Anaerobic Bacteria Research Articles

Evaluation of VITEK MS Version 3.0 MALDI-TOF for the identification of anaerobes, mycobacteria, Nocardia, and moulds

PurposeThe identification of anaerobes, Mycobacterium and Nocardia species, and moulds by MALDI-TOF-MS remains a challenge. This study aimed to evaluate the performance of MALDI-TOF in the identification of these organisms. MethodsA total of 382 strains, comprising 128 (33.5 %) anaerobes, 126(33.0 %) mycobacterial, 113(29.6 %), mycelial fungi, and 15(3.9 %) Nocardia species were evaluated by VITEK MS Version 3.0. The results were compared with the identification of the isolates by DNA sequence analysis. The DNA sequences used for analysis were the 16S rRNA for anaerobic bacteria, hsp65 gene for mycobacteria, whereas both 16S rRNA and hsp65 gene for Nocardia species, and internal transcribed spacer (ITS) and 28S rRNA gene's D1/D2 regions of fungi. ResultsThe VITEK-MS accurately identified 78.3 % (299/382) of the strains at the species, and 9.4 % (36/382) at the genus level. Misidentifications were observed in 3.9 % (15/382) isolates. Of isolates tested, 8.4 % (32/382) were not identified by the system, and 7.06 % (27/382) were not included in the IVD database. ConclusionAn upgraded VITEK MS V3.0 database provides reasonably accurate and rapid identification of clinically relevant anaerobes, mycobacteria, Nocardia species, and moulds to the species level.

Microbial-mediated metal(loid) immobilization in mulch-covered tailings

Mulch coverage of mining tailings can create anaerobic conditions and consequently establish an anoxic environment that promotes the metabolic processes of anaerobic microorganisms. This anoxic environment has the potential to decrease heavy metal mobility and bioavailability. While tailings exposed to sunlight have been extensively studied, research on the effects of microbial-mediated geochemical cycling of heavy metals in mulch-covered tailings is scarce. This study aimed to examine the effects of mulch coverage-induced alterations in the structures of tailing microbial communities on the biogeochemical processes associated with heavy metals. Mulch coverage significantly reduced the pH of the tailings and the tailings exhibited heavy metal bioavailability. Random forest analysis demonstrated that mulch coverage-induced changes in the As/Cd-contaminated fractions and nutrients (total organic carbon and total nitrogen) were the most crucial predictors of microbial diversity and ecological clusters in the tailings. Notably, different from direct metal(loid) immobilization, mulch coverage can facilitate heavy metal immobilization in tailings by promoting microbial-mediated Fe, S, and As reduction. Overall, this study demonstrated that mulch coverage of tailings contributed to a reduction in heavy metal mobilization, which can be attributed to shifts in microbial-mediated Fe, S, and As reduction processes.The study provides valuable insights into the potential of mulch coverage as a remediation strategy and underscores the importance of microbial-mediated processes in managing heavy metal pollution in tailing systems.

In vivo detection of endogenous toxic phenolic compounds of intestine

Phenol and p-cresol are two common toxic small molecules related to various diseases. Existing reports confirmed that high L-tyrosine in the daily diet can increase the concentration of phenolic compounds in blood and urine. L-tyrosine is a common component of protein-rich foods. Some anaerobic bacteria in the gut can convert non-toxic l-tyrosine into these two toxic phenolic compounds, phenol and p-cresol. Existing methods have been constructed for measuring the concentration of phenolic compound in feces. However, there is still a lack of direct visual evidence to measure the phenolic compounds in the intestine. In this study, we aimed to construct a whole-cell biosensor for phenolic compounds detection based on the dmpR, the regulator from the phenol metabolism cluster. The commensal bacterium Citrobacter amalonaticus PS01 was selected and used as the chassis. Compared with the biosensor based on ECN1917, the biosensor PS01[dmpR] could better implant into the mouse gut through gavage and showed a higher sensitive to phenolic compound. And the concentration of phenolic compounds in the intestines could be observed with the help of in vivo imaging system using PS01[dmpR]. This paper demonstrated endogenous phenol synthesis in the gut and the strategy of using commensal bacteria to construct whole-cell biosensors for detecting small molecule compounds in the intestines.

Complete genome sequence of Bifidobacterium animalis B01.

Bifidobacterium, a class of strictly anaerobic Gram-positive bacteria, existed mainly in the gut of humans and many mammals. Here, we report the complete genome sequence of Bifidobacterium animalis B01, whose genome length is 1,935,423 bp with a GC content of 60.49%.

Anaerobic bacteria in chronic wounds: Roles in disease, infection and treatment failure.

Infection is among the most common factors that impede wound healing, yet standard treatments routinely fail to resolve chronic wound infections. The chronic wound environment is largely hypoxic/anoxic, and wounds are predominantly colonised by facultative and obligate anaerobic bacteria. Oxygen (O2) limitation is an underappreciated driver of microbiota composition and behaviour in chronic wounds. In this perspective article, we examine how anaerobic bacteria and their distinct physiologies support persistent, antibiotic-recalcitrant infections. We describe the anaerobic energy metabolisms bacteria rely on for long-term survival in the wound environment, and why many antibiotics become less effective under hypoxic conditions. We also discuss obligate anaerobes, which are among the most prevalent taxa to colonise chronic wounds, yet their potential roles in influencing the microbial community and wound healing have been overlooked. All of the most common obligate anaerobes found in chronic wounds are opportunistic pathogens. We consider how these organisms persist in the wound environment and interface with host physiology to hinder wound healing processes or promote chronic inflammation. Finally, we apply our understanding of anaerobic physiologies to evaluate current treatment practices and to propose new strategies for treating chronic wound infections.

Commensal-derived short-chain fatty acids disrupt lipid membrane homeostasis in Staphylococcus aureus.

The role of commensal anaerobic bacteria in chronic respiratory infections is unclear, yet they can exist in abundances comparable to canonical pathogens in vivo. Their contributions to the metabolic landscape of the host environment may influence pathogen behavior by competing for nutrients and creating inhospitable conditions via toxic metabolites. Here, we reveal a mechanism by which the anaerobe-derived short chain fatty acids (SCFAs) propionate and butyrate negatively affect Staphylococcus aureus physiology by disrupting branched chain fatty acid (BCFA) metabolism. In turn, BCFA impairment results in impaired growth, diminished expression of the agr quorum sensing system, as well as increased sensitivity to membrane-targeting antimicrobials. Altered BCFA metabolism also reduces S. aureus fitness in competition with Pseudomonas aeruginosa, suggesting that airway microbiome composition and the metabolites they produce and exchange directly impact pathogen succession over time. The pleiotropic effects of these SCFAs on S. aureus fitness and their ubiquity as metabolites in animals also suggests that they may be effective as sensitizers to traditional antimicrobial agents when used in combination.

Insights into the biocorrosion of Q235A steel influenced by the electron transfer process between iron and methanogenic microbiota

Anaerobic microbiologically influenced corrosion (MIC) of Fe (0) metals causes great harm to the environment and economy, which depends on the key electron transfer process between anaerobic microorganisms and Fe (0) metals. However, the key electron transfer process in microbiota dominating MIC remains unclear, especially for methanogenic microbiota wildly distributed in the environment. Herein, three different methanogenic microbiota (Methanothrix, Methanospirillum, and Methanobacterium) were acclimated to systematically investigate electron transfer pathways on corroding Q235A steel coupons. Results indicated that microbiota dominated by Methanothrix, Methanospirillum, or Methanobacterium accelerated the steel corrosion mainly through direct electron transfer (DET) pathway, H2 mediated electron transfer (HMET) pathway, and combined DET and HMET pathways, respectively. Compared with Methanospirillum dominant microbiota, Methanothrix or Methanobacterium dominant microbiota caused more methane production, higher weight loss, corrosion pits with larger areas, higher corrosion depth, and smaller corrosion pits density. Such results reflected that the DET process between microbiota and Fe (0) metals decided the biocorrosion degree and behavior of Fe (0) metals. This study insightfully elucidates the mechanisms of methanogenic microbiota on corroding steels, in turn providing new insights for anti-corrosion motives.

Enhancing methane production from corn straw via illumination-assisted Fe3O4/g-C3N4 nanocomposite in anaerobic digestion

This study proposes a novel anaerobic digestion (AD) strategy combining recyclable photoactivated nanomaterials with illumination to enhance electronic transfer for anaerobic microorganisms. Results showed that 7000 Lux illumination increased methane production yield and rate. Incorporating Fe3O4 into graphite carbon nitride (g-C3N4) created a recyclable Fe3O4/g-C3N4 (FG) nanocomposite with improved light absorption, conductivity, redox properties, and methane promotion. The highest methane yield from corn straw was achieved with 7000 Lux and 1.5 g/L FG nanocomposite, 22.6% higher than the dark control. The AD system exhibited increased adenosine triphosphate content, improved redox performance, reduced electron transfer resistance, and higher photocurrent intensity. These improvements bolstered the microorganisms and key genes involved in hydrolysis and acidification, which in turn optimized the acetoclastic pathway. Furthermore, this strategy promoted microorganisms associated with direct interspecies electron transfer, fostering a favorable environment for methanogenic activities, paving the way for future anaerobic reactor developments.

Effectiveness of Biological Approaches for Removing Persistent Organic Pollutants from Wastewater: A Mini-Review.

Persistent organic pollutants (POPs), including organochlorine pesticides, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzo-p-furans, pose significant hazards to the environment and living organisms. This concise review aims to consolidate knowledge on the biological processes involved in removing POPs from wastewater, an area less explored compared to conventional physico-chemical methods. The focus is on the potential of various aerobic and anaerobic microorganisms, fungi, and bacteria for efficient bioremediation, mitigating or eradicating the deleterious effects of these chemicals. The review scrutinizes individual bacterial strains and mixed cultures engaged in breaking down persistent organic pollutants in water, highlighting promising results from laboratory investigations that could be scaled for practical applications. The review concludes by underscoring the opportunities for exploring and advancing more sophisticated bioremediation techniques and optimized bioreactors. The ultimate goal is to enhance the efficiency of microbial-based strategies, implicitly reducing the environmental impact of persistent chemicals.

Homogeneous HER electrocatalysis using monothiolate ligand-based {FeS} complexes: A review

Renewable energy sources are considered as efficient, low cost and sustainable energy systems with the capability of achieving a decarbonized world. Of particular interest has been the use of hydrogen as an affordable and clean energy for achieving a sustainable (renewable) energy economy. The hydrogenases discovered almost 90 years ago and found in green algae and anaerobic bacteria, are known to be highly efficient biological catalysts for hydrogen evolution in nature. Out of the three types of hydrogenases ([FeFe], [NiFe] and [Fe] only) the [FeFe] hydrogenases have drawn particular attention due to their higher activity/efficiency and their ability to reversibly reduce protons to dihydrogen. Several models of the [FeFe] hydrogenase enzymes have been reported over the last three decades. This review article mainly describes in details the characteristics of mono- and di-nuclear {FeS} complexes with monothiolates (-SR) as the bridging ligands that have been reported as electrocatalysts for the hydrogen evolution reaction (HER).

Cardiorespiratory and aerobic demands of squat exercise

Squatting, a traditional resistance exercise classified as strength training, relies on anaerobic pathways, but its aerobic aspects remain unclear. We examined heart rate and oxygen demand during squats, exploring variations across different strength statuses. It fills gaps in understanding the cardiorespiratory effects of squatting, especially during multiple sets. Twenty-two young healthy resistance trained men (age: 28 ± 4 years) participated. Maximal oxygen consumption (V̇O2max) and 1 repetition maximum (RM) of squat were measured. Participants performed 5 sets of squat exercises at 65% of 1RM for 10 repetitions with 3-min rest intervals. Heart rate and pulmonary gas exchange were measured during the squat exercise. Participants were divided into high strength (HS; upper 50%) and low strength (LS; lower 50%) groups based on a median split of their 1 RM squat values (normalized to their body weight). During 5 sets of squat exercise, oxygen consumption (V̇O2) increased up to 47.8 ± 8.9 ml/kg/min, corresponding to 100.6% of predetermined V̇O2max. The HS group achieved a greater highest point of V̇O2 in relation to V̇O2max than the LS group (108.0 vs. 93.7%). During the exercise intervals, V̇O2 exceeded V̇CO2, while during the rest intervals, V̇CO2 surpassed V̇O2. Our findings suggest that the oxygen demand during squatting is notably substantial, which may vary according to the training status.

Mechanistic Basis for the Translation Inhibition of Cutibacterium acnes by Clindamycin

Inflammation and the Gram-positive anaerobic bacterium Cutibacterium acnes, which is implicated in acne pathogenesis and pilosebaceous unit inflammation, are the main targets of antibiotic-based therapy against acne vulgaris (acne). The most widely used antibiotics in acne therapy are tetracyclines, macrolides, and lincosamides. Unfortunately, C. acnes bacteria over the past several decades have demonstrated increased resistance to these antibiotics, particularly to clindamycin. The precise knowledge of how antibiotics interact with their clinical target is needed to overcome this problem. Toward this goal, we determined the structure of clindamycin in complex with the ribosome of Cutibacterium acnes at 2.53 Å resolution using cryogenic electron microscopy. The galactose sugar moiety of clindamycin interacts with nucleotides of the 23S rRNA directly or through a conserved network of water-mediated interactions. Its propyl pyrrolidinyl group interacts with the 23S rRNA through van der Waals forces. Clindamycin binding to the Cutibacterium acnes ribosome interferes with both: proper orientation of the aminoacyl group of the A-site bound tRNA that is needed for peptide bond formation and with the extension of the nascent peptide. Our data are important for advancing understanding of antibiotic resistance and development of narrow- spectrum antibacterial drugs, which is an urgent need for contemporary antibiotic stewardship.

Physiological and Transcriptomic Analyses Provide Insights into Nitrite Stress Responses of the Swimming Crab Portunus trituberculatus.

Nitrite is a common environmental pollutant in intensive aquaculture systems. In this study, physiological and transcriptomic analyses were performed to investigate nitrite stress responses in the swimming crab Portunus trituberculatus, an important aquaculture species in China. The results revealed that nitrite can affect neurotransmitter signaling via the expression of neurotransmitter receptors such as octopamine receptor (OAR) and 5-hydroxytryptamine receptor (5-HTR), and depress ecdysteroid signaling by downregulating ecdysteroid receptor (EcR) as well as its downstream transcription factors in hepatopancreas. In addition, nitrite suppressed the expression of hemocyanins, the oxygen-transporting protein, which at least partly contributed to tissue hypoxia, resulting in a switchover of energy metabolism from aerobic to anaerobic pathway. To meet the energy demand, glycogens and lipids were mobilized and transported to the hemolymph, and the catabolism of amino acids and fatty acids was enhanced to provide energy for hepatopancreas. β-oxidation of fatty acids, the major process by which fatty acids are oxidized to generate energy, seems to occur mainly not in mitochondria but in peroxisomes. Although the cellular protective mechanisms, including antioxidant defense, heat shock response (HSR), unfolded protein response (UPR), and autophagy, were activated, nitrite-induced cellular stress overwhelmed the repairing capacity and caused significant increase in the levels of apoptosis. These results indicated that nitrite stress influences neurotransmitter and endocrine signaling, disturbs energy metabolism, damages cellular components, and induces apoptosis in P. trituberculatus. The findings of this study provide new insights into nitrite stress response in the swimming crab and provide valuable information for aquaculture management of this species.

Exploring Aeration Strategies for Enhanced Simultaneous Nitrification and Denitrification in Membrane Aerated Bioreactors: A Computational Approach.

In this study we employ computational methods to investigate the influence of aeration strategies on simultaneous nitrification-denitrification processes. Specifically, we explore the impact of periodic and intermittent aeration on denitrification rates, which typically lag behind nitrification rates under identical environmental conditions. A two-dimensional deterministic multi-scale model is employed to elucidate the fundamental processes governing the behavior of membrane aerated biofilm reactors (MABRs). We aim to identify key factors that promote denitrification under varying aeration strategies. Our findings indicate that the concentration of oxygen during the off phase and the duration of the off interval play crucial roles in controlling denitrification. Complete discontinuation of oxygen is not advisable, as it inhibits the formation of anaerobic heterotrophic bacteria, thereby impeding denitrification. Extending the length of the off interval, however, enhances denitrification. Furthermore, we demonstrate that the initial inoculation of the substratum (membrane in this study) influences substrate degradation under periodic aeration, with implications for both nitrification and denitrification. Comparison between continuous and periodic/intermittent aeration scenarios reveals that the latter can extend the operational cycle of MABRs. This extension is attributed to relatively low biofilm growth rates associated with non-continuous aeration strategies. Consequently, our study provides a comprehensive understanding of the intricate interplay between aeration strategies and simultaneous nitrification-denitrification in MABRs. The insights presented herein can contribute significantly to the optimization of MABR performance in wastewater treatment applications.

The Microbiome in Inflammatory Bowel Disease.

The management of patients with inflammatory bowel disease (IBD) aims to control inflammation through the use of immunosuppressive treatments that target various points in the inflammatory cascade. However, the efficacy of these therapies in the long term is limited, and they often are associated with severe side effects. Although the pathophysiology of the disease is not completely understood, IBD is regarded as a multifactorial disease that occurs due to an inappropriate immune response in genetically susceptible individuals. The gut microbiome is considered one of the main actors in the development of IBD. Gut dysbiosis, characterised by significant changes in the composition and functionality of the gut microbiota, often leads to a reduction in bacterial diversity and anti-inflammatory anaerobic bacteria. At the same time, bacteria with pro-inflammatory potential increase. Although changes in microbiome composition upon biological agent usage have been observed, their role as biomarkers is still unclear. While most studies on IBD focus on the intestinal bacterial population, recent studies have highlighted the importance of other microbial populations, such as viruses and fungi, in gut dysbiosis. In order to modulate the aberrant immune response in patients with IBD, researchers have developed therapies that target different players in the gut microbiome. These innovative approaches hold promise for the future of IBD treatment, although safety concerns are the main limitations, as their effects on humans remain unknown.

A cytochrome bd repressed by a MarR family regulator confers resistance to metals, nitric oxide, sulfide, and cyanide in Chromobacterium violaceum.

The terminal oxidases of bacterial respiratory chains rely on heme-copper (heme-copper oxidases) or heme (cytochrome bd ) to catalyze reduction of molecular oxygen to water. Chromobacterium violaceum is a facultative anaerobic bacterium that uses oxygen and other electron acceptors for respiration under conditions of varying oxygen availability. The C. violaceum genome encodes multiple respiratory terminal oxidases, but their role and regulation remain unexplored. Here, we demonstrate that CioAB, the single cytochrome bd from C. violaceum , protects this bacterium against multiple stressors that are inhibitors of heme-copper oxidases, including nitric oxide, sulfide, and cyanide. CioAB also confers C. violaceum resistance to iron, zinc, and hydrogen peroxide. This cytochrome bd is encoded by the cioRAB operon, which is under direct repression by the MarR-type regulator CioR. In addition, the cioRAB operon responds to quorum sensing and to cyanide, suggesting a protective mechanism of increasing CioAB in the setting of high endogenous cyanide production.

Microbiome of craft hard cheeses from raw goat milk during ripening

The production of craft cheeses from goat milk in small farms in Ukraine is becoming increasingly widespread. The uniqueness of goat cheeses made from raw milk is attributed to the significant diversity of microbiological processes that occur during their ripening, involving natural strains of bacteria, fungi, yeasts, and cheese mites. Therefore, this research aimed to determine the microbial composition of Caciotta and Canestrato goat cheeses during their ripening process. The number of mesophilic aerobic and facultative anaerobic microorganisms (MAFAM) in Caciotta cheese was stable on the 10th day, the 1st, and the 12th month, and decreased by 1.18–1.27 lg CFU/g by the 24th month of ripening. The number of mold fungi and yeasts in Caciotta cheese peaked in the 1st month but they were not detected in the 12th and 24th months of ripening. Lactic acid bacteria in Caciotta cheese formed the basis of MAFAM and were represented by Lactobacillus plantarum on the 10th day and the 1st month of ripening, Lactobacillus brevis and Leuconostoc pseudomesenteroides in the 12th month, and L. brevis and Leuconostoc mesenteroides in the 24th month. In 24-month-ripened Caciotta cheese, Escherichia coli, Enterobacter ludwigii, E. durans, E. faecalis, and E. hirae were detected. Lactic acid bacteria in Canestrato cheese formed the basis of MAFAM and were represented by L. mesenteroides from the 10th day to the 3rd month of age, L. pseudomesenteroides in the 6th month, and L. plantarum in the 12th month. Canestrato cheese was characterized by a significant presence of mold fungi and yeasts in all ripening periods except for the 6th month. In 12-month-ripened Canestrato cheese, Enterobacter cloacae, Bacillus cereus, Enterococcus durans, E. hirae, and E. faecalis were isolated. Cheese mites, Acarus siro, in various stages of development were found in the rind of both cheeses starting from the 6th month of ripening. The results of the researchers provide new data on the microbiome of craft hard cheeses made from raw goat milk and can be used to develop methods for controlling the population size of A. siro mites during their ripening.

Microbiological characterization of neuropathic diabetic foot infection: a retrospective study at a Portuguese tertiary hospital

Diabetic foot infection imposes a significant burden and is the major cause of nontraumatic limb amputation. Adequate patient management with effective antibiotic therapy is crucial.This retrospective cohort study aimed to characterize the microbiology and resistance patterns of moderate to severe neuropathic diabetic foot infection in patients hospitalized at a tertiary referral hospital between January 2020 and June 2023. Deep tissue specimens from ulcers were collected for culture.Sixty inpatients were included (62% male, mean age 59.1 ± 11.5 years). Osteomyelitis was present in 90% of the patients. Among 102 microorganisms (average of 1.91 ± 1.25 pathogens per patient), 60.8% were gram-positive bacteria, 31.4% were gram-negative, 3.92% were anaerobic bacteria, and 3.92% were fungi. Staphylococcus aureus (19%) and Enterococcus faecium (17%) were the most common. Pseudomonas aeruginosa (8%) and bacteria of the Enterobacterales family (24%) accounted for all the isolated gram-negative bacteria. Sixteen percent of Staphylococcus aureus and 67% of coagulase-negative Staphylococci were resistant to methicillin. Resistance to ampicillin was found in 11% of Enterococci. All Pseudomonas aeruginosa isolates were sensitive to piperacillin-tazobactam, ceftazidime, or cefepime. Among the Enterobacterales, resistance rates were 35% for piperacillin-tazobactam, 38% for ceftazidime, 21% for cefepime, and 13% for carbapenems.Although the prevalence of methicillin-resistant staphylococci was lower than that in other studies, carbapenem resistance among gram-negative bacteria warrants attention. This study highlights the importance of understanding local epidemiology for effective diabetic foot infection management and resistance mitigation.

Temporal microbiome changes in axolotl limb regeneration: Stage-specific restructuring of bacterial and fungal communities with a Flavobacterium bloom during blastema proliferation.

The intricate relationship between regeneration and microbiota has recently gained attention, spanning diverse model organisms. Axolotl (Ambystoma mexicanum) is a critically endangered salamander species and a model organism for regenerative and developmental biology. Despite its significance, a noticeable gap exists in understanding the interplay between axolotl regeneration and its microbiome. Here, we analyse in depth bacterial 16S rRNA amplicon dataset that we reported before as data resource and profile fungal community by sequencing ITS amplicons at the critical stages of limb regeneration (0-1-4-7-30-60 days post amputation, 'dpa'). Results reveal a decline in richness and evenness in the course of limb regeneration, with bacterial community richness recovering beyond 30 dpa unlike fungi community. Beta diversity analysis reveals precise restructuring of the bacterial community along the three phases of limb regeneration, contrasting with less congruent changes in the fungal community. Temporal dynamics of the bacterial community highlight prevalent anaerobic bacteria in initiation phase and Flavobacterium bloom in the early phase correlating with limb blastema proliferation. Predicted functional analysis mirrors these shifts, emphasising a transition from amino acid metabolism to lipid metabolism control. Fungal communities shift from Blastomycota to Ascomycota dominance in the late regeneration stage. Our findings provide ecologically relevant insights into stage specific role of microbiome contributions to axolotl limb regeneration.

Guided Plasma Application in Dentistry-An Alternative to Antibiotic Therapy.

Cold atmospheric plasma (CAP) is a promising alternative to antibiotics and chemical substances in dentistry that can reduce the risk of unwanted side effects and bacterial resistance. AmbiJet is a device that can ignite and deliver plasma directly to the site of action for maximum effectiveness. The aim of the study was to investigate its antimicrobial efficacy and the possible development of bacterial resistance. The antimicrobial effect of the plasma was tested under aerobic and anaerobic conditions on bacteria (five aerobic, three anaerobic (Gram +/-)) that are relevant in dentistry. The application times varied from 1 to 7 min. Possible bacterial resistance was evaluated by repeated plasma applications (10 times in 50 days). A possible increase in temperature was measured. Plasma effectively killed 106 seeded aerobic and anaerobic bacteria after an application time of 1 min per 10 mm2. Neither the development of resistance nor an increase in temperature above 40 °C was observed, so patient discomfort can be ruled out. The plasma treatment proved to be effective under anaerobic conditions, so the influence of ROS can be questioned. Our results show that AmbiJet efficiently eliminates pathogenic oral bacteria. Therefore, it can be advocated for clinical therapeutic use.