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Bdellovibrio bacteriovorus, an obligate predator of Gram-negative bacteria, has emerged as a promising natural antibiotic to combat the escalating threat of antibiotic resistance. Plaque forming units (PFU) counting is commonly used to determine the viable numbers of B. bacteriovorus. However, nearly three-days incubation is always necessary for getting the single, obvious plaques on the double-layer agar plate. This time-consuming procedure greatly impedes the purification and enumeration efficiency of B. bacteriovorus. Until now, no simplified method has been developed to address this issue. In this work, we evaluated the advantages of using fluorescent prey to enhance the visualization of the B. bacteriovorus plaques. Our work reveals that the regular single plaques are easily observed under the enhanced background of fluorescent prey lawn in one and a half days, reducing nearly half of the time consumption in the purification and enumeration of B. bacteriovorus. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance (AMR) is an increasing problem worldwide, with significant associated morbidity and mortality. Given the slow production of new antimicrobials, non-antimicrobial methods for treating infections with significant AMR are required. This review examines the potential of predatory bacteria to combat infectious diseases, particularly those caused by pathogens with AMR. Predatory bacteria already have well-known applications beyond medicine, such as in the food industry, biocontrol, and wastewater treatment. Regarding their potential for use in treating infections, several in vitro studies have shown their potential in eliminating various pathogens, including those resistant to multiple antibiotics, and they also suggest minimal immune stimulation and cytotoxicity by predatory bacteria. In vivo animal studies have demonstrated safety and efficacy in reducing bacterial burden in various infection models. However, results can be inconsistent, suggesting dependence on factors like the animal model and the infecting bacteria. Until now, no clinical study in humans exists, but as experience with predatory bacteria grows, future studies including clinical studies in humans could be designed to evaluate their efficacy and safety in humans, thus leading to the potential for approval of a novel method for treating infectious diseases by bacteria.

Antimicrobial resistance (AMR) is an increasing problem worldwide, with significant associated morbidity and mortality. Given the slow production of new antimicrobials, non-antimicrobial methods for treating infections with significant AMR are required. This review examines the potential of predatory bacteria to combat infectious diseases, particularly those caused by pathogens with AMR. Predatory bacteria already have well-known applications beyond medicine, such as in the food industry, biocontrol, and wastewater treatment. Regarding their potential for use in treating infections, several in vitro studies have shown their potential in eliminating various pathogens, including those resistant to multiple antibiotics, and they also suggest minimal immune stimulation and cytotoxicity by predatory bacteria. In vivo animal studies have demonstrated safety and efficacy in reducing bacterial burden in various infection models. However, results can be inconsistent, suggesting dependence on factors like the animal model and the infecting bacteria. Until now, no clinical study in humans exists, but as experience with predatory bacteria grows, future studies including clinical studies in humans could be designed to evaluate their efficacy and safety in humans, thus leading to the potential for approval of a novel method for treating infectious diseases by bacteria. [ABSTRACT FROM AUTHOR]

Identifying the nutritional requirements and growth conditions of microorganisms is crucial for determining their applicability in industry and understanding their role in clinical ecology. Predatory bacteria such as Bdellovibrio bacteriovorus have emerged as promising tools for combating infections by human bacterial pathogens due to their natural killing features. Bdellovibrio's lifecycle occurs inside prey cells, using the cytoplasm as a source of nutrients and energy. However, this lifecycle supposes a challenge when determining the specific uptake of metabolites from the prey to complete the growth inside cells, a process that has not been completely elucidated. Here, following a model-based approach, we illuminate the ability of B. bacteriovorus to replicate DNA, increase biomass, and generate adenosine triphosphate (ATP) in an amino acid-based rich media in the absence of prey, keeping intact its predatory capacity. In this culture, we determined the main carbon sources used and their preference, being glutamate, serine, aspartate, isoleucine, and threonine. This study offers new insights into the role of predatory bacteria in natural environments and establishes the basis for developing new Bdellovibrio applications using appropriate metabolic and physiological methodologies. Key points: • Amino acids support axenic lifestyle of Bdellovibrio bacteriovorus. • B. bacteriovorus preserves its predatory ability when growing in the absence of prey. [ABSTRACT FROM AUTHOR]

IntroductionThe intricate habitats of aquatic organisms, coupled with the prevalence of pathogens, contribute to a high incidence of various diseases, particularly bacterial infections. Consequently, the formulation of sustainable and effective disease management strategies is crucial for the thriving aquaculture sector.Methods and resultsIn this investigation, a strain of Bdellovibrio bacteriovorus, designated B. bacteriovorus FWA, was isolated from a freshwater fish pond. Identification was achieved through microscopic examination of morphological characteristics, biochemical property assessment, and phylogenetic analysis. The lysogenic capability of B. bacteriovorus FWA was evaluated, revealing its effectiveness in lysing Escherichia coli, Aeromonas hydrophila, Vibrio alginolyticus, Vibrio parahaemolyticus, and Edwardsiella tarda. Physiological analysis indicated that the optimal ratio of B. bacteriovorus FWA to host bacteria was 1:10,000, with strict aerobic requirements. The optimal pH range for growth and reproduction was 7.0–8.0, the ideal temperature was found to be 30–35°C, with a preferred Na+ concentration of 0% and a Ca2+ concentration of 15–25 mM. Additionally, B. bacteriovorus FWA demonstrated enhanced lytic activity against bacteria in aquaculture effluent while effectively managing ammonia-nitrogen levels.DiscussionIn summary, B. bacteriovorus FWA holds significant promise for development as a probiotic agent in aquaculture.

The development of potent bactericidal antibiotic alternatives is important to address the current antibiotic crisis. A representative example is the topical delivery of predatory Bdellovibrio bacteriovorus bacteria to treat ocular bacterial infection. However, the direct topical use of B. bacteriovorus suspensions has the problem of easy loss and inactivation. Here, a B. bacteriovorus in situ forming hydrogel (BIG) was constructed for the ocular delivery of B. bacteriovorus. BIGs, as a fluid in their primitive state, were temperature- and cation- dually sensitive, which was rapidly transformed into immobile gels in the ocular environment. BIGs not only kept the activity of B. bacteriovorus but also retained on the ocular surface for a long time. The biosafety of BIGs was good without HCEC cell toxicity and hemolysis. More importantly, BIGs highly inhibited the growth of drug-resistant Pseudomonas aeruginosa whether in vitro or in the infected rat eyes. The ocular infection was completely controlled by BIGs with no corneal ulcers and inflammations. This living bacteria gel is a promising medication for the local treatment of drug-resistant bacteria-induced ocular infection. [Display omitted] • Bdellovibrio bacteriovorus in situ forming hydrogels (BIGs) formed from poloxamer 407/deacetylated gellan gum • Biocompatibility, high in vitro antibacterial ability, and long-term corneal retention of BIGs • High in vivo antibacterial and infected-corneal healing ability of BIGs [ABSTRACT FROM AUTHOR]

Bdellovibrio bacteriovorus is a bacterial agent that stands out for its ability to act as a predator against gram-negative bacteria and has found application against antibiotic-resistant pathogens. The aim of this study is to determine the efficacy of Bdellovibrio bacteriovorus against antibiotic-resistant pathogens, particularly those causing infections in surgical incision sites. A total of 6 experimental groups were created in mice, and surgical area infections were initiated with Klebsiella pneumoniae in incision sites. The effects of antibiotics and Bdellovibrio bacteriovorus alone or in combination were compared to the control group. In the Bdellovibrio bacteriovorus treatment group, edema and redness were observed in all mice at 24th hours, in 20% of mice at 48th hours, and in none at the 72 nd h. A significant difference was observed in the Bdellovibrio bacteriovorus treatment groups in reducing Klebsiella pneumoniae burden in the incision area compared to antibiotics alone or Bdellovibrio bacteriovorus + antibiotics, (p < 0.001). Likewise, cytokine level determinations indicated that B. bacteriovorus applications generated a therapeutic response without inducing an inflammatory response. • Surgical Site Infections (SSI) are the most common healthcare-associated infections, leading to increased workload and cost, as they prolong hospital stays, increase in morbidity and mortality, cause readmissions, and may require reoperation. • Infections caused by antibiotic-resistant gram-negative bacteria in SSI have been increasing in recent years and are reported to be associated with high levels of morbidity and mortality. • Resistance to antibiotic therapy used against gram-negative bacteria causes treatment to become increasingly difficult and increases the need for new types of treatments in clinics. • One of these approaches, which is the use of live bacteria such as Bdellovibrio bacteriovorus (B. bacteriovorus) to treat antibiotic-resistant pathogens, is considered one of the most promising possibilities. • This is the first study evaluating the efficacy of B. bacteriovorus on surgical site infections in experimental animals. [ABSTRACT FROM AUTHOR]

Wound infection is becoming a considerable healthcare crisis due to the abuse of antibiotics and the substantial production of multidrug-resistant bacteria. Seawater immersion wounds usually become a mortal trouble because of the infection of Vibrio vulnificus. Bdellovibrio bacteriovorus, one kind of natural predatory bacteria, is recognized as a promising biological therapy against intractable bacteria. Here, we prepared a B. bacteriovorus-loaded polyvinyl alcohol/alginate hydrogel for the topical treatment of the seawater immersion wounds infected by V. vulnificus. The B. bacteriovorus-loaded hydrogel (BG) owned highly microporous structures with the mean pore size of 90 μm, improving the rapid release of B. bacteriovorus from BG when contacting the aqueous surroundings. BG showed high biosafety with no L929 cell toxicity or hemolysis. More importantly, BG exhibited excellent in vitro anti-V. vulnificus effect. The highly effective infected wound treatment effect of BG was evaluated on mouse models, revealing significant reduction of local V. vulnificus, accelerated wound contraction, and alleviated inflammation. Besides the high bacterial inhibition of BG, BG remarkably reduced inflammatory response, promoted collagen deposition, neovascularization and re-epithelization, contributing to wound healing. BG is a promising topical biological formulation against infected wounds.

Bdellovibrio bacteriovorus is a predatory gram-negative delta-proteobacteria that preys on other gram-negative bacterial species. With the surge in the cases of Anti-Microbial Resistance and the demand to replace conventional antibiotics, this predatory attribute of B. bacteriovorus has led to a vast number of studies in the last decade alone. These studies demonstrated various aspects of predation and proposed using B. bacteriovorus as an alternative new approach to conventional antibiotics in humans. Here we review the current progress made in this promising new area of research to develop novel antibacterial approaches.

Since its discovery six decades ago, the predatory bacterium Bdellovibrio bacteriovorus has sparked recent interest as a potential remedy to the antibiotic resistance crisis. Here we give a comprehensive historical overview from discovery to progressive developments in microscopy and molecular mechanisms. Research on B. bacteriovorus has moved from curiosity to a new model organism, revealing over time more details on its physiology and fascinating predatory life cycle with the help of a variety of methods. Based on recent findings in cryo-electron tomography, we recapitulate on the intricate molecular details known in the predatory life cycle including how this predator searches for its prey bacterium, to how it attaches, grows, and divides all from within the prey cell. Finally, the newly developed B. bacteriovorus progeny leave the prey cell remnants in the exit phase. While we end with some unanswered questions remaining in the field, new imaging technologies and quantitative, systematic advances will likely help to unravel them in the next decades. [ABSTRACT FROM AUTHOR]

We tested the predation of B. bacteriovorus against Escherichia coli in milk samples in three different experiments. In Experiment 1, the growth and predatory activity of B. bacteriovorus against E. coli in milk stored at 4°C were evaluated. In Experiment 2, the predatory activity of B. bacteriovorus against E. coli in the milk matrix was compared to the optimal one in the medium of choice. In Experiment 3, the influence of the native milk microbial community on the predation of B. bacteriovorus against E. coli experimentally added or indigenous grown at 4°C was tested. The predator increased at 4°C by about 1 Log in the first 48 hours and caused E. coli decrease by about 2 Log after 24 hours. The predator at 30°C reduced E. coli faster (3 Log after 6 hours) than at 4°C (2 Log after 24 hours). B. bacteriovorus at 30°C preyed on E. coli more in the nutrient broth than in the milk, with the most significant difference by about 4 Log after 48 hours. In raw milk contaminated only by the predator, it increased by about 1 Log after 48 hours at 4°C, suggesting that it preyed on indigenous microorganisms. B. bacteriovorus could find application in raw milk used as food or raw material during storage at 4°C to reduce the microbial load of spoilage and Shigatoxin-producing E. coli (STEC) strains of E. coli and therefore increase its shelf-life and healthiness. [ABSTRACT FROM AUTHOR]

ABSTRACT Most bacteria, including model organisms such as Escherichia coli, Bacillus subtilis, and Caulobacter crescentus, reproduce by binary fission. However, some bacteria belonging to various lineages, including antibiotic-producing Streptomyces and predatory Bdellovibrio, proliferate by nonbinary fission, wherein three or more chromosome copies are synthesized and the resulting multinucleoid filamentous cell subdivides into progeny cells. Here, we demonstrate for the first time that the predatory bacterium Bdellovibrio bacteriovorus reproduces through both binary and nonbinary fission inside different prey bacteria. Switching between the two modes correlates with the prey size. In relatively small prey cells, B. bacteriovorus undergoes binary fission; the FtsZ ring assembles in the midcell, and the mother cell splits into two daughter cells. In larger prey cells, B. bacteriovorus switches to nonbinary fission and creates multiple asynchronously assembled FtsZ rings to produce three or more daughter cells. Completion of bacterial cell cycle critically depends on precise spatiotemporal coordination of chromosome replication with other cell cycle events, including cell division. We show that B. bacteriovorus always initiates chromosome replication at the invasive pole of the cell, but the spatiotemporal choreography of subsequent steps depends on the fission mode and/or the number of progeny cells. In nonbinary dividing filaments producing five or more progeny cells, the last round(s) of replication may also be initiated at the noninvasive pole. Altogether, we find that B. bacteriovorus reproduces through bimodal fission and that extracellular factors, such as the prey size, can shape replication choreography, providing new insights about bacterial life cycles. IMPORTANCE Most eukaryotic and bacterial cells divide by binary fission, where one mother cell produces two progeny cells, or, rarely, by nonbinary fission. All bacteria studied to date use only one of these two reproduction modes. We demonstrate for the first time that a predatory bacterium, Bdellovibrio bacteriovorus, exhibits bimodal fission and the mode of division depends on the size of the prey bacterium inside which B. bacteriovorus grows. This work provides key insights into the mode and dynamics of B. bacteriovorus proliferation in different pathogens that pose a major threat to human health due to their emerging antibiotic resistance (Proteus mirabilis, Salmonella enterica, and Shigella flexneri). The use of predatory bacteria such as B. bacteriovorus is currently regarded as a promising strategy to kill antibiotic-resistant pathogens. We find that B. bacteriovorus employs different chromosome replication choreographies and division modes when preying on those pathogens. Our findings may facilitate the design of efficient pathogen elimination strategies.

Since its discovery six decades ago, the predatory bacterium Bdellovibrio bacteriovorus has sparked recent interest as a potential remedy to the antibiotic resistance crisis. Here we give a comprehensive historical overview from discovery to progressive developments in microscopy and molecular mechanisms. Research on B. bacteriovorus has moved from curiosity to a new model organism, revealing over time more details on its physiology and fascinating predatory life cycle with the help of a variety of methods. Based on recent findings in cryo-electron tomography, we recapitulate on the intricate molecular details known in the predatory life cycle including how this predator searches for its prey bacterium, to how it attaches, grows, and divides all from within the prey cell. Finally, the newly developed B. bacteriovorus progeny leave the prey cell remnants in the exit phase. While we end with some unanswered questions remaining in the field, new imaging technologies and quantitative, systematic advances will likely help to unravel them in the next decades.

Antimicrobial resistance has become one of the most important public health problems of our century. In addition to the spread of resistance, biofilm production also makes the treatment of infections increasingly difficult. Therefore, this study, it was aimed to investigate the effect of the predator bacterium Bdellovibrio bacteriovorus HD100 on various clinical pathogens and their biofilms. A large panel of Gram-positive and negative clinical isolates were included in the study. The double-layer agar method was used to optimize the cultivation of predatory bacteria. The effectiveness of Bdellovibrio bacteriovorus HD 100 on planktonic cells and biofilms, was determined by co-culture and crystal violet staining methods, respectively. The antibiofilm activity was also visualized via scanning electron microscopy. The predator bacteria was found effective against most of the Gram-negative isolates. But it was determined that the lowest activity among these isolates was shown to Pseudomonas aeruginosa and Acinetobacter baumannii. Although it is known that B. bacteriovorus does not predate on Gram-positive isolates, interestingly, Staphylococci species included in this study were found to be inhibited in co-culture studies. As determined in co-culture and biofilm studies, B. bacteriovorus can be used to control both bacterial growth and biofilms in most Gram-negative species. Interestingly, our data also suggest that predatory bacteria may also be effective against Gram-positive bacterial biofilms in addition to Staphylococcus aureus. Although the evaluation of different species of isolates in this study demonstrates the potential of predatory bacteria, the host specificity and the relation of prey and predator need to be demonstrated. [ABSTRACT FROM AUTHOR]

Wound infection is becoming a considerable healthcare crisis due to the abuse of antibiotics and the substantial production of multidrug-resistant bacteria. Seawater immersion wounds usually become a mortal trouble because of the infection of Vibrio vulnificus. Bdellovibrio bacteriovorus , one kind of natural predatory bacteria, is recognized as a promising biological therapy against intractable bacteria. Here, we prepared a B. bacteriovorus -loaded polyvinyl alcohol/alginate hydrogel for the topical treatment of the seawater immersion wounds infected by V. vulnificus. The B. bacteriovorus- loaded hydrogel (BG) owned highly microporous structures with the mean pore size of 90 μm, improving the rapid release of B. bacteriovorus from BG when contacting the aqueous surroundings. BG showed high biosafety with no L929 cell toxicity or hemolysis. More importantly, BG exhibited excellent in vitro anti- V. vulnificus effect. The highly effective infected wound treatment effect of BG was evaluated on mouse models, revealing significant reduction of local V. vulnificus , accelerated wound contraction, and alleviated inflammation. Besides the high bacterial inhibition of BG, BG remarkably reduced inflammatory response, promoted collagen deposition, neovascularization and re-epithelization, contributing to wound healing. BG is a promising topical biological formulation against infected wounds. Dual-network Bdellovibrio bacteriovorus- loaded hydrogels (BG) were prepared for highly effective loading B. bacteriovorus and the treatment of bacterium-infected wounds by lysis of bacteria, reduction of inflammatory response, and improvement of collagen deposition, neovascularization and re-epithelization. [Display omitted] [ABSTRACT FROM AUTHOR]

With increasing levels of antimicrobial resistance impacting both human and animal health, novel means of treating resistant infections are urgently needed. Bacteriophages and predatory bacteria such as Bdellovibrio bacteriovorus have been proposed as suitable candidates for this role. Microbes also play a key environmental role as producers or recyclers of nutrients such as carbon and nitrogen, and predators have the capacity to be keystone species within microbial communities. To date, many studies have looked at the mechanisms of action of prokaryotic predators, their safety in in vivo models and their role and effectiveness under specific conditions. Mathematical models however allow researchers to investigate a wider range of scenarios, including aspects of predation that would be difficult, expensive, or time-consuming to investigate experimentally. We review here a history of modelling in prokaryote predation, from simple Lotka-Volterra models, through increasing levels of complexity, including multiple prey and predator species, and environmental and spatial factors. We consider how models have helped address questions around the mechanisms of action of predators and have allowed researchers to make predictions of the dynamics of predator-prey systems. We examine what models can tell us about qualitative and quantitative commonalities or differences between bacterial predators and bacteriophage or protists. We also highlight how models can address real-world situations such as the likely effectiveness of predators in removing prey species and their potential effects in shaping ecosystems. Finally, we look at research questions that are still to be addressed where models could be of benefit. [ABSTRACT FROM AUTHOR]

Bdellovibrio bacteriovorus is a predatory gram-negative delta-proteobacteria that preys on other gram-negative bacterial species. With the surge in the cases of Anti-Microbial Resistance and the demand to replace conventional antibiotics, this predatory attribute of B. bacteriovorus has led to a vast number of studies in the last decade alone. These studies demonstrated various aspects of predation and proposed using B. bacteriovorus as an alternative new approach to conventional antibiotics in humans. Here we review the current progress made in this promising new area of research to develop novel antibacterial approaches. [ABSTRACT FROM AUTHOR]

Bdellovibrio bacteriovorus, an obligate predator of Gram-negative bacteria, has emerged as a promising natural antibiotic to combat the escalating threat of antibiotic resistance. Plaque forming units (PFU) counting is commonly used to determine the viable numbers of B. bacteriovorus. However, nearly three-days incubation is always necessary for getting the single, obvious plaques on the double-layer agar plate. This time-consuming procedure greatly impedes the purification and enumeration efficiency of B. bacteriovorus. Until now, no simplified method has been developed to address this issue. In this work, we evaluated the advantages of using fluorescent prey to enhance the visualization of the B. bacteriovorus plaques. Our work reveals that the regular single plaques are easily observed under the enhanced background of fluorescent prey lawn in one and a half days, reducing nearly half of the time consumption in the purification and enumeration of B. bacteriovorus., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Drug-resistant pathogens significantly threaten human health and life. Simply killing drug-resistant pathogens cannot effectively eliminate their threat since the drug-resistant genes (DRGs) released from dead drug-resistant pathogens are difficult to eliminate and can further spread via horizontal gene transfer, leading to the spread of drug resistance. The development of antibacterial materials with sterilization and DRGs cleavage activities is highly crucial. Herein, a living system, Ce-PEA@Bdello, is fabricated with bacterial killing and DRGs cleavage activities for blocking bacterial drug resistance dissemination by engineered Bdellovibrio bacteriovorus (Bdello). Ce-PEA@Bdello is obtained by engineering Bdello with dopamine and a multinuclear cerium (IV) complex. Ce-PEA@Bdello can penetrate and eliminate kanamycin-resistant P. aeruginosa (Kan R ) biofilms via the synergistic effect of predatory Bdello and photothermal polydopamine under near-infrared light. Additionally, the DNase-mimicking ability of Ce-PEA@Bdello endows it with genome and plasmid DNA cleavage ability. An in vivo study reveals that Ce-PEA@Bdello can eliminate P. aeruginosa (Kan R ) and cleave DRGs in scald/burn infected wounds to block the spread of drug resistance and accelerate wound healing. This bioactive system constructed from natural living materials offers a promising means for blocking the spread of drug resistance., (© 2024 Wiley‐VCH GmbH.)

With increasing levels of antimicrobial resistance impacting both human and animal health, novel means of treating resistant infections are urgently needed. Bacteriophages and predatory bacteria such as Bdellovibrio bacteriovorus have been proposed as suitable candidates for this role. Microbes also play a key environmental role as producers or recyclers of nutrients such as carbon and nitrogen, and predators have the capacity to be keystone species within microbial communities. To date, many studies have looked at the mechanisms of action of prokaryotic predators, their safety in in vivo models and their role and effectiveness under specific conditions. Mathematical models however allow researchers to investigate a wider range of scenarios, including aspects of predation that would be difficult, expensive, or time-consuming to investigate experimentally. We review here a history of modelling in prokaryote predation, from simple Lotka-Volterra models, through increasing levels of complexity, including multiple prey and predator species, and environmental and spatial factors. We consider how models have helped address questions around the mechanisms of action of predators and have allowed researchers to make predictions of the dynamics of predator–prey systems. We examine what models can tell us about qualitative and quantitative commonalities or differences between bacterial predators and bacteriophage or protists. We also highlight how models can address real-world situations such as the likely effectiveness of predators in removing prey species and their potential effects in shaping ecosystems. Finally, we look at research questions that are still to be addressed where models could be of benefit.

Bdellovibrio, δ-proteobacteria, including B. bacteriovorus (Bba) and B. exovorus (Bex), are obligate predators of other Gram-negative bacteria. While Bba grows in the periplasm of the prey cell, Bex grows externally. We have analyzed and compared the transport proteins of these 2 organisms based on the current contents of the Transporter Classification Database (TCDB; www.tcdb.org). Bba has 103 transporters more than Bex, 50% more secondary carriers, and 3 times as many MFS carriers. Bba has far more metabolite transporters than Bex as expected from its larger genome, but there are 2 times more carbohydrate uptake and drug efflux systems, and 3 times more lipid transporters. Bba also has polyamine and carboxylate transporters lacking in Bex. Bba has more than twice as many members of the Mot-Exb family of energizers, but both may have energizers for gliding motility. They use entirely different types of systems for iron acquisition. Both contain unexpectedly large numbers of pseudogenes and incomplete systems, suggesting that they are undergoing genome size reduction. Interestingly, all 5 outer-membrane receptors in Bba are lacking in Bex. The 2 organisms have similar numbers and types of peptide and amino acid uptake systems as well as protein and carbohydrate secretion systems. The differences observed correlate with and may account, in part, for the different lifestyles of these 2 bacterial predators.

Bdellovibrio, δ-proteobacteria, including B. bacteriovorus (Bba) and B. exovorus (Bex), are obligate predators of other Gram-negative bacteria. While Bba grows in the periplasm of the prey cell, Bex grows externally. We have analyzed and compared the transport proteins of these 2 organisms based on the current contents of the Transporter Classification Database (TCDB; www.tcdb.org). Bba has 103 transporters more than Bex, 50% more secondary carriers, and 3 times as many MFS carriers. Bba has far more metabolite transporters than Bex as expected from its larger genome, but there are 2 times more carbohydrate uptake and drug efflux systems, and 3 times more lipid transporters. Bba also has polyamine and carboxylate transporters lacking in Bex. Bba has more than twice as many members of the Mot-Exb family of energizers, but both may have energizers for gliding motility. They use entirely different types of systems for iron acquisition. Both contain unexpectedly large numbers of pseudogenes and incomplete systems, suggesting that they are undergoing genome size reduction. Interestingly, all 5 outer-membrane receptors in Bba are lacking in Bex. The 2 organisms have similar numbers and types of peptide and amino acid uptake systems as well as protein and carbohydrate secretion systems. The differences observed correlate with and may account, in part, for the different lifestyles of these 2 bacterial predators.

Among predatory bacteria, Bdellovibrio and Like Organisms (BALOs) have a high potential for application as biocontrol agents. The objective of this study was to isolate and characterize native BALO strains from different water and soil samples in Iran. Nineteen BALO strains were obtained from the samples using Escherichia coli as prey. Analysis of 16S rRNA gene sequences showed that Na2, Tal2, Mm1, and Ja2 strains belong to a different species than B. bacteriovorus HD100 as a type strain. Also, Z2, B2, Ch2, Kan1, and Pj strains could be separate from the Bdellovibrio genera. The strains showed different host range and predation efficiency when they were tested with 17 different Gram-negative bacteria subsequently. The prey range of six strains including Sar, Pj, Kan1, C1, Z2, and Ch2 was limited to only E. coli. The most versatile predators were strains Zar3 and Kash2 that exhibited almost the same prey range as type strain HD100. They are also placed close to each other in the phylogenetic tree. The host range of strain Na2 was narrow and attacked only Ralstonia solanacearum, which is less susceptible to predation by the other strains. Native BALOs from water and soils in Iran were isolated, forming a group of potential biocontrol agents adapted to local conditions. [ABSTRACT FROM AUTHOR]

The survival, proliferation, and epidemic spread of Acinetobacter baumannii (A. baumannii) in hospital settings is associated with several characteristics, including resistance to many commercially available antibiotics as well as the expression of multiple virulence mechanisms. This severely limits therapeutic options, with increased mortality and morbidity rates recorded worldwide. The World Health Organisation, thus, recognises A. baumannii as one of the critical pathogens that need to be prioritised for the development of new antibiotics or treatment. The current review will thus provide a brief overview of the antibiotic resistance and virulence mechanisms associated with A. baumannii's "persist and resist strategy". Thereafter, the potential of biological control agents including secondary metabolites such as biosurfactants [lipopeptides (surfactin and serrawettin) and glycolipids (rhamnolipid)] as well as predatory bacteria (Bdellovibrio bacteriovorus) and bacteriophages to directly target A. baumannii, will be discussed in terms of their in vitro and in vivo activity. In addition, limitations and corresponding mitigations strategies will be outlined, including curtailing resistance development using combination therapies, product stabilisation, and large-scale (up-scaling) production. [ABSTRACT FROM AUTHOR]

The interaction of Bdellovibrio bacteriovorus PF13 with mixed bacterial communities, consisting of Gram-negative (Pseudomonas fluorescens and Klebsiella pneumoniae) and Gram-positive (Staphylococcus aureus and Enterococcus faecium) bacteria, was investigated to determine if this wild-type predator preferentially preys on certain bacteria and whether the presence of Gram-positive organisms influences its predation efficiency. In co-culture with P. fluorescens and K. pneumoniae, the cell counts (PFU/mL) of PF13 increased by 5.79 and 5.17 logs (48 h), respectively, while in the dual species assay (P. fluorescens, K. pneumoniae and PF13), the cell counts of PF13 increased by 1.95 logs (24 h). Using ethidium monoazide bromide quantitative polymerase chain reaction (EMA-qPCR), the concentration of PF13 increased by 1.25 to 3.62 logs in the co-culture experiments, by 1.41 to 5.05 logs in dual species cultures and by 2.65 logs in a polymicrobial culture. However, PF13 preferentially preyed on K. pneumoniae in the dual species and polymicrobial cultures, highlighting that the presence of Gram-positive bacteria did not affect the predation efficiency of PF13. This is significant as it implies that the predator can be applied in mixed microbial communities to target Gram-negative pathogens which may pose a health risk to patients, consumers or for the treatment of contaminated water. [ABSTRACT FROM AUTHOR]

The complete genome sequence of Bdellovibrio bacteriovorus 109J, a well-studied laboratory strain of predatory bacteria, first determined in 2014. Here we report an improved complete genome sequence of B. bacteriovorus 109J, incorporating 16 assembly and 87 nucleotide corrections. This revised genome will be helpful to studies on the predatory bacteria., Competing Interests: The authors declare no conflict of interest.

Bdellovibrio bacteriovorus is one of the predatory bacteria; therefore, it can act as a novel “living antibiotic,” unlike the current antibiotics. Here the predation of Escherichia coli by B. bacteriovorus was inhibited in the presence of Pseudomonas aeruginosa. This study investigated whether P. aeruginosa-induced predation inhibition is associated with bacterial quorum sensing (QS). Each las, rhl, or pqs QS mutant in P. aeruginosa was used to check the predatory activity of E. coli cells using B. bacteriovorus. As a result, the predatory activity of B. bacteriovorus increased in a mutant pqs QS system, whereas wild-type PA14 inhibited the predatory activity. Moreover, the addition of 4-hydroxy-2-heptylquinoline (HHQ) or the analog triggered the low predatory activity of B. bacteriovorus and killed B. bacteriovorus cells. Therefore, a defensive action of P. aeruginosa against B. bacteriovorus is activated by the pqs QS system, which produces some quinolone compounds such as HHQ.

Bdellovibrio bacteriovorus is one of the predatory bacteria; therefore, it can act as a novel "living antibiotic," unlike the current antibiotics. Here the predation of Escherichia coli by B. bacteriovorus was inhibited in the presence of Pseudomonas aeruginosa. This study investigated whether P. aeruginosa -induced predation inhibition is associated with bacterial quorum sensing (QS). Each las , rhl , or pqs QS mutant in P. aeruginosa was used to check the predatory activity of E. coli cells using B. bacteriovorus. As a result, the predatory activity of B. bacteriovorus increased in a mutant pqs QS system, whereas wild-type PA14 inhibited the predatory activity. Moreover, the addition of 4-hydroxy-2-heptylquinoline (HHQ) or the analog triggered the low predatory activity of B. bacteriovorus and killed B. bacteriovorus cells. Therefore, a defensive action of P. aeruginosa against B. bacteriovorus is activated by the pqs QS system, which produces some quinolone compounds such as HHQ. [ABSTRACT FROM AUTHOR]

Bdellovibrio bacteriovorus is a small Deltaproteobacterium which, since its discovery, has distinguished itself for the unique ability to prey on other Gram-negative bacteria. The studies on this particular "predatory bacterium", have gained momentum in response to the rising problem of antibiotic resistance, because it could be applied as a potential probiotic and antibiotic agent. Hereby, we present recent advances in the study of B. bacteriovorus, comprehending fundamental aspects of its biology, obligatory intracellular life cycle, predation resistance, and potential applications. Furthermore, we discuss studies that pave the road towards the use of B. bacteriovorus as a "living antibiotic" in human therapy, focussing on its interaction with biofilms, the host immune response, predation susceptibility and in vivo application models. The available data imply that it will be possible to upgrade this predator bacterium from a predominantly academic interest to an instrument that could confront antibiotic resistant infections. [ABSTRACT FROM AUTHOR]

The predatory bacterium Bdellovibrio bacteriovorus HW1 and bacteriophage ABTW1 were applied simultaneously, with solar disinfection (SODIS), for the reduction of Acinetobacter baumannii in environmental water sources. The B. bacteriovorus HW1 and ABTW1 dosing protocol was optimised, with results indicating that the simultaneous application of B. bacteriovorus HW1 and ABTW1 at 0 h resulted in the greatest log reduction (5.66 and 3.97 log based on CFU/mL and GC/mL, respectively) in A. baumannii over the 96 h co-culture period. Similar log reductions (5.24 and 3.85 log reduction based on CFU/mL and GC/mL, respectively) were however, obtained for the initial dosing of B. bacteriovorus at 0 h, and addition of bacteriophage ABTW1 at 24 h. A lyophilised bacteriophage formulation (0.5 M sucrose and 1% gelatine) was then compared to the standard liquid bacteriophage inoculum, with B. bacteriovorus , in upscaled volume trials. However, as similar A. baumannii log reduction results were obtained, the standard bacteriophage liquid inoculum, with B. bacteriovorus , was assessed as a pre-treatment to 6 h SODIS. The B. bacteriovorus HW1-bacteriophage ABTW1 combination as well as the B. bacteriovorus HW1 only pre-treatment, both with SODIS, significantly reduced the A. baumannii cell counts (8.32 and 8.38 log, respectively) to below detection limit (BDL). Molecular analysis however, showed that A. baumannii persisted (∼ 3 log reduction) after the application of all treatment protocols. It is thus recommended that the efficiency of the B. bacteriovorus -bacteriophage combination is further optimised to improve the lytic effect of both biological control agents, particularly the bacteriophage, on their prey. [Display omitted] [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The survival, proliferation, and epidemic spread of Acinetobacter baumannii (A. baumannii) in hospital settings is associated with several characteristics, including resistance to many commercially available antibiotics as well as the expression of multiple virulence mechanisms. This severely limits therapeutic options, with increased mortality and morbidity rates recorded worldwide. The World Health Organisation, thus, recognises A. baumannii as one of the critical pathogens that need to be prioritised for the development of new antibiotics or treatment. The current review will thus provide a brief overview of the antibiotic resistance and virulence mechanisms associated with A. baumannii’s “persist and resist strategy”. Thereafter, the potential of biological control agents including secondary metabolites such as biosurfactants [lipopeptides (surfactin and serrawettin) and glycolipids (rhamnolipid)] as well as predatory bacteria (Bdellovibrio bacteriovorus) and bacteriophages to directly target A. baumannii, will be discussed in terms of their in vitro and in vivo activity. In addition, limitations and corresponding mitigations strategies will be outlined, including curtailing resistance development using combination therapies, product stabilisation, and large-scale (up-scaling) production.

The interaction of Bdellovibrio bacteriovorus PF13 with mixed bacterial communities, consisting of Gram-negative (Pseudomonas fluorescens and Klebsiella pneumoniae) and Gram-positive (Staphylococcus aureus and Enterococcus faecium) bacteria, was investigated to determine if this wild-type predator preferentially preys on certain bacteria and whether the presence of Gram-positive organisms influences its predation efficiency. In co-culture with P. fluorescens and K. pneumoniae, the cell counts (PFU/mL) of PF13 increased by 5.79 and 5.17 logs (48 h), respectively, while in the dual species assay (P. fluorescens, K. pneumoniae and PF13), the cell counts of PF13 increased by 1.95 logs (24 h). Using ethidium monoazide bromide quantitative polymerase chain reaction (EMA-qPCR), the concentration of PF13 increased by 1.25 to 3.62 logs in the co-culture experiments, by 1.41 to 5.05 logs in dual species cultures and by 2.65 logs in a polymicrobial culture. However, PF13 preferentially preyed on K. pneumoniae in the dual species and polymicrobial cultures, highlighting that the presence of Gram-positive bacteria did not affect the predation efficiency of PF13. This is significant as it implies that the predator can be applied in mixed microbial communities to target Gram-negative pathogens which may pose a health risk to patients, consumers or for the treatment of contaminated water.

Bdellovibrio bacteriovorus 109J is a predatory bacterium which lives by predating on other Gram-negative bacteria to obtain the nutrients it needs for replication and survival. Here, we evaluated the effects two classes of bacterial signaling molecules (acyl homoserine lactones (AHLs) and diffusible signaling factor (DSF)) have on B. bacteriovorus 109J behavior and viability. While AHLs had a non-significant impact on predation rates, DSF considerably delayed predation and bdelloplast lysis. Subsequent experiments showed that 50 μM DSF also reduced the motility of attack-phase B. bacteriovorus 109J cells by 50% (38.2 ± 14.9 vs. 17 ± 8.9 μm/s). Transcriptomic analyses found that DSF caused genome-wide changes in B. bacteriovorus 109J gene expression patterns during both the attack and intraperiplasmic phases, including the significant downregulation of the flagellum assembly genes and numerous serine protease genes. While the former accounts for the reduced speeds observed, the latter was confirmed experimentally with 50 μM DSF completely blocking protease secretion from attack-phase cells. Additional experiments found that 30% of the total cellular ATP was released into the supernatant when B. bacteriovorus 109J was exposed to 200 μM DSF, implying that this QS molecule negatively impacts membrane integrity. [ABSTRACT FROM AUTHOR]

In bacteria, cell poles function as subcellular compartments where proteins localize during specific lifecycle stages, orchestrated by polar "hub" proteins. Whereas most described bacteria inherit an "old" pole from the mother cell and a "new" pole from cell division, generating cell asymmetry at birth, non-binary division poses challenges for establishing cell polarity, particularly for daughter cells inheriting only new poles. We investigated polarity dynamics in the obligate predatory bacterium Bdellovibrio bacteriovorus , proliferating through filamentous growth followed by non-binary division within prey bacteria. Monitoring the subcellular localization of two proteins known as polar hubs in other species, RomR and DivIVA, revealed RomR as an early polarity marker in B. bacteriovorus . RomR already marks the future anterior poles of the progeny during the predator's growth phase, during a precise period closely following the onset of divisome assembly and the end of chromosome segregation. In contrast to RomR's stable unipolar localization in the progeny, DivIVA exhibits a dynamic pole-to-pole localization. This behavior changes shortly before the division of the elongated predator cell, where DivIVA accumulates at all septa and both poles. In vivo protein interaction networks for DivIVA and RomR, mapped through endogenous miniTurbo-based proximity labeling, further underscore their distinct roles in cell polarization and reinforce the importance of the anterior "invasive" cell pole in prey-predator interactions. Our work also emphasizes the precise spatiotemporal order of cellular processes underlying B. bacteriovorus proliferation, offering insights into the subcellular organization of bacteria with filamentous growth and non-binary division.IMPORTANCEIn bacteria, cell poles are crucial areas where "hub" proteins orchestrate lifecycle events through interactions with multiple partners at specific times. While most bacteria exhibit one "old" and one "new" pole, inherited from the previous division event, setting polar identity poses challenges in bacteria with non-binary division. This study explores polar proteins in the predatory bacterium Bdellovibrio bacteriovorus , which undergoes filamentous growth followed by non-binary division inside another bacterium. Our research reveals distinct localization dynamics of the polar proteins RomR and DivIVA, highlighting RomR as an early "hub" marking polar identity in the filamentous mother cell. Using miniTurbo-based proximity labeling, we uncovered their unique protein networks. Overall, our work provides new insights into the cell polarity in non-binary dividing bacteria., Competing Interests: The authors declare no conflict of interest.

Bdellovibrio bacteriovorus is a predatory bacterium that feeds upon and proliferates inside other Gram-negative bacteria. Upon entry into the periplasmic space of the prey envelope, B. bacteriovorus initiates an exquisite developmental program in which it digests the host resources and grows as a filament, which eventually divides in a non-binary manner, releasing a variable number of daughter cells. The progeny then escape from the prey ghost to encounter new victims and resume the predation cycle. Owing to its unique biology, B. bacteriovorus undoubtedly represents an attractive model to unravel novel mechanisms of bacterial cell cycle control and cellular organization. Yet, the molecular factors behind the sophisticated lifestyle of this micro-predator are still mysterious. In particular, the spatiotemporal dynamics of proteins that control key cellular processes such as transmission of the genetic information, cell growth and division remain largely unexplored. In this Perspective article, I highlight outstanding fundamental questions related to these aspects and arising from the original biology of this bacterium. I also discuss available insights and potential cell biology approaches based on quantitative live imaging techniques, in combination with bacterial genetics and biochemistry, to shed light on the intracellular organization of B. bacteriovorus in space and time.

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. [ABSTRACT FROM AUTHOR]

Bdellovibrio bacteriovorus is a small Gram-negative bacterium and an obligate predator of other Gram-negative bacteria. Prey resistance to B. bacteriovorus attack is rare and transient. This consideration together with its safety and low immunogenicity makes B. bacteriovorus a valid alternative to antibiotics, especially in the treatment of multidrug resistant pathogens. In this study we developed a novel technique to estimate B. bacteriovorus sensitivity against antibiotics in order to make feasible the development and testing of co-therapies with antibiotics that would increase its antimicrobial efficacy and at the same time reduce the development of drug resistance. Results from tests performed with this technique show that among all tested antibiotics, trimethoprim has the lowest antimicrobial effect on B. bacteriovorus. Additional experiments revealed that the mechanism of trimethoprim resistance in B. bacteriovorus depends on the low affinity of this compound for the B. bacteriovorus dihydrofolate reductase (Bd DHFR). [ABSTRACT FROM AUTHOR]

Summary: Bdellovibrio bacteriovorus is a predator micro‐organism towards other Gram‐negative bacteria. We tested B. bacteriovorus to control Escherichia coli growth on chicken slices and canned beef. Moreover, we analysed B. bacteriovorus's lytic ability on eight toxigenic or multidrug‐resistant E. coli strains. In chicken slices, the predator induced the highest prey reduction (4.3 log) respect to control at 6 h. In canned beef, the predator induced the highest prey reduction (2.1 log) respect to control at 6 h. Moreover, B. bacteriovorus showed lytic ability towards all tested E. coli strains. B. bacteriovorus could control E. coli and other pathogenic and spoilage bacteria in those meat‐based foods that have a shelf life <10 days. It could integrate modified atmosphere packaging (MAP) to prolong the shelf life and improve the safety of pre‐packed fresh meat, meat preparations and meat products. In future applications on meat‐based foods, B. bacteriovorus could also minimise the use of additives. [ABSTRACT FROM AUTHOR]

Bdellovibrio bacteriovorus is a predatory bacterium that feeds upon and proliferates inside other Gram-negative bacteria. Upon entry into the periplasmic space of the prey envelope, B. bacteriovorus initiates an exquisite developmental program in which it digests the host resources and grows as a filament, which eventually divides in a non-binary manner, releasing a variable number of daughter cells. The progeny then escape from the prey ghost to encounter new victims and resume the predation cycle. Owing to its unique biology, B. bacteriovorus undoubtedly represents an attractive model to unravel novel mechanisms of bacterial cell cycle control and cellular organization. Yet, the molecular factors behind the sophisticated lifestyle of this micro-predator are still mysterious. In particular, the spatiotemporal dynamics of proteins that control key cellular processes such as transmission of the genetic information, cell growth and division remain largely unexplored. In this Perspective article, I highlight outstanding fundamental questions related to these aspects and arising from the original biology of this bacterium. I also discuss available insights and potential cell biology approaches based on quantitative live imaging techniques, in combination with bacterial genetics and biochemistry, to shed light on the intracellular organization of B. bacteriovorus in space and time. [ABSTRACT FROM AUTHOR]

ABSTRACT Bdellovibrio bacteriovorus is a bacterial predator capable of killing and replicating inside most Gram-negative bacteria, including antibiotic-resistant pathogens. Despite growing interest in this organism as a potential therapeutic, many of its genes remain uncharacterized. Here, we perform a high-throughput genetic screen with B. bacteriovorus using transposon sequencing (Tn-seq) to explore the genetic requirements of predation. Two hundred one genes were deemed essential for growth in the absence of prey, whereas over 100 genes were found to be specifically required for predative growth on the human pathogens Vibrio cholerae and Escherichia coli in both planktonic and biofilm states. To further this work, we created an ordered-knockout library in B. bacteriovorus and developed new high-throughput techniques to characterize the mutants by their stage of deficiency in the predator life cycle. Using microscopy and flow cytometry, we confirmed 10 mutants defective in prey attachment and eight mutants defective in prey rounding. The majority of these genes are hypothetical and previously uncharacterized. Finally, we propose new nomenclature to group B. bacteriovorus mutants into classes based on their stage of predation defect. These results contribute to our basic understanding of bacterial predation and may be useful for harnessing B. bacteriovorus to kill harmful pathogens in the clinical setting. IMPORTANCE Bdellovibrio bacteriovorus is a predatory bacterium that can kill a wide range of Gram-negative bacteria, including many human pathogens. Given the global rise of antibiotic resistance and dearth of new antibiotics discovered in the past 30 years, this predator has potential as an alternative to traditional antibiotics. For many years, B. bacteriovorus research was hampered by a lack of genetic tools, and the genetic mechanisms of predation have only recently begun to be established. Here, we comprehensively identify and characterize predator genes required for killing bacterial prey, as well as genes that interfere in this process, which may allow us to design better therapeutic predators. Based on our study, we and other researchers may ultimately be able to genetically engineer strains that have improved killing rates, target specific species of prey, or preferentially target prey in the planktonic or biofilm state.

The increased interest in predatory bacteria due to their ability to kill antibiotic resistant bacteria has also highlighted their inherent plethora of hydrolytic enzymes, and their potential as natural sources of novel therapeutic agents and biotechnological tools. Here, we have identified and characterized a novel protease from the predatory bacterium Bdellovibrio bacteriovorus: BspE (Bdellovibrio elastase-like serine protease). Mapping preferential sites of proteolytic activity showed a single proteolytic cleavage site of native plasma IgA (pIgA) in the Fc-tail; as well as in the secretory component (SC) of secretory IgA (SIgA). Proteolysis of other native immunoglobulins and plasma proteins was either absent (IgG1 and 2, IgM, albumin and orosomucoid) or unspecific with multiple cleavage sites (IgG3 and 4, IgE, IgD). BspE displayed a broad activity against most amino acid bonds in shorter peptides and denatured proteins, with a slight preference for hydrolysis C-terminal of Y, V, F, S, L, R, P, E, and K. BspE autoproteolysis results in numerous cleavage products sustaining activity for more than 6 h. The enzymatic activity remained stable at pH 5.0–9.0 but was drastically reduced in the presence of MnCl2 and completely inhibited by ZnCl2. The hydrolysis of pIgA was subsequently utilized for the specific glycan characterization of the released pIgA Fc-tail (Asn459). Besides contributing to the basic knowledge of Bdellovibrio biology and proteases, we propose that BspE could be used as a potential tool to investigate the importance, and biological function of the pIgA Fc-tail.IMPORTANCEAntibodies are well-established as key components of the immune system, and the importance of antibody glycosylation is steadily gaining recognition. Modifications of antibodies by glycosylation creates a vast repertoire of antibody glycovariants with distinctive and diverse functions in the immune system. Most of the available information regarding antibody glycosylation is based on studies with IgG, which have contributed greatly to the advance of therapeutic antibody treatments. However, much is still unknown regarding the importance of glycosylation and the Fc-structure for the remaining antibody classes. Such research has proven to be technically challenging and demonstrates a need for novel tools to facilitate such investigations. Here we have identified and characterized a novel protease from B. bacteriovorus, facilitating the study of plasma IgA by cleaving the Fc-tail, including the Asn459 N-glycan. This further highlights the potential of B. bacteriovorus as a source to identify potential novel biotechnological tools.

Endogenous endophthalmitis caused by Gram-negative bacteria is an intra-ocular infection that can rapidly progress to irreversible loss of vision. While most endophthalmitis isolates are susceptible to antibiotic therapy, the emergence of resistant bacteria necessitates alternative approaches to combat intraocular bacterial proliferation. In this study the ability of predatory bacteria to limit intraocular growth of Pseudomonas aeruginosa , Serratia marcescens , and Staphylococcus aureus was evaluated in a New Zealand white rabbit endophthalmitis prevention model. Predatory bacteria Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus were able to reduce proliferation of keratitis isolates of P. aeruginosa and to a lesser extent S. marcescens . However, it was not able to significantly reduce the number of intraocular S. aureus, which is not a productive prey for these predatory bacteria, suggesting that the inhibitory effect on P. aeruginosa and S. marcescens requires active predation rather than an antimicrobial immune response. Similarly, UV-inactivated B. bacteriovorus were unable to prevent proliferation of P. aeruginosa . Together, these data indicate in vivo inhibition of Gram-negative bacteria proliferation within the intra-ocular environment by predatory bacteria.

Bdellovibrio bacteriovorus is a predator bacterial species of the Deltaproteobacteria class that requires flagellum-mediated motility to initiate the parasitization of other gram-negative bacteria. The flagellum is capped by FliD, which polymerizes flagellin into a flagellar filament. FliD has been reported to function as a species-specific oligomer, such as a tetramer, a pentamer, or a hexamer, in members of the Gammaproteobacteria class. However, the oligomeric state and structural features of FliD from bacterial species outside the Gammaproteobacteria class are unknown. Based on structural and biochemical analyses, we report here that B. bacteriovorus FliD (bbFliD) forms a tetramer. bbFliD tetramerizes in a circular head-to-tail arrangement by inserting the D2 domain of one subunit into the concave surface of the second subunit generated between the D2 and D3 domains as observed in Serratia marcescens FliD. However, bbFliD adopts a more compact and flat oligomeric structure, which exhibits a more extended tetramerization interface flanked by two additional surfaces due to different intersubunit and interdomain organizations as well as an elongated loop. In conclusion, FliD from B. bacteriovorus , which belongs to the Deltaproteobacteria class, also produces a tetramer similar to FliD from Gammaproteobacterial species but adopts a unique species-specific oligomeric structure. • The crystal structure of Bdellovibrio bacteriovorus FliD (bbFliD) was determined. • bbFliD contains the D2 and D3 domains, which adopt β-cup structures. • bbFliD forms a tetramer, similar to Serratia marcescens FliD. • However, bbFliD exhibits a smaller tetramer size and a larger binding interface. [ABSTRACT FROM AUTHOR]

We evaluated the toxicity of surfactants against different predatory bacteria. Tests with Bdellovibrio bacteriovorus HD100 and SDS, an anionic surfactant, showed the predator was very sensitive; 0.02% SDS completely killed the predatory population (7-log loss; < 10 PFU/ml remaining) both when free-swimming or within the bdelloplast, i.e., intraperiplasmic. Similar results were also observed with B. bacteriovorus 109J and Peredibacter starrii. In contrast, none of the prey (E. coli, Klebsiella pneumoniae, Acinetobacter baumannii, or Pseudomonas sp. DSM 50906) viabilities were negatively affected by SDS. Triton X-100, a nonionic surfactant, was slightly less toxic towards B. bacteriovorus HD100 (viability loss of only 4-log), while two cationic surfactants, i.e., benzalkonium chloride (BZC) and cetyltrimethylammonium bromide (CTAB), were toxic towards both the predator and prey. Based on the above findings, we tested the potential use of SDS as a means to control predation. Addition of 0.02% SDS immediately halted predation based upon the prey bioluminescence, which leveled off and remained steady. This was confirmed using the predator viabilities; no predators were found in any of the samples where SDS was added. Consequently, low concentrations of SDS can be used as a simple means to control B. bacteriovorus HD100 activities. [ABSTRACT FROM AUTHOR]

Introduction: The oral microbiome is a complex community whose composition results from multiple interactions among different microorganisms and with the host. Predatory prokaryotes are recognized as important balancing factors in different ecosystems. Among them Bdellovibrio bacteriovorus received special attention for its strong predatory behaviour against many human pathogens in the absence of any toxic or pathogenic effect, so that it was proposed as a live probiotic/antibiotic agent. This work aimed to evaluate if B. bacteriovorus is detected from samples of human oral and dental biofilm. Materials and Methods: Samples of oral and dental biofilm were obtained from 20 adults of both sexes and processed for extraction of metagenomic DNA, to be used as templates for B. bacteriovorus-specific PCR reactions. Specificity of amplification products was confirmed by sequencing. Results: All 20 dental biofilm samples and 12 of 20 (60%) oral biofilm samples were resulted PCR positive. The sequences of 17 of 32 PCR products (53.1%) showed 100% identity with the reference sequence; the sequences of 11 of 32 PCR products (34.4%) showed ≥99% identity, while the remaining 4 products (12.5%) showed identities ranging between <99% and ≥97%. Conclusions: This is the first survey specifically reporting the presence of B. bacteriovorus in the human oral cavity and suggests that bacterial predation is a relevant balancing factor for the oral microbiota. Demonstration that B. bacteriovorus is able to colonize the oral cavity gives strength to proposals of its use as a probiotic/antibiotic in the prevention/treatment of selected oral diseases. [ABSTRACT FROM AUTHOR]

Background: This study evaluated the effects of topical administration of Bdellovibrio bacteriovorus HD100 on experimental periodontitis (EP) in rats.Methods: Thirty-two rats were divided into groups C (control), EP, C-HD100, and EP-HD100. At day 0, animals of groups EP and EP-HD100 received cotton ligatures around mandibular first molars (MFM). In groups C-HD100 and EP-HD100, 1 mL of suspensions containing B. bacteriovorus HD100 was topically administered in the subgingival region of MFMs at days 0, 3, and 7. Animals were euthanized at day 14. Gingival tissue, hemimandibles, and oral biofilm were collected. Data were statistically analyzed.Results: Group EP-HD100 presented greater bone volume and lower connective tissue attachment loss (CTAL) than group EP (P < 0.05). Group EP-HD100 presented greater proportions of Actinomyces and Streptococcus-like species and lower proportions of Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Fusobacterium polymorphum, Eikenella corrodens, Eubacterium nodatum, Campylobacter gracilis, Capnocytophaga sputigena, and Veillonella parvula-like species than group EP. Group EP-HD100 presented greater levels of osteoprotegerin and gene expression of interleukin (IL)-17, IL-10, and forkhead box P3 than group EP (P < 0.05).Conclusion: Topical use of B. bacteriovorus HD100 promotes a protective effect against alveolar bone loss and CTAL in rats with EP. [ABSTRACT FROM AUTHOR]