Development of a human skin commensal microbe for bacteriotherapy of atopic dermatitis and use in a phase 1 randomized clinical trial.

Staphylococcus aureus colonizes patients with atopic dermatitis (AD) and exacerbates disease by promoting inflammation. The present study investigated the safety and mechanisms of action of Staphylococcus hominis A9 (ShA9), a bacterium isolated from healthy human skin, as a topical therapy for AD. ShA9 killed S. aureus on the skin of mice and inhibited expression of a toxin from S. aureus (psmα) that promotes inflammation. A first-in-human, phase 1, double-blinded, randomized 1-week trial of topical ShA9 or vehicle on the forearm skin of 54 adults with S. aureus-positive AD (NCT03151148) met its primary endpoint of safety, and participants receiving ShA9 had fewer adverse events associated with AD. Eczema severity was not significantly different when evaluated in all participants treated with ShA9 but a significant decrease in S. aureus and increased ShA9 DNA were seen and met secondary endpoints. Some S. aureus strains on participants were not directly killed by ShA9, but expression of mRNA for psmα was inhibited in all strains. Improvement in local eczema severity was suggested by post-hoc analysis of participants with S. aureus directly killed by ShA9. These observations demonstrate the safety and potential benefits of bacteriotherapy for AD.

Skin microbiota analysis-inspired development of novel anti-infectives.

BACKGROUND: The alarming spread of antimicrobial resistance requires the development of novel anti-infective drugs. Despite the recent research focus on the human microbiome and its likely value to understand and exploit inter-bacterial inhibitory phenomena as a source for antimicrobial strategies, the human microbiota has barely been investigated for the purpose of drug development. RESULTS: We performed a large screen analyzing over 3000 human skin isolates to evaluate bacterial competition within the human skin microbiota as a basis for the development of anti-infective therapeutics. We discovered a Staphylococcus hominis strain with strong and broad activity against Gram-positive pathogens that was mediated by the bacteriocin micrococcin P1 (MP1). In "probiotic" approaches, this strain led to reduced Staphylococcus aureus infection and accelerated closure of S. aureus-infected wounds. Furthermore, we used a nanoparticle strategy to overcome the physico-chemical limitations often encountered with natural substances such as MP1 and demonstrate a significant reduction of S. aureus infection by MP1-loaded nanoparticles. CONCLUSIONS: Our study gives examples of how analysis of bacterial interactions in the human microbiota can be explored for the development of novel, effective anti-infective strategies. Video Abstract.

Optimization of physical parameters for enhanced production of lipase from Staphylococcus hominis using response surface methodology.

Lipase, a versatile hydrolytic enzyme, is gaining more importance in environmental applications such as treatment of oil and grease containing wastewater, pretreatment of solid waste/industrial wastewater for anaerobic treatment. In the present study, the attempts have been made to improve the production of lipase from Staphylococcus hominis MTCC 8980 by optimization of pH, temperature, and agitation speed in lab scale shake flasks culture. The experiments were designed using the full factorial central composite design of experiment. A total of 20 experiments were conducted, and the optimized pH, temperature, and agitation speed were found to be 7.9, 33.1 °C, and 178.4 rpm, respectively. The results of the analysis of variance (ANOVA) test revealed that the linear terms for temperature and agitation were significant (p value < 0.05). Interaction for pH and agitation speed was found to have a significant effect on lipase production from S. hominis MTCC 8980. A 150% increase in enzyme activity was observed under the optimized conditions with the maximum lipase activity of 1.82 U/ml. Further enhancement of enzyme activity can be expected from the optimization of medium components.

Enhancement of anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array and Box-Behnken design.

The prime focus of the present investigation was to optimize statistically the anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array (OA) and Box-Behnken design (BBD). The anti-tubercular activity of strain MANF2 was determined against Mycobacterium tuberculosis H37Rv using luciferase reporter phase assay. Among varied media examined, the isolate exhibited impressive anti-tubercular activity with paramount relative light unit reduction of >90% in de Man Rogose Sharpe (MRS) broth. Primarily, the anti-tubercular activity and biomass of strain MANF2 were estimated in MRS broth by optimizing eight diversified parameters using one factor at a time (OFAT) method after working out a series of experiments. The most significant contributing factors selected through OFAT tool were optimized using Taguchi approach with a standard OA layout of L18 (22 × 36). Results demonstrated the significant (P ≤ 0.05) influence of pH, temperature, yeast extract, magnesium sulphate, and glycerol on response variables. These controlled variables were further optimized using BBD matrix at N = 46 by second-order polynomial equation. The fermentation medium of pH 6.5 constituting yeast extract (0.5% w/v), magnesium sulphate (0.1% w/v), and glycerol (1.5% v/v), being further incubated at 30 °C showed enhanced anti-tubercular activity (98.7%) and approximately 4 fold increment in the bacterial biomass yield (8.3 mg/mL) with respect to traditional OFAT method. Three-dimensional response plots of the quadratic model showed interdependent interaction between the significant variables. In conclusion, the present study revealed the first report on the optimization of anti-tubercular activity and biomass of S. hominis via Taguchi OA as well as BBD design, and thus, paved a path for its proficient applications in pharmaceutical industries as dynamic mycobactericidal agent in future.

Characterization of biofilm formation, antimicrobial resistance, and staphylococcal cassette chromosome mec analysis of methicillin resistant Staphylococcus hominis from blood cultures of children.

INTRODUCTION:: Methicillin resistant Staphylococcus hominis (MRSHo) has been recognized as an important human pathogen, particularly in immunocompromised patients. METHODS:: A total of 19 S. hominis isolates were collected from children at the Children's Medical Centre, Tehran, Iran, from March 2012 to February 2013. MRSHo susceptibility against 13 antimicrobial and 3 antiseptic agents was determined using disk diffusion (DAD) and minimum inhibitory concentration (MIC), respectively. All isolates were subjected to polymerase chain reaction (PCR) assay for 15 distinct resistance genes, staphylococcal cassette chromosome mec (SCCmec), and arginine catabolic mobile elements (ACMEs). Biofilm production of the isolates was determined using a colorimetric microtiter plate assay. RESULTS:: Of the 19 isolates, 16 were resistant to oxacillin and harbored mecA. High resistance was also observed against trimethoprim/sulfamethoxazole (81.2%). All MRSHo isolates were susceptible to the three disinfectants tested (Septicidine-PC, Septi turbo, and Sayacept-HP). In total, 15 (78.9%) isolates produced biofilms. Three isolates had SCCmec types (V and VIII), 13 were untypable (UT), and 5 had ACME type II. CONCLUSIONS:: The results indicate that MRSHo with high antibiotic resistance and unknown SCCmec might become a serious problem in the future for the treatment of patients such as children.

Characterization of biofilm formation, antimicrobial resistance, and staphylococcal cassette chromosome mec analysis of methicillin resistant Staphylococcus hominis from blood cultures of children

Abstract INTRODUCTION: Methicillin resistant Staphylococcus hominis (MRSHo) has been recognized as an important human pathogen, particularly in immunocompromised patients. METHODS: A total of 19 S. hominis isolates were collected from children at the Children's Medical Centre, Tehran, Iran, from March 2012 to February 2013. MRSHo susceptibility against 13 antimicrobial and 3 antiseptic agents was determined using disk diffusion (DAD) and minimum inhibitory concentration (MIC), respectively. All isolates were subjected to polymerase chain reaction (PCR) assay for 15 distinct resistance genes, staphylococcal cassette chromosome mec (SCCmec), and arginine catabolic mobile elements (ACMEs). Biofilm production of the isolates was determined using a colorimetric microtiter plate assay. RESULTS: Of the 19 isolates, 16 were resistant to oxacillin and harbored mecA. High resistance was also observed against trimethoprim/sulfamethoxazole (81.2%). All MRSHo isolates were susceptible to the three disinfectants tested (Septicidine-PC, Septi turbo, and Sayacept-HP). In total, 15 (78.9%) isolates produced biofilms. Three isolates had SCCmec types (V and VIII), 13 were untypable (UT), and 5 had ACME type II. CONCLUSIONS: The results indicate that MRSHo with high antibiotic resistance and unknown SCCmec might become a serious problem in the future for the treatment of patients such as children.

Evaluation of tetraethoxysilane (TEOS) sol-gel coatings, modified with green synthesized zinc oxide nanoparticles for combating microfouling.

Green synthesis of zinc oxide nanoparticles (ZnO-NPs) is gaining importance as an eco-friendly alternative to conventional methods due to its enormous applications. The present work reports the synthesis of ZnO-NPs using the endosperm of Cocos nucifera (coconut water) and the bio-molecules responsible for nanoparticle formation have been identified. The synthesized nanoparticles were characterized using UV-Visible spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Transmission Electron Microscopy (TEM) and Zeta potential measurement. The results obtained reveal that the synthesized nanoparticles are moderately stable with the size ranging from 20 to 80 nm. The bactericidal effect of the nanoparticles was proved by well diffusion assay and determination of minimum inhibitory concentration (MIC) against marine biofilm forming bacteria. Further the green synthesized ZnO-NPs were doped with TEOS sol-gels (TESGs) in order to assess their antimicrofouling capability. Different volumes of liquid sol-gels were coated on to 96-well microtitre plate and cured under various conditions. The optimum curing conditions were found to be temperature 60 °C, time 72 h and volume 200 µl. Antiadhesion test of the undoped (SG) and ZnO-NP doped TEOS sol-gel (ZNSG) coatings were evaluated using marine biofilm forming bacteria. ZNSG coatings exhibited highest biofilm inhibition (89.2%) represented by lowest OD value against Pseudomonasotitidis strain NV1.

Biofilm formation by Staphylococcus hominis strains isolated from human clinical specimens.

Staphylococcus hominis is the third species of coagulase-negative staphylococci (CoNS) most frequently isolated from specimens of patients with hospital-acquired infections. Many infections caused by CoNS appeared to be associated with biofilms. Nevertheless, the knowledge of the ability of S. hominis to form a biofilm is limited. The aim of this study was to analyze the formation of the biofilm by 56 S. hominis strains isolated from clinical cases. The biofilm three-dimensional structure was reconstructed by confocal laser scanning microscopy. We found that most of S. hominis strains carried icaADBC genes encoding polysaccharide intercellular adhesin (PIA), which plays a crucial role in the formation of biofilms in staphylococci strains. However, only a half of the ica-positive strains had an ability to form a biofilm in vitro. In this study, we also accessed the sensitivity of biofilms of S. hominis strains to sodium metaperiodate, proteinase K and DNase. We found that polysaccharides and proteins are the major components of the extracellular matrix of the biofilm formed by S. hominis. DNase did not have a significant effect on biofilms, which suggested that nucleic acid plays a minor role in the mature biofilm.

Probiotic potential of Staphylococcus hominis MBBL 2-9 as anti-Staphylococcus aureus agent isolated from the vaginal microbiota of a healthy woman.

AIMS: To isolate and characterize an antagonist for use as probiotic agent in the biocontrol of Staphylococcus aureus. METHODS AND RESULTS: Bacteria that exhibited antimicrobial activity against Gram-positive bacteria including Staph. aureus were isolated from 12 healthy women, with Staphylococcus hominis MBBL 2-9 showing the strongest activity. The bacteriocin produced by Staph. hominis MBBL 2-9 was purified by 60% ammonium sulfate saturation, ultrafiltration, HLB cartridge and reverse-phase HPLC. The molecular weight was estimated as 2038.2 Da by MALDI-TOF mass spectrometry. The antagonist survived up to 2 h in artificial gastric juice (pH 2.5) and grew in the presence of 1% porcine bile extract. In addition, Staph. hominis MBBL 2-9 adhered effectively to HT-29 epithelial cell line. CONCLUSION: Staphylococcus hominis MBBL 2-9 exhibited desirable probiotic traits such as acid tolerance, bile resistance and adherence to epithelial cell line. The bacterium also produced a bacteriocin with unique molecular weight and high antimicrobial activity similar to traditional antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report of a bacteriocin-producing Staph. hominis MBBL 2-9 that has potential for use as a probiotic agent against Staph. aureus.

Resistance of bacterial biofilms formed on stainless steel surface to disinfecting agent.

The natural ability of microorganisms for adhesion and biofilm formation on various surfaces is one of the factors causing the inefficiency of a disinfection agent, despite its proven activity in vitro. The aim of the study was to determine the effectiveness of disinfecting substances on bacterial biofilms formed on stainless steel surface. A universally applied disinfecting agent was used in the tests. Bacterial strains: Listeria innocua, Pseudomonas putida, Micrococcus luteus, Staphylococcus hominis strains, were isolated from food contact surfaces, after a cleaning and disinfection process. The disinfecting agent was a commercially available acid specimen based on hydrogen peroxide and peroxyacetic acid, the substance that was designed for food industry usage. Model tests were carried out on biofilm formed on stainless steel (type 304, no 4 finish). Biofilms were recorded by electron scanning microscope. The disinfecting agent in usable concentration, 0.5% and during 10 minutes was ineffective for biofilms. The reduction of cells in biofilms was only 1-2 logarithmic cycles. The use of the agent in higher concentration--1% for 30 minutes caused reduction of cell number by around 5 logarithmic cycles only in the case of one microorganism, M. luteus. For other types: L. innocua, P. putida, S. hominis, the requirements placed on disinfecting agents were not fulfilled. The results of experiments proved that bacterial biofilms are resistant to the disinfectant applied in its operational parameters. Disinfecting effectiveness was achieved after twofold increase of the agent's concentration.

A comparison of adherence by four strains of Staphylococcus intermedius and Staphylococcus hominis to canine corneocytes collected from normal dogs and dogs suffering from atopic dermatitis.

The aim of this study was to compare the adherence of four strains of Staphylococcus intermedius and a single strain of Staphylococcus hominis to corneocytes from both normal dogs and dogs suffering from atopic dermatitis. Cells from the skin surface, corneocytes, were collected from 10 normal dogs and 10 dogs suffering from atopic dermatitis. Four strains of S. intermedius, three isolated from canine pyoderma skin lesions (strains A, B and C), and one isolated form from canine synovial membrane sample from a case of septic arthritis (strain D) were compared. S. hominis, which is not normally associated with canine disease, was also evaluated for its ability to adhere to canine corneocytes. S. hominis did not adhere to canine corneocytes. All four strains of S. intermedius adhered well to canine corneocytes collected from both normal and atopic dogs. All strains of S. intermedius showed statistically greater adherence to corneocytes collected from atopic dogs compared with those collected from normal dogs. It was concluded that the adherence assay employed here showed that S. hominis does not adhere to canine corneocytes, S. intermedius adheres preferentially to atopic corneocytes.