Predatory Bacteria Research Articles

Engineered Bdellovibrio bacteriovorus enhances antibiotic penetration and biofilm eradication.

Biofilms increase bacterial resistance to antibiotics, as conventional antibiotic doses are often ineffective at penetrating the biofilm matrix to eliminate bacteria. Recent research has shown that the Gram-negative predator bacterium Bdellovibrio bacteriovorus can penetrate Gram-positive bacterial biofilms during its predation phase and benefit from them without direct predation. Here, based on the penetration ability of B. bacteriovorus, we constructed antibiotic-loaded liposome-engineered B. bacteriovorus as a drug delivery strategy for biofilm-related diseases. As a "living antibiotic," B. bacteriovorus can prey on Gram-negative bacteria, penetrate biofilms, and disrupt their dense structure. During this process, the rapid movement of B. bacteriovorus enhances the delivery of antibiotic-loaded liposomes into the biofilm, promoting efficient antibiotic release and improving biofilm eradication. Our findings demonstrate that this engineered living antibiotic strategy significantly improves the control and removal of bacterial biofilms, accelerates the elimination of dental plaque, promotes wound healing, and holds promise as a novel platform for treating biofilm-related infections.

Unraveling the microecological mechanisms of phosphate-solubilizing Pseudomonas asiatica JP233 through metagenomics: insights into the roles of rhizosphere microbiota and predatory bacteria

The effects of phosphate-solubilizing bacteria (PSB) on plant productivity are high variable under field conditions. Soil phosphorus (P) levels are proposed to impact PSB performance. Furthermore, the effect of exogenous PSB on rhizosphere microbial community and their functions are largely unexplored. Our study examined how different P background and fertilization affected the performance of PSB Pseudomonas asiatica JP233. We further conducted metagenomic sequencing to assess its impact on rhizosphere microbiota and functions, with a focus on genes related to soil P cycling. We found that JP233 could enhance P solubilization and tomato growth to different extent in both high and low P soils, irrespective of P fertilization. It was particularly effective in high P soil without extra fertilization. JP233 altered the rhizosphere microbial community, boosting taxa known for plant growth promotion. It also changed soil gene profiling, enriching pathways related to secondary metabolite biosynthesis, amino acids, carbon metabolism, and other key processes. Particularly, JP233 increased the abundance of most P cycle genes and strengthened their interconnections. Populations of certain predatory bacteria increased after JP233 inoculation. Our findings provide valuable insights into PSB’s mechanisms for P solubilization and plant growth promotion, as well as potential adverse impacts of resident microbes on bioinoculants.

Predatory bacteria can intensify lung-injury in a multidrug-resistant Acinetobacter baumannii pneumonia model in rat.

Respiratory tract infection caused by antibiotic-resistant bacteria are one of the most important causes of death worldwide. Therefore, in this study, we investigated the possibility of using predatory bacteria to improve the Acinetobacter baumannii pneumonia model in rat. Multidrug-resistant (MDR) A. baumannii clinical strain was used to induce pneumonia. In addition to the sham and predator control group, three treatment groups (n = 5) were studied with colistin, Bdellovibrio bacteriovorus HD100, and combination of predator and antibiotics. Also, the colistin MIC value for B. bacteriovorus HD100 (8 μg/mL) was determined for the first time to our knowledge. Removal of excess endotoxin from the predator suspension was performed with the help of organic solvents before inoculation of rats. The most successful treatment was observed in the group treated with colistin followed by combined treatment. In the predator treatment group, the systemic spread of A. baumannii was lower than other treatment groups. However, treatment with predatory bacteria not only failed to reduce the pathogen load in the lungs to the same extent as the antibiotic treatment group, but also induced acute pulmonary and systemic inflammatory responses. Therefore, the rats showed the highest septic score (21.4 at 48 h) and did not survive more than 48 h. This is the first report of systemic complications of using B. bacteriovorus HD100 for infection control. According to our results, the effects of predatory bacteria in the in vivo environment are complex and many questions need to be answered before it can be introduced as a live antibiotic.

Myxobacteria from soil can substantially reduce the bacterial load in a wound infection model.

Myxobacteria are non-pathogenic, saprophytic, soil-dwelling predatory bacteria known for their antimicrobial potential. Many pathogenic bacteria form biofilms to protect themselves from antimicrobial agents and the immune system. This study has investigated the predatory activities of myxobacteria against pathogenic bacteria in biofilms. A total of 50 soil samples were collected in and around Cardiff, South Wales (UK). Using a baiting method with 6 prey organisms, 32 myxobacteria were isolated and identified by 16S rRNA sequencing, of which 18 were Myxococcus spp. and 14 were Corallococcus spp. Predation assays, biofilm inhibition and disruption assays, and a dynamic, polymicrobial wound biofilm model were used with live myxobacteria to assess efficacy of predation. Good activity in predation assays was observed against Escherichia coli, while Enterococcus faecalis was more recalcitrant to myxobacteria. Staphylococcus aureus and Citrobacter freundii were significantly (P<0.05) reduced in both biofilm inhibition and disruption assays compared to other pathogens. Considerable reductions (>3 log10 CFU) in the wound infection model were seen after 96h of incubation, particularly for C. freundii and E. coli. Using live predatory bacteria as an alternative therapeutic agent has received attention in the recent past to combat the problem of antimicrobial resistance. Myxobacteria isolated from soil using multiple prey organisms yielded diverse isolates, including strains which exhibited therapeutically promising activities in a variety of infection/biofilm assays.

Comparative genomics of obligate predatory bacteria belonging to phylum Bdellovibrionota highlights distribution and predicted functions of lineage-specific protein families.

Comparative genomics of predatory bacteria is important to understand their ecology and evolution and explore their potential to treat drug-resistant infections. We compared chromosomes of 18 obligate predators from phylum Bdellovibrionota (16 intraperiplasmic, two epibiotic) and 15 non-predatory bacteria. Phylogenetics of conserved single-copy genes and analysis of genome-wide average amino acid identity provide evidence for at least five Bdellovibrio species and support recent reclassifications of predatory taxa. To define shared and differential genome content, we grouped predicted protein sequences into gene clusters based on sequence similarity. Few gene clusters are shared by all 33 bacteria or all 18 predatory bacteria; however, we identified gene clusters conserved within lineages, such as intraperiplasmic Bdellovibrio, and not found in other bacteria. Many of these are predicted to function in cell envelope biogenesis, signal transduction, and other roles important for predatory lifestyles. Among intraperiplasmic Bdellovibrio, we detected high abundance of gene clusters predicted to encode transglycosylases, endopeptidases, and lysozymes, and we identified six gene clusters (amidase, L,D-transpeptidase, four transglycosylases) with evidence of recent gene duplication and gene family expansion. Focusing on peptidoglycan metabolism, we defined a suite of gene clusters that include peptidoglycan-degrading and -modifying enzymes and occur only in predatory bacteria, suggesting these proteins may have evolved activities specific to predation. Our analyses highlight key genome content differences between obligate predatory bacteria and non-predatory relatives and identify gene clusters that may encode enzymes adapted to predatory lifestyles. These lineage-specific proteins are strong candidates for functional characterization to clarify their role in predation.IMPORTANCEEvolution of predation as a bacterial lifestyle involves selective pressure on and adaptation of enzymes that contribute to killing and digestion of prey bacteria, in some cases from within the prey itself. Such enzymes are a hallmark of obligate predatory bacteria belonging to phylum Bdellovibrionota, which includes the well-studied predator Bdellovibrio. By comparing protein sequences of obligate predatory bacteria and their non-predatory relatives, we define key genome content differences that distinguish bacterial predators and identify lineage-specific enzymes that may have evolved unique activities due to selective pressures related to a predatory lifestyle. In addition to providing insights into the ecology and evolution of predatory bacteria, comparative genomics studies, like this, can inform efforts to develop predatory bacteria and/or their enzymes as potential biocontrol agents to combat drug-resistant bacterial infections.

Encapsulated Predatory Bacteria Efficiently Protect Potato Tubers from Soft Rot Disease.

Soft rot Pectobacteriaceae (SRP) are a group of destructive Gram-negative phytopathogens that can infect a wide range of plant hosts, including potatoes. There are no effective control agents available against SRP, making their management challenging. We have developed a novel approach to protect potato tubers against SRP. It makes use of encapsulated predatory Bdellovibrio bacteriovorus bacteria that, upon release from a polymeric carrier, prey upon SRP. We applied a carrageenan-trehalose-based formulation containing a B. bacteriovorus HD100 predator to prevent soft rot disease development in potato tubers, under various conditions. The dried formulation exhibited very high stability over an 18-month period at room temperature (∼25°C), in contrast to unencapsulated suspensions of the predator, in which viability decreased rapidly below detection level. The rehydrated formulation was as efficient as freshly grown unencapsulated predators and provided high protection in potted potato tubers, displaying an average of 50% reduction in disease parameters (e.g., tissue decay and disease index) under controlled conditions at 7 days postinoculation and planting. The protective effect provided by this formulation was maintained in longer-term trials (28 days) conducted in larger vessels within a net house under natural climate conditions, highlighting its potential for practical application in the field.

Bdellovibrio bacteriovorus Therapy, an Emerging Alternative to Antibiotics

: The increase in multi-drug resistant (MDR) pathogens and the decline in the number of new antibiotics in the production pipeline pose a serious threat to our ability to treat infectious diseases. In this new landscape, once treatable diseases are now potentially life-threatening. This impending danger requires that urgent attention should be given to developing alternative strategies for combating MDR bacteria. A novel alternative is the use of predatory bacteria, B. bacteriovorus spp, that naturally prey on Gram-negative bacteria, including MDR Enterobacteriaceae. B. bacteriovorus has been shown to be nonpathogenic in animal models and on human cell lines, supporting its feasibility to be used to treat infections in animals and possibly humans. This document reviews various aspects of B. bacteriovorus biology, including its unique life cycle, "predatory toolbox", prey range, and recent research advances exploring B. bacteriovorus as an antimicrobial agent, stepping towards its use in human therapy. We also discuss the advantages and limitations of using B. bacteriovorus therapy and the strategies to overcome these limitations.

Regulation of Antibiotic Resistance Genes on Agricultural Land Is Dependent on Both Choice of Organic Amendment and Prevalence of Predatory Bacteria.

Antibiotic resistance genes (ARGs) are widespread in the environment, and soils, specifically, are hotspots for microorganisms with inherent antibiotic resistance. Manure and sludge used as fertilizers in agricultural production have been shown to contain vast amounts of ARGs, and due to continued applications, ARGs accumulate in agricultural soils. Some soils, however, harbor a resilience capacity that could depend on specific soil properties, as well as the presence of predatory bacteria that are able to hydrolyse living bacteria, including bacteria of clinical importance. The objectives of this study were to (i) investigate if the antibiotic resistance profile of the soil microbiota could be differently affected by the addition of cow manure, chicken manure, and sludge, and (ii) investigate if the amendments had an effect on the presence of predatory bacteria. The three organic amendments were mixed separately with a field soil, divided into pots, and incubated in a greenhouse for 28 days. Droplet digital PCR (ddPCR) was used to quantify three ARGs, two predatory bacteria, and total number of bacteria. In this study, we demonstrated that the choice of organic amendment significantly affected the antibiotic resistance profile of soil, and promoted the growth of predatory bacteria, while the total number of bacteria was unaffected.

Diversity of Free-Living Amoebae in New Zealand Groundwater and Their Ability to Feed on Legionella pneumophila.

Free-living amoebae (FLA) are common in both natural and engineered freshwater ecosystems. They play important roles in biofilm control and contaminant removal through the predation of bacteria and other taxa. Bacterial predation by FLA is also thought to contribute to pathogen dispersal and infectious disease transmission in freshwater environments via the egestion of viable bacteria. Despite their importance in shaping freshwater microbial communities, the diversity and function of FLA in many freshwater ecosystems are poorly understood. In this study, we isolated and characterized FLA from two groundwater sites in Canterbury, New Zealand using microbiological, microscopic, and molecular techniques. Different methods for groundwater FLA isolation and enrichment were trialed and optimized. The ability of these isolated FLA to predate on human pathogen Legionella pneumophila was assessed. FLA were identified by 18S metagenomic amplicon sequencing. Our study showed that Acanthamoeba spp. (including A. polyphaga) and Vermamoeba veriformis were the main FLA species present in both groundwater sites examined. While most of the isolated FLA co-existed with L. pneumophila, the FLA populations in the L. pneumophila co-culture experiments predominantly consisted of A. polyphaga, Acanthamoeba spp., Naegleria spp., V. vermiformis, Paravahlkampfia spp., and Echinamoeba spp. These observations suggest that FLA may have the potential to act as reservoirs for L. pneumophila in Canterbury, New Zealand groundwater systems and could be introduced into the local drinking water infrastructure, where they may promote the survival, multiplication, and dissemination of Legionella. This research addresses an important gap in our understanding of FLA-mediated pathogen dispersal in freshwater ecosystems.

Aggregatibacter actinomycetemcomitans Dispersin B: The Quintessential Antibiofilm Enzyme.

The extracellular matrix of most bacterial biofilms contains polysaccharides, proteins, and nucleic acids. These biopolymers have been shown to mediate fundamental biofilm-related phenotypes including surface attachment, intercellular adhesion, and biocide resistance. Enzymes that degrade polymeric biofilm matrix components, including glycoside hydrolases, proteases, and nucleases, are useful tools for studying the structure and function of biofilm matrix components and are also being investigated as potential antibiofilm agents for clinical use. Dispersin B is a well-studied, broad-spectrum antibiofilm glycoside hydrolase produced by Aggregatibacter actinomycetemcomitans. Dispersin B degrades poly-N-acetylglucosamine, a biofilm matrix polysaccharide that mediates biofilm formation, stress tolerance, and biocide resistance in numerous Gram-negative and Gram-positive pathogens. Dispersin B has been shown to inhibit biofilm and pellicle formation; detach preformed biofilms; disaggregate bacterial flocs; sensitize preformed biofilms to detachment by enzymes, detergents, and metal chelators; and sensitize preformed biofilms to killing by antiseptics, antibiotics, bacteriophages, macrophages, and predatory bacteria. This review summarizes the results of nearly 100 in vitro and in vivo studies that have been carried out on dispersin B since its discovery 20 years ago. These include investigations into the biological function of the enzyme, its structure and mechanism of action, and its in vitro and in vivo antibiofilm activities against numerous bacterial species. Also discussed are potential clinical applications of dispersin B.

Increasing fish production in recirculating aquaculture system by integrating a biofloc-worm reactor for protein recovery

Aquaculture, producing half of global fish production, offers a high-quality protein source for humans. Improving nitrogen use efficiency (NUE) through microbial protein recovery is crucial for increasing fish production and reducing environmental footprint. However, the poor palatability and high moisture content of microbial protein make its utilization challenging. Here, a biofloc-worm reactor was integrated into a recirculating aquaculture system (BW_RAS) for the first time to convert microbial protein into Tubificidae (Oligochaeta) biomass, which was used as direct feed for culturing fish. Batch experiments indicated that an aeration rate of 0.132 m3 L -1 h -1 and a worm density of 0.3 g cm-2 on the carrier were optimal for microbial biomass growth and worm predation, respectively. Compared to the biofloc reactor-based recirculating aquaculture system (B_RAS), the BW_RAS improved water quality, NUE, and fish production by 17.1 % during a 120-day aquaculture period. The abundance of heterotrophic aerobic denitrifier Deinococcus in BW_RAS was one order of magnitude higher than in B_RAS, while heterotrophic bacteria Mycobacterium was more abundant in B_RAS. Denitrifiers cooperated with organic matter degraders and nitrogen assimilation bacteria for protein recovery and gaseous nitrogen loss while competing with predatory bacteria. Function prediction and qPCR indicated greater aerobic respiration, nitrate assimilation, nitrification (AOB-amoA), and denitrification (napA, nirK, nirS, nosZI), but lower fermentation in BWR compared to BR. This study demonstrated that BW_RAS increased microbial protein production and aerobic nitrogen cycling through ongoing worm predation, further enhancing fish production to a commercially viable level.

Postbiotic as Novel Alternative Agent or Adjuvant for the Common Antibiotic Utilized in the Food Industry.

Antibiotic resistance is a serious public health problem as it causes previously manageable diseases to become deadly infections that can cause serious disability or even death. Scientists are creating novel approaches and procedures that are essential for the treatment of infections and limiting the improper use of antibiotics in an effort to counter this rising risk. With a focus on the numerous postbiotic metabolites formed from the beneficial gut microorganisms, their potential antimicrobial actions, and recent associated advancements in the food and medical areas, this review presents an overview of the emerging ways to prevent antibiotic resistance. Presently, scientific literature confirms that plant-derived antimicrobials, RNA therapy, fecal microbiota transplantation, vaccines, nanoantibiotics, haemofiltration, predatory bacteria, immunotherapeutics, quorum-sensing inhibitors, phage therapies, and probiotics can be considered natural and efficient antibiotic alternative candidates. The investigations on appropriate probiotic strains have led to the characterization of specific metabolic byproducts of probiotics named postbiotics. Based on preclinical and clinical studies, postbiotics with their unique characteristics in terms of clinical (safe origin, without the potential spread of antibiotic resistance genes, unique and multiple antimicrobial action mechanisms), technological (stability and feasibility of largescale production), and economic (low production costs) aspects can be used as a novel alternative agent or adjuvant for the common antibiotics utilized in the production of animal-based foods. Postbiotic constituents may be a new approach for utilization in the pharmaceutical and food sectors for developing therapeutic treatments. Further metabolomics investigations are required to describe novel postbiotics and clinical trials are also required to define the sufficient dose and optimum administration frequency of postbiotics.

Predatory Bacteria in the Treatment of Infectious Diseases and Beyond.

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.

Flagellar stator genes control a trophic shift from obligate to facultative predation and biofilm formation in a bacterial predator.

The ability of bacteria to form biofilms is a central research theme in biology, medicine, and the environment. We show that cultures of the obligate (host-dependent) "solitary" predatory bacterium Bdellovibrio bacteriovorus, which cannot replicate without prey, can use various genetic routes to spontaneously yield host-independent (H-I) variants that grow axenically (as a single species, in the absence of prey) and exhibit various surface attachment phenotypes, including biofilm formation. These routes include single mutations in flagellar stator genes that affect biofilm formation, provoke motor instability and large motility defects, and disrupt cyclic-di-GMP intracellular signaling. H-I strains also exhibit reduced predatory efficiency in suspension but high efficiency in prey biofilms. These changes override the requirements for prey, enabling a shift from obligate to facultative predation, with potential consequences on community dynamics.

Bdellovibrio’s prey-independent lifestyle is fueled by amino acids as a carbon source

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.

Improved complete genome sequence of Bdellovibrio bacteriovorus 109J, a widely studied laboratory strain of predatory bacteria.

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.

Surface Microstructure Drives Biofilm Formation and Biofouling of Graphene Oxide Membranes in Practical Water Treatment.

Significant progress has been made previously in the research and development of graphene oxide (GO) membranes for water purification, but their biofouling behavior remains poorly understood. In this study, we investigated the biofilm formation and biofouling of GO membranes with different surface microstructures in the context of filtering natural surface water and for an extended operation period (110 days). The results showed that the relatively hydrophilic and smooth Fe(OH)3/GO membrane shaped a thin and spatially heterogeneous biofilm with high stable flux. However, the ability to simultaneously mitigate biofilm formation and reduce biofouling was not observed in the weakly hydrophilic and wrinkled Fe/GO and H-Fe(OH)3/GO membranes. Microbial analyses revealed that the hydrophilicity and roughness distinguished the bacterial communities and metabolic functions. The organic matter-degrading and predatory bacteria were more adapted to hydrophilic and smooth GO surfaces. These functional taxa were involved in the degradation of extracellular polymeric substances (EPS), and improved biofilm heterogeneity. In contrast, the weakly hydrophilic and wrinkled GO surfaces had reduced biodiversity, while unexpectedly boosting the proliferation of EPS-secreting bacteria, resulting in increased biofilm formation and aggravated biofouling. Moreover, all GO membranes achieved sustainable water purification during the entire operating period.

MyxoPortal: a database of myxobacterial genomic features.

Myxobacteria are predatory bacteria with antimicrobial activity, utilizing complex mechanisms to kill their prey and assimilate their macromolecules. Having large genomes encoding hundreds of secondary metabolites, hydrolytic enzymes and antimicrobial peptides, these organisms are widely studied for their antibiotic potential. MyxoPortal is a comprehensive genomic database hosting 262 genomes of myxobacterial strains. Datasets included provide genome annotations with gene locations, functions, amino acids and nucleotide sequences, allowing analysis of evolutionary and taxonomical relationships between strains and genes. Biosynthetic gene clusters are identified by AntiSMASH, and dbAMP-generated antimicrobial peptide sequences are included as a resource for novel antimicrobial discoveries, while curated datasets of CRISPR/Cas genes, regulatory protein sequences, and phage associated genes give useful insights into each strain's biological properties. MyxoPortal is an intuitive open-source database that brings together application-oriented genomic features that can be used in taxonomy, evolution, predation and antimicrobial research. MyxoPortal can be accessed at http://dicsoft1.physics.iisc.ac.in/MyxoPortal/. Database URL: http://dicsoft1.physics.iisc.ac.in/MyxoPortal/. Graphical Abstract.

Sludge biolysis pretreatment to reduce antibiotic resistance genes (ARGs): Insight into the relationship between potential ARGs hosts and BALOs' preferred prey

As an important reservoir of antibiotic resistance genes (ARGs), the sludge discharged from wastewater treatment plants is the key intermediate for ARG transport into the environment. Bdellovibrio-and-like organisms (BALOs) are predatory bacteria that are expected to attack antibiotic-resistant bacteria (ARB). In this study, the screened BALOs (C3 & D15) were mixed with the sludge for biolysis to achieve the satisfying removal efficiencies of six tet genes, two sul genes, and one mobile genetic element (intl 1). Among them, tet(Q) demonstrated the highest reduction rate in relative abundance at 87.3 ± 1.0 %, while tet(X) displayed the lowest of 11.7 ± 0.2 %. The microorganisms, including Longilinea, Methanobacterium, Acetobacterium, Sulfurimonas, allobaculum, Gaiella, AAP99, Ellin6067, Rhodoferax, Ferruginibacter and Thermomonas, were expected to play a dual role in the reduction of ARGs by serving as ARB and BALOs' preferred prey. Meanwhile, BALOs consortium improved ARGs reduction efficiency via the expansion of the prey profile. Additionally, BALOs decreased the relative abundance of not only pathogens (Shinella, Rickettsia, Burkholderia, Acinetobacter, Aeromonas, Clostridium, Klebsiella and Pseudomonas), but also the ARGs' host pathogens (Mycobacterium, Plesiocystis, Burkholderia, and Bacteroides). Therefore, the application of BALOs for sludge biolysis are promising to decrease the sludge's public health risks via limiting the spread of ARGs and pathogens into the environment.

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

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