In-depth comparative pathogenome, virulome, and resistome analysis of an extensive drug resistant Ralstonia mannitolilytica strain isolated from blood.

INTRODUCTION: Ralstonia mannitolilytica is an global opportunistic pathogen responsible for various diseases. In this study, we reported the genome of a R. mannitolilytica isolate responsible for bacteremia in an acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: Bacterial identification was performed with a Vitek2™ Automated System and 16S rRNA sequencing with BLASTn against the Non-Redundant Protein Sequence (Nr) database. Genome sequencing and analysis were performed using PacBio RS II sequencer, Hierarchical Genome Assembly Process assembly, as well as multiple annotation databases to better understand the innate features. Antibiotic resistance genes and virulence factors were specifically identified through Antibiotic Resistance Genes database and Virulence Factors of Pathogenic Bacteria databases. RESULTS: The complete genome sequence was assembled into two chromosomes with 3,495,817 bp and 1,342,871 bp in length and GC% of 65.37 % and 66.43 %, respectively. The two chromosomes were fully annotated. In chromosome 1 and 2, 19 and 14 antibiotic resistant genes and 48 and 55 virulence factors were predicted, respectively. Specifically, beta-lactam resistance genes blaOXA-443, blaOXA-444 were acquired. CONCLUSIONS: This study aids in the understanding of the innate features of R. mannitolilytica in AECOPD.

A novel alginate-embedded magnetic biochar-anoxygenic photosynthetic bacteria composite microspheres for multipollutant removal: Mechanisms of photo-bioelectrochemical enhancement and excellent reusability performance.

Existing wastewater treatment technologies face the key challenge of simultaneously removing emerging contaminants and nutrients from wastewater efficiently, with a simplified technological process and minimized operational costs. In this study, a novel alginate-embedded magnetic biochar-anoxygenic photosynthetic bacteria composite microspheres (CA-MBC-PSB microspheres) was prepared for efficient, cost-effective and one-step removal of antibiotics and NH4+-N from wastewater. Our results demonstrated that the CA-MBC-PSB microspheres removed 97.23% of sulfadiazine (SDZ) within 7 h and 91% of NH4+-N within 12 h, which were 21.23% and 38% higher than those achieved by pure calcium alginate-Rhodopseudomonas palustris microspheres (53% and 45.7%), respectively. The enhanced SDZ and NH4+-N removal were attributed to the enhanced photoheterotrophic metabolism and excretion of extracellular photosensitive active substances from R. Palustris through the photo-bioelectrochemical interaction between R. Palustris and magnetic biochar. The long-term pollutants removal performance of the CA-MBC-PSB microspheres was not deteriorated but continuously improved with increasing ruse cycles with a simultaneous removal efficiency of 99% for SDZ and 92% for NH4+-N after three cycles. The excellent stability and reusability were due to the fact that calcium alginate acts as an encapsulating agent preventing the loss and contamination of R. palustris biomass. The CA-MBC-PSB microspheres also exhibited excellent performance for simultaneous removal of SDZ (89% in 7 h) and NH4+-N (90.7% in 12 h) from the secondary effluent of wastewater treatment plant, indicating the stable and efficient performance of CA-MBC-PSB microspheres in practical wastewater treatment.

Invited review: Role of Bacillus licheniformis in the dairy industry-Friends or foes?

Bacillus licheniformis is one of the major spore-forming bacteria with great genotypic diversity in raw milk, dairy ingredients, and final dairy products; it is found throughout the dairy-processing continuum. Although being widely used as a probiotic strain, this species also serves as a potential risk in the dairy industry based on its roles in foodborne illness and dairy spoilage. Biofilm formation of B. licheniformis, combined with the heat resistance of its spores, make it impossible to prevent the presence of B. licheniformis in final dairy products by using traditional cleaning and disinfection procedures. Despite the extensive efforts to identify B. licheniformis in various dairy samples, no reviews have been written on both hazards and benefits of this sporeformer. This review discusses the prevalence of B. licheniformis from raw milk to commercial dairy products, biofilm formation and spoilage potential of B. licheniformis, and possible prevention methods. In addition, the potential benefits of B. licheniformis in the dairy industry are also summarized.

Exploring Enhanced Hydrolytic Dehydrogenation of Ammonia Borane with Porous Graphene-Supported Platinum Catalysts.

Graphene is a good support for immobilizing catalysts, due to its large theoretical specific surface area and high electric conductivity. Solid chemical converted graphene, in a form with multiple layers, decreases the practical specific surface area. Building pores in graphene can increase specific surface area and provide anchor sites for catalysts. In this study, we have prepared porous graphene (PG) via the process of equilibrium precipitation followed by carbothermal reduction of ZnO. During the equilibrium precipitation process, hydrolyzed N,N-dimethylformamide sluggishly generates hydroxyl groups which transform Zn2+ into amorphous ZnO nanodots anchored on reduced graphene oxide. After carbothermal reduction of zinc oxide, micropores are formed in PG. When the Zn2+ feeding amount is 0.12 mmol, the average size of the Pt nanoparticles on PG in the catalyst is 7.25 nm. The resulting Pt/PG exhibited the highest turnover frequency of 511.6 min-1 for ammonia borane hydrolysis, which is 2.43 times that for Pt on graphene without the addition of Zn2+. Therefore, PG treated via equilibrium precipitation and subsequent carbothermal reduction can serve as an effective support for the catalytic hydrolysis of ammonia borane.

Fusion of Quality Evaluation Metrics and Convolutional Neural Network Representations for ROI Filtering in LC-MS.

Region of interest (ROI) extraction is a fundamental step in analyzing metabolomic datasets acquired by liquid chromatography-mass spectrometry (LC-MS). However, noises and backgrounds in LC-MS data often affect the quality of extracted ROIs. Therefore, developing effective ROI evaluation algorithms is necessary to eliminate false positives meanwhile keep the false-negative rate as low as possible. In this study, a deep fused filter of ROIs (dffROI) was proposed to improve the accuracy of ROI extraction by combining the handcrafted evaluation metrics with convolutional neural network (CNN)-learned representations. To evaluate the performance of dffROI, dffROI was compared with peakonly (CNN-learned representation) and five handcrafted metrics on three LC-MS datasets and a gas chromatography-mass spectrometry (GC-MS) dataset. Results show that dffROI can achieve higher accuracy, better true-positive rate, and lower false-positive rate. Its accuracy, true-positive rate, and false-positive rate are 0.9841, 0.9869, and 0.0186 on the test set, respectively. The classification error rate of dffROI (1.59%) is significantly reduced compared with peakonly (2.73%). The model-agnostic feature importance demonstrates the necessity of fusing handcrafted evaluation metrics with the convolutional neural network representations. dffROI is an automatic, robust, and universal method for ROI filtering by virtue of information fusion and end-to-end learning. It is implemented in Python programming language and open-sourced at https://github.com/zhanghailiangcsu/dffROI under BSD License. Furthermore, it has been integrated into the KPIC2 framework previously proposed by our group to facilitate real metabolomic LC-MS dataset analysis.

A variant ECE1 allele contributes to reduced pathogenicity of Candida albicans during vulvovaginal candidiasis.

Vulvovaginal candidiasis (VVC), caused primarily by the human fungal pathogen Candida albicans, results in significant quality-of-life issues for women worldwide. Candidalysin, a toxin derived from a polypeptide (Ece1p) encoded by the ECE1 gene, plays a crucial role in driving immunopathology at the vaginal mucosa. This study aimed to determine if expression and/or processing of Ece1p differs across C. albicans isolates and whether this partly underlies differential pathogenicity observed clinically. Using a targeted sequencing approach, we determined that isolate 529L harbors a similarly expressed, yet distinct Ece1p isoform variant that encodes for a predicted functional candidalysin; this isoform was conserved amongst a collection of clinical isolates. Expression of the ECE1 open reading frame (ORF) from 529L in an SC5314-derived ece1Δ/Δ strain resulted in significantly reduced vaginopathogenicity as compared to an isogenic control expressing a wild-type (WT) ECE1 allele. However, in vitro challenge of vaginal epithelial cells with synthetic candidalysin demonstrated similar toxigenic activity amongst SC5314 and 529L isoforms. Creation of an isogenic panel of chimeric strains harboring swapped Ece1p peptides or HiBiT tags revealed reduced secretion with the ORF from 529L that was associated with reduced virulence. A genetic survey of 78 clinical isolates demonstrated a conserved pattern between Ece1p P2 and P3 sequences, suggesting that substrate specificity around Kex2p-mediated KR cleavage sites involved in protein processing may contribute to differential pathogenicity amongst clinical isolates. Therefore, we present a new mechanism for attenuation of C. albicans virulence at the ECE1 locus.

Polymicrobial interaction between Lactobacillus and Saccharomyces cerevisiae: coexistence-relevant mechanisms.

The coordination of single or multiple microorganisms are required for the manufacture of traditional fermented foods, improving the flavour and nutrition of the food materials. However, both the additional economic benefits and safety concerns have been raised by microbiotas in fermented products. Among the fermented products, Lactobacillus and Saccharomyces cerevisiae are one of the stable microbiotas, suggesting their interaction is mediated by coexistence-relevant mechanisms and prevent to be excluded by other microbial species. Thus, aiming to guide the manufacture of fermented foods, this review will focus on interactions of coexistence-relevant mechanisms between Lactobacillus and S. cerevisiae, including metabolites communications, aggregation, and polymicrobial biofilm. Also, the molecular regulatory network of the coexistence-relevant mechanisms is discussed according to omics researches.

Significant downtrend of antimicrobial resistance rate and rare ß-lactamase genes and plasmid replicons carriage in clinical Pseudomonas aeruginosa in Southern China.

OBJECTIVES: Pseudomonas aeruginosa is a medically important pathogen showing intrinsic low permeability to various antimicrobial agents and its potential to acquire multiple resistance mechanism. A longitudinal surveillance aimed to investigate the antimicrobial resistance and its determinants of Pseudomonas aeruginosa in Southern China. A total of 2163 P. aeruginosa isolates were obtained from patients in Southern China during 2004-2016. METHODS: The antimicrobial susceptibility of the isolates was performed by disk diffusion and Vitek 2 automated system and interpreted according to the Clinical and Laboratory Standard Institute (CLSI) 2015. RESULTS: A significant downtrend of resistant rate (>10.0%) was observed for tested antibiotic agents including ciprofloxacin (>30.0%), gentamicin (29.0%), tobramycin (24.2%) and ceftazidime (24.0%) except for aztreonam and amikacin. A total of 269 randomly selected isolates were further studied on the carriage of ß-lactam resistance genes by using 7 groups of multiplex PCRs targeting on 20 genes. ß-lactam resistance genes were rarely detected with a rate lower than 8%. Among all ß-lactam resistance genes, blaSHV acquired the highest identification rate (18/269, 6.7%), followed by blaOXA-1-like (6/269, 2.2%) and blaPER (6/269, 2.2%). In addition, 8 different plasmid replicons were amplified using 8 groups of multiplex PCRs including 18 sets of primers. Only five plasmid replicons were identified in 5 different P. aeruginosa isolates. Insignificant clonal relatedness among the positive strains identified by regular PCR were further verified by randomly amplified polymorphic DNA (RAPD)-PCR. CONCLUSION: This study has provided comprehensive knowledge on current antimicrobial resistance, ß-lactam resistance genes and plasmid replicons carriage in a large scale of clinical P. aeruginosa isolates.

Emerging resistance mechanisms for 4 types of common anti-MRSA antibiotics in Staphylococcus aureus: A comprehensive review.

Staphylococcus aureus is one of the leading hospital-associated and community-associated pathogens, which has caused a global public health concern. The emergence of methicillin-resistant S. aureus (MRSA) along with the widespread use of different classes of antibiotics has become a significant therapeutic challenge. Antibiotic resistance is a disturbing problem that poses a threat to humans. Treatment options for S. aureus resistant to ß-lactam antibiotics include glycopeptide antibiotic, cyclic lipopeptide antibiotic, cephalosporins and oxazolidinone antibiotic. The most representative types of these antibiotics are vancomycin, daptomycin, ceftaroline and linezolid. The frequent use of the first-line drug vancomycin for MRSA treatment has increased the number of resistant strains, namely vancomycin intermediate resistant S. aureus (VISA) and vancomycin resistant S. aureus (VRSA). A systematic literature review of relevant published studies in PubMed before 2020 was conducted. In recent years, there have been some reports on the relevant resistant mechanisms of vancomycin, daptomycin, ceftaroline and linezolid. In this review, we have summarized the antibiotic molecular modes of action and different gene mutants at the whole-genome level, which will aid in further development on new drugs for effective MRSA treatment based on describing different resistance mechanisms of classic antibiotics.

Impact of pmrA on Cronobacter sakazakii planktonic and biofilm cells: A comprehensive transcriptomic study.

Cronobacter sakazakii is an emerging opportunistic foodborne pathogen causing rare but severe infections in neonates. Furthermore, the formation of biofilm allows C. sakazakii to persist in different environments. We have demonstrated that the mutator phenotype ascribed to deficiency of the pmrA gene results in more biomass in the first 24 h but less during the post maturation stage (7-14 d) compared with BAA 894. The present study aimed to investigate the regulatory mechanism modulating biofilm formation due to pmrA mutation. The transcriptomic analyses of BAA 894 and s-3 were performed by RNA-sequencing on planktonic and biofilm cells collected at different time points. According to the results, when comparing biofilm to planktonic cells, expression of genes encoding outer membrane proteins, lysozyme, etc. were up-regulated, with LysR family transcriptional regulators, periplasmic proteins, etc. down-regulated. During biofilm formation, cellulose synthase operon genes, flagella-related genes, etc. played essential roles in different stages. Remarkably, pmrA varies the expression of a number of genes related to motility, biofilm formation, and antimicrobial resistance, including srfB, virK, mviM encoding virulence factor, flgF, fliN, etc. encoding flagellar assembly, and marA, ramA, etc. encoding AraC family transcriptional regulators in C. sakazakii. This study provides valuable insights into transcriptional regulation of C. sakazakii pmrA mutant during biofilm formation.

Resistome and virulome study on pathogenic Streptococcus agalactiae Guangzhou-SAG036.

Streptococcus agalactiae is considered as a leading case of bacterial infection among neonates. Although relative protection strategies have been performed in many high-income countries, resulting in a massive reduction in the occurrences of early-onset GBS disease, the late-onset disease has not affected. Here, the whole genome of S. agalactiae Guangzhou-SAG036 was sequenced by the Pacific Biosciences Sequel using the P4-C2 chemistry and the continuous long reads were used for de novo assembly using HGAP. Besides, genes prediction and multiply annotation were performed by comparing it with diverse databases. The whole genome has a length of 2,206,504 bp and contains 2162 predicted genes with an average G + C content of 35.85%. Based on the whole genome sequence, 2 large prophages, 20 virulence factors genes, and 8 antibiotic resistant genes were identified. MLST analysis revealed S. agalactiae Guangzhou-SAG036 was identified as ST-17. The virulence factors genes were identified with different functions including adherence, antiphagocytosis, spreading factor, immune evasion, invasion, toxin. Besides, the antibiotic-resistant genes may provide S. agalactiae with resistance to multi-drugs including erythromycin, streptomycin, azithromycin, spiramycin, ampicillin, kanamycin, cationic peptides, and tetracycline. Therefore, the infection of S. agalactiae Guangzhou-SAG036 ST-17 strain maybe caused by the complex virulence factors and multi-drugs resistance. These results contribute to further understand GBS epidemiology and surveillance targets.

Genomic analysis of a hop-resistance Lactobacillus brevis strain responsible for food spoilage and capable of entering into the VBNC state.

BACKGROUND: Lactobacillus brevis is a major contaminant of spoiled beer. And it was able to enter VBNC state and cause false negative detection, which poses a major challenge to the brewing industry. METHODS: The genomic DNA of L. brevis BM-LB13908 was extracted and purified to form a sequencing library that meets the quality requirements and was sequenced. The sequencing results were then screened and assembled to obtain the entire genome sequence of L. brevis. Predicted genes were annotated by GO database, KEGG pathway database and COG functional classification system. RESULTS: The final assembly yielded 275 scaffolds of a total length of 2 840 080 bp with a G + C content of 53.35%. There were 2357, 701, 1519 predicted genes with corresponding GO functional, COG functional, and KEGG biological pathway annotations, respectively. The genome of L. brevis BM-LB13908 contains hop resistance gene horA and multiple genes related to the formation of VBNC state. CONCLUSIONS: This report describes the draft genome sequence of L. brevis BM-LB13908, a spoilage strain isolated from finished beer sample. This study may support further study on L. brevis and other beer spoilage bacteria, and prevent and control beer spoilage caused by microorganisms.

Antibiotic Susceptibility, Biofilm-Forming Ability, and Incidence of Class 1 Integron of Salmonella spp., Escherichia coli, and Staphylococcus aureus Isolated from Various Foods in a School Canteen in China.

In recent years, the food poisoning incidents from school canteens have aroused widespread concern in China. Microbial contamination to the foods is the main factor responsible for these food poisoning events. In this study, identification of microbial pathogens including Salmonella spp., Escherichia coli, and Staphylococcus aureus in samples (frozen pork, fresh pork, fresh chicken, and different fresh vegetables) of a school canteen in China during 2017 to 2018 was performed. The antibiotic susceptibility pattern, class 1 integron, and biofilm formation ability of the isolated pathogens were also investigated. In total, 96 strains were isolated (32 Salmonella spp., 32 E. coli, and 32 S. aureus). The antibiogram study results demonstrated that 61.5% strains were found resistant to at least one type of antibiotics, and 17.7% were resistant to three or more antibiotics. In addition, 31.3% strains possessed class 1 integron. Among the integron-positive isolates, 38.9% Salmonella spp. and 87.5% E. coli contained ∼800 or/and 1500 bp size gene cassette within the integrons. However, four S. aureus strains possessing class 1 integron without gene cassette were found. Although none of the isolated strains were found strong biofilm producer, 44.8% were found to have weak or moderate biofilm formation ability. Despite biofilm formation ability or not, the Salmonella spp. containing positive class 1 integron showed significant resistance to cefazolin and gentamicin. In addition, class 1 integron-positive E. coli isolates having the biofilm formation ability hardly showed sensitive to four antibiotics, such as amikacin, amoxicillin-clavulanate, cefazolin, and gentamicin. Therefore, it is necessary to reduce the prevalence of antibiotic resistance gene cassettes containing antibiotic resistance genes by the prudent use of antibiotics in livestock farms, and the improvement of food processing and storage environment.

Investigation of Formation of Bacterial Biofilm upon Dead Siblings.

Biocides can effectively kill bacteria; however, whether the dead bacterial cells left on the surface influence the later growth of biofilm is unknown. In this study, we have cultured Pseudomonas aeruginosa (PAO1) biofilm on their dead siblings and have investigated their evolution by using magnetic force modulation atomic force microscopy (MF-AFM). The time dependence of the biofilm thickness indicates that the deposited dead siblings can slow down the growth of PAO1 biofilm. The biofilm growing on dead bacteria layers is softer in comparison with those upon alive siblings, as reflected by the static elastic modulus ( E) and dynamic stiffness ( kd) scaled to the disturbing frequency ( f) as kd = kd,0 fγ, where kd,0 is the scaling factor and γ is the power-law exponent. We reveal that the smaller population instead of the variation of extracellular polymeric substances (EPS) within the biofilm upon the dead siblings is responsible for the softer biofilm. The present study provides a better understanding of the biofilm formation, thus, making it significant for designing antimicrobial medical materials and antifouling coatings.

Phenotypic characterization of pathogenic Cronobacter spp. strains.

BACKGROUND: Cronobacter species are Gram-negative opportunistic foodborne pathogens that may cause enterocolitis, bacteremia and meningitis in neonates and premature neonates. Lipopolysaccharide (LPS) serves as the major component of the outer membrane of cell, is a potential virulence factor for Cronobacter. METHODS: Given the potential importance of this molecule in infection and virulence, SDS-PAGE of LPS, MS and TLC characterization of phospholipids and phenotypic characterization of Cronobacter spp. strains were carried out. RESULT: The phospholipids from Cronobacter yielded four major peaks at m/z 719.9, 733.9, 747.9 and 773.9 in the spectrum. All Cronobacter showed O-antigen bands except C. muytjensii ATCC 51329. When Cronobacter defect O-antigen, the outer membrane permeability and cell surface hydrophobicities are increased. All Cronobacter are able to grow under pH 5.0 condition and able to grow under 6% NaCl concentration. C. dublinensis DSM 18705 has a higher infection rate to Caco-2 cells than other Cronobacter. CONCLUSION: Invasion of pathogens into a host cell is critical component to an infectious case. And C. dublinensis DSM 18705 has a higher infection rate to Caco-2 cells than other Cronobacter.

Effect of ultrasonic field on the enzyme activities and ion balance of potential pathogen Saccharomyces cerevisiae.

OBJECTIVES: This study aimed at investigating the enzyme activities and ion concentrations in potential pathogen S.cerevisiae upon ultrasonic treatment. METHODS: The activities of ATPase and antioxidase were identified by ATPase, SOD, and CAT assay kits following the instructions. Extracellular Ca2+ and K+ concentrations were determined in an atomic absorption spectrometer with calcium and potassium hollow-cathode lamps as radiation sources. RESULTS: SOD and CAT activities were enhanced by relatively low ultrasonic power at early time points and reduced to lower levels. Total ATPase, Na+/K+-ATPase, and Ca2+/Mg2+-ATPase activities were reduced by ultrasonic field, with higher reducing rate at stronger ultrasonic power and early time points. In addition, ultrasonic field disturbed the Ca2+ and K+ balances in S.cerevisiae cells. CONCLUSIONS: Ultrasonic field resulted in the reduce even the lost of S.cerevisiae cell viability.

Comparative genomic analyses of two novel qnrVC6 carrying multidrug-resistant Pseudomonas. spp strains.

OBJECTIVES: This study focused on the comparative genomic analyses of two qnrVC6 carrying Pseudomonas spp. strains which might give us insights on the similarity and difference in the genomic contexts of qnrVC6 gene. METHODS: Comparative genomic analyses of the novel qnrVC6 carrying Pseudomonas spp. genomes with emphasis on their antimicrobial resistance genes and virulence factors were performed. RESULTS: Most Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, Clusters of Orthologous Groups of proteins (COG) categories, and (Gene Ontology) GO terms are shared by both genomes. Although qnrVC6 gene is responsible for the increase of quinolone resistance in both strains, but it duplicated in P. putida strain Guangzhou-Ppu420. And the resistance to ß-lactams and aminoglycosides are dependent on different genes. Sharing some adherence, antiphagocytosis, and iron uptake related genes with P. putida strain Guangzhou-Ppu420, P. aeruginosa strain Guangzhou-Pae617 specifically acquires biosurfactant, pigment, protease, regulation, secretion system, and toxin related virulence factors. CONCLUSIONS: Sharing most KEGG pathways, COG categories, and GO terms, P. putida strain Guangzhou-Ppu420 and P. aeruginosa strain Guangzhou-Pae617 differ in antimicrobial resistance genes and virulence factors.

Correlation and in vitro mechanism of bactericidal activity on E. coli with whey protein isolate during ultrasonic treatment.

OBJECTIVE: This study aimed to investigate correlation and in vitro mechanism of bactericidal activity on E. coli with whey protein isolate (WPI) during ultrasonic treatment. METHODS: The structural changes of WPI under ultrasonic field were studied by amino-acid analyzer, circular dichroism, SDS-PAGE, and spectrophotometer. RESULTS: With the increasing of WPI concentration added during ultrasonic treatment, the survival rate of E. coli increased. The influence of WPI on bactericidal activity under ultrasonic treatment might due to the change of tertiary and higher level structures, not by the primary structure, and had little relation with secondary structure. CONCLUSION: The influence of WPI on bactericidal activity during ultrasonic treatment might due to the change of the tertiary structure and higher level structures.

Physical relation and mechanism of ultrasonic bactericidal activity on pathogenic E. coli with WPI.

OBJECTIVE: This study aimed to investigate the physical relation and mechanism of bactericidal activity on pathogenic E. coli by ultrasonic field with whey protein isolate (WPI). METHODS: Ultrasound treatment was performed under the conditions of intensity at 65 W/cm2, pulse duty ratio at 0.5 for 0-15 min with WPI concentration ranged from 0 to 10%. Viscosity, granularity, surface hydrophobicity, free radical scavenging activity, and thermal denaturation were assessed by rotational viscometer, Malvern Mastersizer 2000 particle size analyzer, fluorescent probe ANS method, DPPH method, and differential scanning calorimetry, respectively. RESULTS: The thermal denaturation of WPI was not altered by ultrasound field, but the viscosity of WPI was increased upon 10 min treatment. Additionally, its ability to scavenge free radicals and hydrophobicity were increased. The result also showed that the bacteria viability was improved by WPI during ultrasound treatment. However, the WPI protection was decreased by the prolonged treatment. CONCLUSION: Ultrasound treatment resulted in the increasing of the viscosity, free radicals scavenging activity and hydrophobicity of WPI which led to reduced bactericidal activity on E. coil, while WPI protection was disintegrated by prolonged treatment.