A plant endophyte Staphylococcus hominis strain MBL_AB63 produces a novel lantibiotic, homicorcin and a position one variant.

Here we report a jute endophyte Staphylococcus hominis strain MBL_AB63 isolated from jute seeds which showed promising antimicrobial activity against Staphylococcus aureus SG511 when screening for antimicrobial substances. The whole genome sequence of this strain, annotated using BAGEL4 and antiSMASH 5.0 to predict the gene clusters for antimicrobial substances identified a novel antimicrobial peptide cluster that belongs to the class I lantibiotic group. The predicted lantibiotic (homicorcin) was found to be 82% similar to a reported peptide epicidin 280 having a difference of seven amino acids at several positions of the core peptide. Two distinct peaks obtained at close retention times from a RP-HPLC purified fraction have comparable antimicrobial activities and LC-MS revealed the molecular mass of these peaks to be 3046.5 and 3043.2 Da. The presence of an oxidoreductase (homO) similar to that of epicidin 280- associated eciO or epilancin 15X- associated elxO in the homicorcin gene cluster is predicted to be responsible for the reduction of the first dehydrated residue dehydroalanine (Dha) to 2-hydroxypropionate that causes an increase of 3 Da mass of homicorcin 1. Trypsin digestion of the core peptide and its variant followed by ESI-MS analysis suggests the presence of three ring structures, one in the N-terminal and other two interlocking rings at the C-terminal region that remain undigested. Homicorcin exerts bactericidal activity against susceptible cells by disrupting the integrity of the cytoplasmic membrane through pore formation as observed under FE-SEM.

Metal-Adapted Bacteria Isolated From Wastewaters Produce Biofilms by Expressing Proteinaceous Curli Fimbriae and Cellulose Nanofibers.

Bacterial biofilm plays a pivotal role in bioremediation of heavy metals from wastewaters. In this study, we isolated and identified different biofilm producing bacteria from wastewaters. We also characterized the biofilm matrix [i.e., extracellular polymeric substances (EPS)] produced by different bacteria. Out of 40 isolates from different wastewaters, only 11 (27.5%) isolates (static condition at 28°C) and 9 (22.5%) isolates (agitate and static conditions at 28 and 37°C) produced air-liquid (AL) and solid-air-liquid (SAL) biofilms, respectively, only on salt-optimized broth plus 2% glycerol (SOBG) but not in other media tested. Biomass biofilms and bacteria coupled with AL biofilms were significantly (P ≤ 0.001) varied in these isolates. Escherichia coli (isolate ENSD101 and ENST501), Enterobacter asburiae (ENSD102), Enterobacter ludwigii (ENSH201), Pseudomonas fluorescens (ENSH202 and ENSG304), uncultured Vitreoscilla sp. (ENSG301 and ENSG305), Acinetobacter lwoffii (ENSG302), Klebsiella pneumoniae (ENSG303), and Bacillus thuringiensis (ENSW401) were identified based on 16S rRNA gene sequencing. Scanning electron microscope (SEM) images revealed that biofilm matrix produced by E. asburiae ENSD102, uncultured Vitreoscilla sp. ENSG301, A. lwoffii ENSG302, and K. pneumoniae ENSG303 are highly fibrous, compact, and nicely interlinked as compared to the biofilm developed by E. ludwigii ENSH201 and B. thuringiensis ENSW401. X-ray diffraction (XRD) results indicated that biofilm matrix produced by E. asburiae ENSD102, uncultured Vitreoscilla sp. ENSG301, and A. lwoffii ENSG302 are non-crystalline amorphous nature. Fourier transform infrared (FTIR) spectroscopy showed that proteins and polysaccharides are the main components of the biofilms. Congo red binding results suggested that all these bacteria produced proteinaceous curli fimbriae and cellulose-rich polysaccharide. Production of cellulose was also confirmed by Calcofluor binding- and spectrophotometric assays. E. asburiae ENSD102, Vitreoscilla sp. ENSG301, and A. lwoffii ENSG302 were tested for their abilities to form the biofilms exposure to 0 to 2000 mg/L of copper sulfate (for Cu), zinc sulfate (for Zn), lead nitrate (for Pb), nickel chloride (for Ni), and potassium dichromate (for Cr), several concentrations of these metals activated the biofilm formation. The polysaccharides is known to sequester the heavy metals thus, these bacteria might be applied to remove the heavy metals from wastewater.

A potential resource for bioconversion of domestic wastewater sludge.

Twenty seven filamentous fungal strains representing five genera; Aspergillus, Penicillium, Trichoderma, Myriodontium and Pleurotus were isolated from four sources; domestic wastewater sludge cake (SC) from IWK (Indah Water Konsortium) wastewater treatment plant, palm oil mill effluent compost from Sri Ulu palm Oil Processing Mill, compost of plant debris, and fungal fruiting bodies from a rotten wood stump. Thirty-three strains/isolates were tested for their ability to convert domestic wastewater sludge into compost by assessing biomass production and growth rate on sludge enriched media. The strains/isolates Aspergillus niger, SS-T2008, WW-P1003 and RW-P1 512 produced the highest dry biomass at higher sludge supplemented culture media from their respective group (Aspergillus, Trichoderma, Penicillium and Basidiomycetes, respectively). This implied these strains are better adapted for growth at higher sludge rich substances, and subsequently may be efficient in bioconversion/biodegradation of sludge. The fungi isolated from ecological closely related sources were more amendable to adaptation in a sludge rich culture media.

Treatment of wastewater sludge by liquid state bioconversion process.

This study was conducted to evaluate the effect of an eminent decay fungus, Phanerocheate chrysosporium of organic residues on wastewater sludge for its improvement through decomposition and separation of waste particles by Liquid State Bioconversion (LSB). The effect of fungal treatment was compared to uninoculated (Control) at three different harvests 7, 14 and 21 days after inoculation (DAI). The observed results showed that the weight loss and solid content of wastewater sludge were significantly influenced by Phanerocheate chrysosporium. Both parameters were highly influenced at 7 DAI. The COD and pH of wastewater sludge were also highly influenced by fungal treatment.

Detection and survival of Listeria monocytogenes in milk and dairy products.

Detection of L. monocytogenes in raw and pasteurized milk, Zabady, Karish, Domiati and Romi cheeses were done in this study using direct and cold enrichment methods. Out of 140 samples 3 samples were positive by cold enrichment and they were 2 of raw milk and one of Domiati cheese. Survival of L. monocytogenes was studied during heat treatment of milk by the sealed tube method of inactivation and it was found that L. monocytogenes inactivated completely at 60 degrees C for 15 minutes. There was a statistically inverse correlation between time of storage of dairy products and viable cell count of L. monocytogenes. PH played an important role in survival of that pathogen.

The importance of incubation temperature for detecting beta-lactam-resistant Staphylococcus aureus strains.

Sixty-six isolates of methicillin-resistant Staphylococcus aureus strains originating from Austria, Egypt, the Federal Republic of Germany and Switzerland were tested for their sensitivity to 12 beta-lactam antibiotics at both 30 degrees and 37 degrees C using the agar disk diffusion test. The MICs for six of the beta-lactam antibiotics were determined by the microdilution technique. Resistance to methicillin was always accompanied by resistance to several beta-lactam antibiotics. Resistance to all semi-synthetic penicillins and cephalosporins, including cefotaxime, was higher at 30 degrees C than at 37 degrees C. Cephalothin and cefamandole proved to be the most effective cephalosporin antibiotics against methicillin-resistant S. aureus strains. Newer cephalosporins, i.e. cefotaxime, cefoxitin and cefuroxime, exhibited only a low rate of activity against these strains. The occurrence of phage-type "85" in S. aureus strains from Austria, Egypt and Germany indicates that certain lysotypes are widespread.