials and Methods: Thirty-six raw sewage samples were collected for isolation of Bdellovibrio. Double layer plaque assay was used for isolation and the isolates were identified by microscopic examination and molecular analysis. To evaluate the predatory potential for decrease number of Gram-negative pathogenic bacteria, plaque perdition assay, reduction in host cells viability by colony-forming unit (CFU) counting, reduction in optical density (OD) in co-cultures and assay of killing efficiency were carried out. Also, the raw wastewater was treated by Bdellovibrio then the reduction in CFU counting and reduction in OD was evaluated. Results: Four strains of Bdellovibrio were isolated and were registered in Gene Bank. Clear plaques were observed after 3-6 days of incubation for all prey cells. The CFU enumerations of all preys were decreased after 48 hrs in co-cultures and raw wastewater. Also, OD was decreased down to 0.2 nm after 48 hrs. Conclusion: These autochthonous Bdellovibrio strains are proposed to use for bio-control of Gram-negative pathogenic bacteria in wastewater and reuse it for irrigation in arid regions.

The genus Acinetobacter has recently emerged as bacterial pathogens associated with mass mortalities in aquaculture. Yet scarce information is available on Acinetobacter venetianus as a potential pathogen of whiteleg shrimp Penaeus vannamei. In the present study, an A. venetianus pathogen (SHF1) was confirmed as a causative agent of red leg disease in freshwater-cultured whiteleg shrimp. It showed a mean lethal dose (LD50) value of 3.8 × 105 CFU mL−1 and developed multiple resistances to phenicols, quinolones and tetracycline drugs used in aquaculture. In addition, the pathogenic A. venetianus could be effectively controlled by Bdellovibrio powder at 0.8 mg L−1, which exhibited significant antibacterial effect and displayed good protection against A. venetianus challenge in P. vannamei for six days. As far as we know, this is the first study to identify A. venetianus as a potential bacterial pathogen of red leg disease in freshwater-farmed whiteleg shrimp.

The impact of key environmental factors, salinity, prey, and temperature, on the survival and ecology of Bdellovibrio and like bacteria (BALOs), including the freshwater/terrestrial, non-halotolerant group and the halophilic Halobacteriovorax strains, has been assessed based on a review of data in the literature. These topics have been studied by numerous investigators for nearly six decades now, and much valuable information has been amassed and reported. The collective data shows that salinity, prey, and temperature play a major role in, not only the growth and survival of BALOs, but also the structure and composition of BALO communities and the distribution of the predators. Salinity is a major determinant in the selection of BALO habitats, distribution, prey bacteria, and systematics. Halophilic BALOs require salt for cellular functions and are found only in saltwater habitats, and prey primarily on saltwater bacteria. To the contrary, freshwater/terrestrial BALOs are non-halotolerant and inhibited by salt concentrations greater than 0.5%, and are restricted to freshwater, soils, and other low salt environments. They prey preferentially on bacteria in the same habitats. The halophilic BALOs are further separated on the basis of their tolerance to various salt concentrations. Some strains are found in low salt environments and others in high salt regions. In situ studies have demonstrated that salinity gradients in estuarine systems govern the type of BALO communities that will persist within a specific gradient. Bacterial prey for BALOs functions more than just being a substrate for the predators and include the potential for different prey species to structure the BALO community at the phylotype level. The pattern of susceptibility or resistance of various bacteria species has been used almost universally to differentiate strains of new BALO isolates. However, the method suffers from a lack of uniformity among different laboratories. The use of molecular methods such as comparative analysis of the 16S rDNA gene and metagenomics have provided more specific approaches to distinguished between isolates. Differences in temperature growth range among different BALO groups and strains have been demonstrated in many laboratory experiments. The temperature optima and growth range for the saltwater BALOs is typically lower than that of the freshwater/terrestrial BALOs. The collective data shows not only that environmental factors have a great impact on BALO ecology, but also how the various factors affect BALO populations in nature.

We report the complete genome sequence of the predatory bacterium Bdellovibrio sp. strain KM01, isolated from soil collected near a pond. The genome is 3,961,288 bp long with 45.5% GC content. Comparative genomics among Bdellovibrio strains will help us understand how genotypic differences affect differences in predatory phenotypes.

No abstract available]

© 2020 Herencias et al., In this study we analyze the growth-phase dependent metabolic states of Bdellovibrio bacteriovorus by constructing a fully compartmented, mass and charge-balanced genome-scale metabolic model of this predatory bacterium (iCH457). Considering the differences between life cycle phases driving the growth of this predator, growth-phase condition-specific models have been generated allowing the systematic study of its metabolic capabilities. Using these computational tools, we have been able to analyze, from a system level, the dynamic metabolism of the predatory bacteria as the life cycle progresses. We provide computational evidences supporting potential axenic growth of B. bacteriovorus’s in a rich medium based on its encoded metabolic capabilities. Our systems-level analysis confirms the presence of “energy-saving” mechanisms in this predator as well as an abrupt metabolic shift between the attack and intraperiplasmic growth phases. Our results strongly suggest that predatory bacteria’s metabolic networks have low robustness, likely hampering their ability to tackle drastic environmental fluctuations, thus being confined to stable and predictable habitats. Overall, we present here a valuable computational testbed based on predatory bacteria activity for rational design of novel and controlled biocatalysts in biotechnological/clinical applications., This work was supported by the Spanish Ministry of Economy and Competitiveness through the grants BIO2014-59528-JIN (JN), BIO2013-44878-R, and BIO2017-83448-R and H2020 Engicoin no 760994 (MAP). CH was supported by a FPI (Contratos predoctorales para la formación de doctores) fellowship (BES-2014-070856). SSB was supported by a FPU (Ayuda para la formación de profesorado universitario) fellowship (FPU17/03978) from the Spanish Ministry of Universities.

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).

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.

Bacteria are preyed upon by diverse microbial predators, including bacteriophage and predatory bacteria, such as Bdellovibrio bacteriovorus While bacteriophage are used as antimicrobial therapies in Eastern Europe and are being applied for compassionate use in the United States, predatory bacteria are only just beginning to reveal their potential therapeutic uses. However, predation by either predator type can falter due to different adaptations arising in the prey bacteria. When testing poultry farm wastewater for novel Bdellovibrio isolates on Escherichia coli prey lawns, individual composite plaques were isolated containing both an RTP (rosette-tailed-phage)-like-phage and a B. bacteriovorus strain and showing central prey lysis and halos of extra lysis. Combining the purified phage with a lab strain of B. bacteriovorus HD100 recapitulated haloed plaques and increased killing of the E. coli prey in liquid culture, showing an effective side-by-side action of these predators compared to their actions alone. Using approximate Bayesian computation to select the best fitting from a variety of different mathematical models demonstrated that the experimental data could be explained only by assuming the existence of three prey phenotypes: (i) sensitive to both predators, (ii) genetically resistant to phage only, and (iii) plastic resistant to B. bacteriovorus only. Although each predator reduces prey availability for the other, high phage numbers did not abolish B. bacteriovorus predation, so both predators are competent to coexist and are causing different selective pressures on the bacterial surface while, in tandem, controlling prey bacterial numbers efficiently. This suggests that combinatorial predator therapy could overcome problems of phage resistance.IMPORTANCE With increasing levels of antibiotic resistance, the development of alternative antibacterial therapies is urgently needed. Two potential alternatives are bacteriophage and predatory bacteria. Bacteriophage therapy has been used, but prey/host specificity and the rapid acquisition of bacterial resistance to bacteriophage are practical considerations. Predatory bacteria are of interest due to their broad Gram-negative bacterial prey range and the lack of simple resistance mechanisms. Here, a bacteriophage and a strain of Bdellovibrio bacteriovorus, preyed side by side on a population of E. coli, causing a significantly greater decrease in prey numbers than either alone. Such combinatorial predator therapy may have greater potential than individual predators since prey surface changes selected for by each predator do not protect prey against the other predator.

A novel bacterial strain, designated YN101T, was isolated from a marine solar saltern in the coast of Weihai, Shandong Province, China. Strain YN101T was Gram-stain negative, facultatively anaerobic, oxidase and catalase negative bacterium with the ability to prey on other microbes. A cross-streaking culture method was utilized to analyze the predatory activity of strain YN101T. The results showed strain YN101T could prey on various bacteria, either Gram-stain negative or Gram-stain positive. According to the predatory assays, different species in the same genus may behave differently when attacked by strain YN101T. The predatory behavior of strain YN101T to four typical species was analyzed, and furthermore, predation to Algoriphagus marinus am2T were quantitatively studied by fluorogenic quantitative PCR, and the gene copies decreased over two magnitudes. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YN101T shared the greatest sequence similarity of 93.9% to Bradymonas sediminis FA350T. The complete genome sequence of strain YN101T was 8,047,306 bp in size and the genomic DNA G + C content was 63.8 mol%. The digital DNA-DNA hybridization (dDDH) values and average nucleotide identity (ANI) values between strain YN101T and B. sediminis FA350T were 13.9 and 74.0%. The genetic features showed that the biosynthesis of many important compounds was deficient in genome of strain YN101T, which may lead to its predation. Moreover, its genome encoded many genes affiliated with type IV pili, secretion system, membrane proteins and transduction proteins. Similar with myxobacteria and Bdellovibrio and like organisms (BALOs), these genes should play important roles in motility, adhesion or virulence to attack prey cells during predation. The predominant polar lipid profile of strain YN101T consisted of phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), and one unidentified aminophospholipid (APL). The major cellular fatty acid of strain YN101T was iso-C17:0, and the sole respiratory quinone was MK-7. Based on the chemotaxonomic, physiological and biochemical characteristics, strain YN101T represents a novel species of a novel genus in the family Bradymonadaceae, for which the name Persicimonas caeni gen. nov., sp. nov. is proposed. The type strain is YN101T (=KCTC 72083T = MCCC 1H00374T).

Bacterial predation, as exemplified by the periplasm-invading model predator Bdellovibrio bacteriovorus, is a fascinating multistage process facilitated by several adaptations to ‘regular’ bacterial lifestyles. We are beginning to understand more about such adaptations at the molecular level, particularly those concerning processes near the beginning of the predatory lifecycle (recognition, invasion, prey cell wall manipulation). In this review we highlight recent advances in investigating predation and tie these into some of the classical observations and phenotypes that make this two-cell system such an interesting case study in evolution.

To seek how soil biotic and abiotic factors which might shape the Bdellovibrio-and-like-organisms community, we sampled paddy soils under different fertilization treatments including fertilization without nitrogen (Control), the nitrogen use treatment (N) and the nitrogen overuse one (HNK) at three rice growing stages. The abundances of BALOs were impacted by the rice-growing stages but not the fertilization treatments. The abundances of Bdellovibrionaceae-like were positively associated with soil moisture, which showed a negative relationship with Bacteriovoracaceae-like bacteria. High-throughput sequencing analysis of the whole bacterial community revealed that the α-diversity of BALOs was not correlated with any soil properties data. Network analysis detected eight families directly linked to BALOs, namely, Pseudomonadaceae, Peptostreptococcaceae, Flavobacteriaceae, Sediment-4, Verrucomicrobiaceae, OM27, Solirubrobacteraceae and Roseiflexaceae. The richness and composition of OTUs in the eight families were correlated with different soil properties, while the evenness of them had a positive effect on the predicted BALO biomass. These results highlighted that the bottom-up control of BALOs in paddy soil at least partially relied on the changes of soil water content and the diversity of bacteria directly linked to BALOs in the microbial network.

Bdellovibrio-and-like organisms (BALOs) are a small group of bacteria that actively predate on other Gram-negative bacterial species. Although viewed mostly in a positive light, such as their potential use as living antibiotics to reduce pathogenic strain populations, several studies have also highlighted the need to control their activities, such as in the production of biodiesel. Consequently, this mini-review discusses research being conducted to characterize compounds and environmental settings that influence predation rates and the mechanisms by which they accomplish this, with a heavy emphasis on studies published within the last decade.Key points• This review discusses bacterial predators and factors impacting their activities. • Emphasis is on recent articles, particularly those discussing prey metabolites. • The implications on possible applications of bacterial predators are discussed.

In this study we analyze the growth-phase dependent metabolic states of Bdellovibrio bacteriovorus by constructing a fully compartmented, mass and charge-balanced genome-scale metabolic model of this predatory bacterium (iCH457). Considering the differences between life cycle phases driving the growth of this predator, growth-phase condition-specific models have been generated allowing the systematic study of its metabolic capabilities. Using these unprecedented computational tools, we have been able to analyze, from a system level, the dynamic metabolism of the predatory bacteria as the life cycle progresses. We provide solid computational evidences supporting potential axenic growth of B. bacteriovorus’s in a rich medium based its encoded metabolic capabilities. Our systems-level analysis confirms the presence of “energy-saving” mechanisms in this predator as well as an abrupt metabolic shift between the attack and intraperiplasmic growth phases. Our results strongly suggest that predatory bacteria’s metabolic networks have low robustness, likely hampering their ability to tackle drastic environmental fluctuations, thus being confined to stable and predictable habitats. Overall, we present here a valuable computational testbed based on predatory bacteria activity for rational design of novel and controlled biocatalysts in biotechnological/clinical applications.AUTHOR SUMMARYBacterial predation is an interspecific relationship widely extended in nature. Among other predators, Bdellovibrio and like organism (BALOs) have received recently a great attention due to the high potential applications such as biocontrol agent in medicine, agriculture, aquaculture and water treatment. Despite the increasing interest in of this predatory bacterium its complex lifestyle and growth conditions hamper the full exploitation of their biotechnological properties. In order to overcome these important shortcomings, we provide here the first genome-scale model of a predator bacterium constructed so far. By using the model as a computational testbed, we provide solid evidences of the metabolic autonomy of this interesting bacterium in term of growth, as well as its dynamic metabolism powering the biphasic life style of this predator. We found a low metabolic robustness thus suggesting that Bdellovibrio is more niche-specific than previously thought and that the environmental conditions governing predation may be relatively uniform. Overall, we provide here a valuable computational tool largely facilitating rational design of novel and controlled predator-based biocatalysts in biotechnological/clinical applications.

26 p.-4 fig.-2 tab. The Ecology of Predation at the Microscale pp 173-194, This chapter reviews the potential of the predatory bacteria Bdellovibrio bacteriovorus, an obligate predator of other gram-negative bacteria, as a biotechnological tool. Due to the unique lifestyle and the different applications, predatory bacteria have awakened interest to be developed as a lytic tool. The lack of physiological and metabolic information makes difficult this development. However, in the last years, different approaches have been described in order to understand the physiology, morphology, and metabolism of the predators, as well as the population dynamics of the prey-predator interactions. Besides its potential of “living antibiotic”, predatory bacteria have been proposed as a biocontrol agent in the food industry or aquaculture. A recent work using B. bacteriovorus as a biological lytic tool for the recovery of intracellular bioproducts highlighted the potential use of predators in industrial bioprocesses. The bottlenecks of using other Bdellovibrio and like organisms (BALOs) have been also considered and discussed during this chapter., This work was supported by the European Union’s Horizon 2020 Research and Innovation Programme, grant agreement no. 760994-2 (ENGICOIN), the Spanish Ministry of Science, Innovation and Universities (BIO2017-83448-R) and the Community of Madrid (P2018/ NMT4389).

‘Everything is everywhere, but the environment selects’ – Lourens Baas Becking This quote by Dutch botanist Lourens Baas Becking, when extended to microbiology, implies the activities of microorganisms are defined by the environment in which they find themselves, with biotic and abiotic factors alike impacting their activity, metabolism and viability. This is definitely true of Bdellovibrio bacteriovorus-and-like-organisms (BALOs), which are influenced directly by both their environment and the prey metabolic activities, stimuli that can have drastic impacts on the predatory activities of these strains. The goal of this chapter, therefore, is to delve deeper into, and gain a better understanding of, the correlation between predatory bacteria and their environment. Towards this end, we discuss here many of the factors and conditions (biotic and abiotic) that impact BALOs and their activities, including the osmolality, oxygen, serum albumin and indole. We hope through this discussion young scientists will gain a significant understanding of the current hurdles holding BALOs back from many real-world applications, such as in treatment of bacterial infections or within wastewater treatment systems, and encouragement to find solutions.

The Bdellovibrio and like organisms (BALOs) are obligate predators of gram negative bacteria, for which they have an absolute requirement in order to replicate and complete their life cycle. This peculiar life style profoundly affects their physiology and cellular biology and defines their ecology. BALOs are ubiquitous in soils and in water bodies; while they are not numerically dominant, they can be abundant and diverse. Among the water bodies, BALOs inhabit wastewater treatment plant reactors (WWTP) as well as other schemes where water is recycled. Their capacity to prey upon gram negative bacteria and their semi-generalist feeding suggest that they may play important roles in bacterial biomass turnover and in the reduction of numerous pathogens in these environments. This chapter first introduces the BALO’s life cycle, and features of their ecology and dynamics. It then presents our understanding of BALO community structure and dynamics under varying conditions in WWTPs, detailing latest research studies showing that BALO abundance is greatly influenced by the type of treatment applied to the wastewater. It ends by presenting open questions on our understanding of BALO effects on bacterial communities and by suggesting novels ways to address these questions, considering their capacity to improve wastewater treatment.

International audience; When considering microbial biotic interactions, viruses as well as eukaryotic grazers are known to be important components of aquatic microbial food webs. It might be the same for bacterivorous bacteria but these groups have been comparatively less studied. This is typically the case of the Bdellovibrio and like organisms (BALOs), which are obligate bacterial predators of other bacteria. Recently, the abundance and distribution of three families of this functional group were investigated in perialpine lakes, revealing their presence and quantitative importance. Here, a more in-depth analysis is provided for Lake Geneva regarding the diversity of these bacterial predators at different seasons, sites and depths. We reveal a seasonal and spatial (vertical) pattern for BALOs. They were also found to be relatively diverse (especially Bdellovibrionaceae) and assigned to both known and unknown phylogenetic clusters. At last we found that most BALOs were positively correlated to other bacterial groups, mainly Gram-negative, in particular Myxococcales (among which many are predators of other microbes). This study is the first shedding light on this potentially important bacterial killing group in a large and deep lake.

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 as candidates for the restoration of microbiomes and the prevention of dysbiosis.

In this paper, we firstly reviewed the diseases and antibiotic resistance problems facing the aquaculture industry, and introduced the probiotic concept and the use of Bdellovibrio and like organisms (BALOs) as a probiotic/biological agent to control pathogens or potential pathogens in a sustainable aquaculture. Then we reviewed, in greater details, the antibacterial activities of BALOs in aquaculture. As a special group of predator prokaryotic bacteria, BALOs are increasingly being regarded as an alternative biocontrol agent in the field of aquaculture, in China in particular. We have comprehensively evaluated relevant high quality documentations and discovered that BALOs are not only capable of lysing a wide range of Gram-negatives, but also some Gram-positives, depending on the strain/isolate specificities. Various challenge tests confirmed that BALOs do have protective effects on the organisms tested against the challenged pathogens. Furthermore, Pearson correlation coefficient analyses revealed that BALOs have significantly negative correlations with total bacterial and vibrio counts in rearing waters and the guts of reared organisms (p < 0.01), and positive correlations with their growth (body length and weight gains) and survival (p < 0.01). Finally, we concluded that BALOs do possess strong antibacterial activities against various bacteria, including vibrios, and have a very strong application potential in aquaculture practices.

Intensive studies have focused on external factors governing the gut microbiota-disease relationship. It remains unknown how internal gut microbial components, especially interkingdom predators — phagotrophs and Bdellovibrio and like organisms (BALOs) — are linked to host health. Herein, we integrated the holistic gut microbiome including pathogens, bacteria, protists, predators, functional pathways, and shrimp immune potentials, to identify the gut microbial components that were indicative of shrimp white feces syndrome (WFS) onset. Both diversities and structures of the gut BALOs and phagotrophs communities differed significantly at the WFS onset stage, but not of the gut bacterial and protistan counterparts. Pathogens dynamics were best predicted by the gut BALOs or phagotrophs. Notably, nine WFS discriminatory phagotrophic taxa contributed 78.6% diagnose accuracy of shrimp health status at the WFS onset stage. The differential functional pathways were intimately associated with BALOs. There were intensive interactions among pathogens, gut BALOs, phagotrophs, and shrimp immune potentials. Specifically, shrimp activate multiple signaling pathways in response to WFS. However, pathogens encode virulence factors that neutralize and evade shrimp immune clearance, thereby leading to WFS outbreak. Collectively, we proposed a conceptual model to infer the etiology of shrimp WFS. Our findings bridge the interplay among interkingdom predator-prey interactions, pathogens invasion, host immune feedbacks, and disease progression, which greatly broaden our understanding of shrimp WFS etiology from ecological and molecular perspectives.

Bdellovibriospp. are predatory bacteria with great potential as antimicrobial agents. Studies have shown that members of the genusBdellovibrioexhibit peculiar characteristics that influence their ecological adaptations. In this study, whole genomes of two differentBdellovibriospp. designated SKB1291214 and SSB218315 isolated from soil were sequenced. The core genes shared by all theBdellovibriospp. considered for the pangenome analysis including the epibioticB. exovoruswere 795. The number of unique genes identified inBdellovibriospp. SKB1291214, SSB218315, W, andB. exovorusJJS was 1343, 113, 857, and 1572, respectively. These unique genes encode hydrolytic, chemotaxis, and transporter proteins which might be useful for predation in theBdellovibriostrains. Furthermore, the twoBdellovibriostrains exhibited differences based on the % GC content, amino acid identity, and 16S rRNA gene sequence. The 16S rRNA gene sequence ofBdellovibriosp. SKB1291214 shared 99% identity with that of an unculturedBdellovibriosp. clone 12L 106 (a pairwise distance of 0.008) and 95–97% identity (a pairwise distance of 0.043) with that of other culturable terrestrialBdellovibriospp., including strain SSB218315. InBdellovibriosp. SKB1291214, 174 bp sequence was inserted at the host interaction(hit)locus region usually attributed to prey attachment, invasion, and development of host independentBdellovibriophenotypes. Also, a gene equivalent toBd0108inB. bacteriovorusHD100 was not conserved inBdellovibriosp. SKB1291214. The results of this study provided information on the genetic characteristics and diversity of the genusBdellovibriothat can contribute to their successful applications as a biocontrol agent.

An investigation on the soil microbe populations from an agricultural inning area at the Pearl River estuary in Guangdong province, China was conducted via high through-put sequencing. The results revealed abundant diversity in the soil bacterial and fungal populations. In total, 197103 sequence tags were obtained from soil samples, most of which represented bacterial genera Actinomycetes, Bacillus and Marinobacter, while a majority of 118378 tags obtained were derived from fungal genera Clostridium, Devosia, Bradyrhizobium, Bdellovibrio, Phenylobacterium, Penicillium, and Emericella. Furthermore, nine physiological indexes (pH, available phosphorous, basic-group nitrogen, available potassium, catalase, sucrose, urease, phosphatase, and organic matters) were measured in three soil samples, and the association between these physiological indexes and microbe population composition was examined. The results revealed obvious inter-sample differences associated with ten dominating microbial groups: genera Clostridium, Devosia, Bradyrhizobium, Bdellovibrio, Phenylobacterium, and Penicillium were mainly impacted by pH (with a positive correlation), genera Sphingomonas and Acinetobacter mainly by available phosphorous (positive correlation), and genera Gemmatimonas and Pseudomonas by both pH and available phosphorous (negative correlation). Our study suggested that regulation of microbial species/populations might help improve soil environment to facilitate the growth of crops’ above-ground parts, and this provides practical information for inning agriculture.

For comprehensive insights into the effects of disinfection strategy on antibiotic resistance genes (ARGs) in pipe wall biofilm to ensure the biosafety of domestic hot water system (DHWS), this study investigated the occurrence, removal and interactions of ARGs, as well as their potential hosts. The shifts in biofilm community composition and microbial functions (level three) were observed under different disinfection strategies. A total of 40 ARG subtypes within seven antibiotic types were predicted. The total gene counts ranged from 6.10 × 104 to 2.31 × 105, and decreased after the chlorine, chlorine dioxide, silver ions (Ag+), ultraviolet (UV), and Ag+-UV disinfection. The ARGs in samples under different disinfection strategies were divided into four groups, and the removal and enrichment of the same ARGs in each group were almost consistent. The complex interactions among ARGs were weakened and altered by disinfection (edges decreased from 237 to 178), with yebQ and marC as the hub ARGs, but positive correlations were still dominant. Moreover, 36 nonpathogenic bacteria, including high- (Blastocatella), mid- (Ornatilinea) and low-abundant genera (Bdellovibrio), as well as 18 potential pathogens (Vibrio and Bacillus) were identified as the potential hosts harboring ARGs. Overall, this study provides some new insights into the occurrence of ARGs and their fate under eight disinfection strategies, which is of great significance for a comprehensive understanding and control of ARGs in DHWS.

Modification of essential bacterial peptidoglycan (PG) containing cell walls can lead to antibiotic resistance, for example β-lactam resistance by L,D-transpeptidase activities. Predatory Bdellovibrio bacteriovorus are naturally antibacterial and combat infections by traversing, modifying and finally destroying walls of Gram-negative prey bacteria, modifying their own PG as they grow inside prey. Historically, these multi-enzymatic processes on two similar PG walls have proved challenging to elucidate. Here, with a PG labelling approach utilizing timed pulses of multiple fluorescent D-amino acids (FDAAs), we illuminate dynamic changes that predator and prey walls go through during the different phases of bacteria:bacteria invasion. We show formation of a reinforced circular port-hole in the prey wall; L,D-transpeptidaseBd mediated D-amino acid modifications strengthening prey PG during Bdellovibrio invasion and a zonal mode of predator-elongation. This process is followed by unconventional, multi-point and synchronous septation of the intracellular Bdellovibrio, accommodating odd- and even-numbered progeny formation by non-binary division.

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.

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.

Bdellovibrio bacteriovorus is an obligate predator of bacteria that grows and divides within the periplasm of its prey. Functions involved in the early steps of predation have been identified and characterized, but mediators of prey invasion are still poorly detailed. By combining omics data available for Bdellovibrio and like organisms (BALO’s), we identified 43 genes expressed in B. bacteriovorus during the early interaction with prey. These included genes in a tight adherence (TAD) operon encoding for two type IVb fimbriae-like pilin proteins (flp1 and flp2), and their processing and export machinery. Two additional flp genes (flp3 and flp4) were computationally identified at other locations along the chromosome, defining the largest and most diverse type IVb complement known in bacteria to date. Only flp1, flp2 and flp4 were expressed; their respective gene knock-outs resulted in a complete loss of the predatory ability without losing the ability to adhere to prey cells. Additionally, we further demonstrate differential regulation of the flp genes as the TAD operon of BALOs with different predatory strategies is controlled by a flagellar sigma factor FliA, while flp4 is not. Finally, we show that FliA, a known flagellar transcriptional regulator in other bacteria, is an essential Bdellovibrio gene.

In this study, Bdellovibrio and like organisms (BALOs) strain BDH12 that possessed a wide lysis spectrum was tested in coldwater abalone (Haliotis discus hannai Ino) juvenile rearing, and its effect on survival and growth and on gut bacterial communities was investigated. A 90-day rearing experiment at the production level was performed. At the end, juvenile abalone grew from the initial shell length of 4.0 ± 1.0 to 12.63 ± 2.12 mm in control and 14.89 ± 1.27 mm in test (p