Use of Caffeine and Human Pharmaceutical Compounds to Identify Sewage Contamination

Fecal coliform bacteria are widely used as indicators of sewage contamination in surface water. However, there are some disadvantages in these microbial techniques including time consuming (18-48h) and inability in discriminating between human and animal fecal material sources. Therefore, it is necessary to seek a more specific indicator of human sanitary waste. In this study, the feasibility was investigated to apply caffeine and human pharmaceutical compounds to identify the human-source contamination. The correlation between caffeine and fecal coliform was also explored. Surface water samples were collected from upstream, middle-stream and downstream points respectively, along Rochor Canal, as well as 8 locations of Marina Bay. Results indicate that caffeine is a suitable chemical tracer in Singapore because of its easy detection (in the range of 0.30-2.0 ng/mL), compared with other chemicals monitored. Relative low concentrations of human pharmaceutical compounds (< 0.07 ng/mL) in Rochor Canal and Marina Bay water samples make them hard to be detected and difficult to be chemical tracer. However, their existence can help to validate sewage contamination. In addition, it was discovered the high correlation exists between caffeine concentration and fecal coliform density in the Rochor Canal water samples, demonstrating that caffeine is highly related to the human-source contamination.

Fractal Analysis of 16S rRNA Gene Sequences in Archaea Thermophiles

A nucleotide sequence can be expressed as a numerical sequence when each nucleotide is assigned its proton number. A resulting gene numerical sequence can be investigated for its fractal dimension in terms of evolution and chemical properties for comparative studies. We have investigated such nucleotide fluctuation in the 16S rRNA gene of archaea thermophiles. The studied archaea thermophiles were archaeoglobus fulgidus, methanothermobacter thermautotrophicus, methanocaldococcus jannaschii, pyrococcus horikoshii, and thermoplasma acidophilum. The studied five archaea-euryarchaeota thermophiles have fractal dimension values ranging from 1.93 to 1.97. Computer simulation shows that random sequences would have an average of about 2 with a standard deviation about 0.015. The fractal dimension was found to correlate (negative correlation) with the thermophile-s optimal growth temperature with R2 value of 0.90 (N =5). The inclusion of two aracheae-crenarchaeota thermophiles reduces the R2 value to 0.66 (N = 7). Further inclusion of two bacterial thermophiles reduces the R2 value to 0.50 (N =9). The fractal dimension is correlated (positive) to the sequence GC content with an R2 value of 0.89 for the five archaea-euryarchaeota thermophiles (and 0.74 for the entire set of N = 9), although computer simulation shows little correlation. The highest correlation (positive) was found to be between the fractal dimension and di-nucleotide Shannon entropy. However Shannon entropy and sequence GC content were observed to correlate with optimal growth temperature having an R2 of 0.8 (negative), and 0.88 (positive), respectively, for the entire set of 9 thermophiles; thus the correlation lacks species specificity. Together with another correlation study of bacterial radiation dosage with RecA repair gene sequence fractal dimension, it is postulated that fractal dimension analysis is a sensitive tool for studying the relationship between genotype and phenotype among closely related sequences.

Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Determination of Alkaline Protease Production In Serratia Marcescens Sp7 Using Agro Wastes As Substrate Medium, Optimization Of Production Parameters And Purification Of The Enzyme

The enzyme alkaline protease production was determined under solid state fermentation using the soil bacteria Serratia marcescens sp7. The maximum production was obtained from wheat bran medium than ground nut shell and chemically defined medium. The physiological fermentation factors such as pH of the medium (pH 8), Temperature (40oC) and incubation time (48 hrs) played a vital role in alkaline protease production in all the above. 100Mm NaCl has given better resolution during elution of the enzymes. The enzyme production was found to be associated with growth of the bacterial culture.

Smart Motion

Austenite and Martensite indicate the phases of solids undergoing phase transformation which we usually associate with materials and not with living organisms. This article provides an overview of bacterial proteins and structures that are undergoing phase transformation and suggests its probable effect on mechanical behavior. The context is mainly within the role of phase transformations occurring in the flagellum of bacteria. The current knowledge of molecular mechanism leading to phase variation in living organisms is reviewed. Since in bacteria, each flagellum is driven by a separate motor, similarity to a Differential drive in case of four-wheeled vehicles is suggested. It also suggests the application of the mechanism in which bacteria changes its direction of movement to facilitate single point turning of a multi-wheeled vehicle. Finally, examples are presented to illustrate that the motion due to phase transformation of flagella in bacteria can start a whole new research on motion mechanisms.

Characterization of Silica Nanoparticles in Interaction with Escherichia coli Bacteria

The objective of the present investigation was to evaluate the morphology of Escherchia coli bacteria in interaction with SiO2 nanoparticles. This study was made by atomic force microscopy and quartz crystal microbalance using SiO2 nanoparticles with 10nm, 50nm and 100nm diameter and bacteria immobilized on polyelectrolyte multilayer films obtained by spin coating or by “layer by layer” (LbL) method.

Molecular Dynamics and Circular Dichroism Studies on Aurein 1.2 and Retro Analog

Aurein 1.2 is a 13-residue amphipathic peptide with antibacterial and anticancer activity. Aurein1.2 and its retro analog were synthesized to study the activity of the peptides in relation to their structure. The antibacterial test result showed the retro-analog is inactive. The secondary structural analysis by CD spectra indicated that both of the peptides at TFE/Water adopt alpha-helical conformation. MD simulation was performed on aurein 1.2 and retro-analog in water and TFE in order to analyse the factors that are involved in the activity difference between retro and the native peptide. The simulation results are discussed and validated in the light of experimental data from the CD experiment. Both of the peptides showed a relatively similar pattern for their hydrophobicity, hydrophilicity, solvent accessible surfaces, and solvent accessible hydrophobic surfaces. However, they showed different in directions of dipole moment of peptides. Also, Our results further indicate that the reversion of the amino acid sequence affects flexibility .The data also showed that factors causing structural rigidity may decrease the activity. Consequently, our finding suggests that in the case of sequence-reversed peptide strategy, one has to pay attention to the role of amino acid sequence order in making flexibility and role of dipole moment direction in peptide activity. KeywordsAntimicrobial peptides, retro, molecular dynamic, circular dichroism.

Adherence of Alveolar Fibroblasts and Microorganisms on Titanium Implants

An implant elicits a biological response in the surrounding tissue which determines the acceptance and long-term function of the implant. Dental implants have become one of the main therapy methods in clinic after teeth lose. A successful implant is in contact with bone and soft tissue represent by fibroblasts. In our study we focused on the interaction between six different chemically and physically modified titanium implants (Tis-MALP, Tis-O, Tis- OA, Tis-OPAAE, Tis-OZ, Tis-OPAE) with alveolar fibroblasts as well as with five type of microorganisms (S. epidermis, S.mutans, S. gordonii, S. intermedius, C.albicans). The analysis of microorganism adhesion was determined by CFU (colony forming unite) and biofilm formation. The presence of α3β1 and vinculin expression on alveolar fibroblasts was demonstrated using phospho specific cell based ELISA (PACE). Alveolar fibroblasts have the highest expression of these proteins on Tis-OPAAE and Tis-OPAE. It corresponds with results from bacterial adhesion and biofilm formation and it was related to the lowest production of collagen I by alveolar fibroblasts on Tis-OPAAE titanium disc.

Adhesion Properties of Bifidobacterium Pseudocatenulatum G4 and Bifidobacterium Longum BB536 on HT-29 Human Epithelium Cell Line at Different Times and pH

Adhesion to the human intestinal cell is considered as one of the main selection criteria of lactic acid bacteria for probiotic use. The adhesion ability of two Bifidobacteriums strains Bifidobacterium longum BB536 and Bifidobacterium psudocatenulatum G4 was done using HT-29 human epithelium cell line as in vitro study. Four different level of pH were used 5.6, 5.7, 6.6, and 6.8 with four different times 15, 30, 60, and 120 min. Adhesion was quantified by counting the adhering bacteria after Gram staining. The adhesion of B. longum BB536 was higher than B. psudocatenulatum G4. Both species showed significant different in the adhesion properties at the factors tested. The highest adhesion for both Bifidobacterium was observed at 120 min and the low adhesion was in 15 min. The findings of this study will contribute to the introduction of new effective probiotic strain for future utilization.

The Efficiency of Irrigation System and Nitrogen Fixation for inoculated Soybeans by using N15 Tracer Techniques

Repeated additions of the unfertilized bacteria led to increase the activity of Nitrogen-fixing bacteria in the root zone with drip irrigation system compared to traditional manual vaccination to increase the proportion of Nitrogen from 29% to 64%, and the efficiency of adding Nitrogen fertilizer did not exceed 9.5% while dropped to 4%, due to the amount of fertilizer added was not exceed 20kg N/h, and the second was the existence of a large amount of available Nitrogen in the soil by fixation, while the efficiency of irrigation system between 2.08 to 2.26 kg/m3.

Process Optimization for Enhanced Production of Cell Biomass and Metabolites of Fluorescent Pseudomonad R81

The fluorescent pseudomonad strain R81 is a root colonizing rhizobacteria which promotes the growth of many plants by various mechanisms. Its broth containing siderophore (ironchelating compound) and 2,4- diacetyl phloroglucinol (DAPG) is used for preparing bioinoculant formulations for agronomical applications. Glycerol was found to be the best carbon source for improved biomass production. Splitting of nitrogen source to NH4Cl and urea had a stabilizing effect on pH during batch cultivation. Ltryptophan at 0.5 % in the medium increased the siderophore production to 850 mg/l. During batch cultivation of the strain in a bioreactor, a maximum of 4 g/l of dry cell mass, 1.8 g/l of siderophore and 20 mg/l of DAPG was achieved when glycerol was 15 g/l and C/N ratio was maintained at 12.5. In case of intermittent feeding of fresh medium during fed-batch cultivation, the dry cell mass was increased to 25 g/l with improved production of DAPG to 70 mg/l.

Modification and Characterization of Bacterial Cellulose Biopolymer as Proton Conducting Membrane

This study describes the preparation of a novel proton conducting membranes based on bacterial cellulose (BC) modified by grafting of 2-acrylamido-2-methyl-1 -propanesulfonic acid (AMPS) through UV-induced graft polymerization. These AMPS-g-BC membranes have been characterized by various techniques including FTIR, SEM and TGA, to find their successful grafting of AMPS on BC, surface morphology and thermal stability, respectively. Physical properties of AMPS-g-BC membranes have been assessed in terms of Lamda value( λ ), ion exchange capacity(IEC) and proton conductivity. The relationship between degree of grafting and AMPS concentration used for grafting has been determined by weight gain method. An optimum proton conductivity equal to 2.89x10-2 S cm-1 and IEC value equal to 1.79 mmol g-1 have been obtained when 20 wt% AMPS concentration is used for grafting (i.e. the corresponding membrane is notated as AMPS20-g-BC).

Inhibitory Effect of Lactic Acid and Nisin on Bacterial Spoilage of Chilled Shrimp

Lactic acid alone and its combined application with nisin were evaluated for reducing population of naturally occurring microorganisms on chilled shrimp. Fresh shrimps were dipped in 0, 1.0% and 2.0% (v/v) lactic acid alone and their combined application with 0.04 (g/L/kg) nisin solution for 10 min. Total plate counts of aerobic bacteria (TPCs), Psychrotrophic counts, population of Pseudomonas spp., H2S producing bacteria and Lactic acid bacteria (LAB) on shrimps were determined during storage at 4 °C. The results indicated that total plate counts were 2.91 and 2.63 log CFU/g higher on untreated shrimps after 7 and 14 days of storage, respectively, than on shrimps treated with 2.0% lactic acid combined with 0.04 (g/L/kg) nisin. Both concentrations of lactic acid indicated significant reduction on Pseudomonas counts during storage, while 2.0% lactic acid combined with nisin indicated the highest reduction. In addition, H2S producing bacteria were more sensitive to high concentration of lactic acid combined with nisin during storage.

Implementation of Technology Concept for the Reduction of Cyanobacteria in Laboratory

Following the research in the Department of environmental engineering in Faculty of mechanical engineering on Technical University of Kosice and experiences with electrocoagulation style of disposal waste water, there were designed and partly examining the equipment of two stage revitalization on the standing and little fusible water of tenet electrolysis on the little tarns. With the cooperation with vet experts was that manners prove and it is innocuous for animals, during which time cyanobacteria are totally paralyzed. For the implementation of science and research results have been obtained by means EU funds for structural development.

Analysis of Plasmids and Restriction Fragment Length Polymorphisms of Acinetobacter baumannii Isolated from Hospitals- AL Jouf Region- KSA

Abstract–The objectives of the current study are to determine the prevalence, etiological agents, drug susceptibility pattern and plasmid profile of Acinetobacter baumannii isolates from Hospital-Acquired Infections (HAI) at Community Hospital, Al Jouf Province, Saudi Arabia. A total of 1890 patients had developed infection during hospital admission and were included in the study. Among those who developed nosocomial infections, 15(9.4), 10(2.7) and 118 (12.7) had respiratory tract infection (RTI), blood stream infections (BSI) and urinary tract (UTI) respectively. A total of 268 bacterial isolates were isolated from nosocomial infection. S. aureus was reported in 23.5% for of the total isolates followed by Klebsiella pneumoniae (17.5%), E. coli (17.2%), P. aeruginosa (11.9%), coagulase negative staphylococcus (9%), A. baumannii (7.1%), Enterobacter spp. (3.4%), Citrobacter freundii (3%), Proteus mirabilis (2.6%), and Proteus vulgaris and Enterococcous faecalis (0.7%). Isolated organisms are multi-drug resistant, predominantly Gram-positive pathogens with a high incidence of methicillin-resistant S. aureus, extended spectrum beta lactamase and vancomycin resistant enterococci organisms. The RFLP (Fragment Length Polymorphisms) patterns of plasmid preparations from isolated A. baumannii isolates had altered RFLP patterns, possibly due to the presence of plasmid(s). Five A. baumannii isolates harbored plasmids all of which were not less than 2.71kbp in molecular weight. Hence, it showed that the gene coding for the isolates were located on the plasmid DNA while the remaining isolates which have no plasmid might showed gene coding for antibiotic resistance being located on chromosomal DNA. Nosocomial infections represent a current problem in Community Hospital, Al Jouf Province, Saudi Arabia. Problems associated with SSI include infection with multidrug resistant pathogens which are difficult to treat and are associated with increased mortality.

A Review on Application of Chitosan as a Natural Antimicrobial

In recent years application of natural antimicrobials instead of conventional ones, due to their hazardous effects on health, has got serious attentions. On the basis of the results of different studies, chitosan, a natural bio-degradable and non-toxic biopolysaccharide derived from chitin, has potential to be used as a natural antimicrobial. Chitosan has exhibited high antimicrobial activity against a wide variety of pathogenic and spoilage microorganisms, including fungi, and Gram-positive and Gramnegative bacteria. The antimicrobial action is influenced by intrinsic factors such as the type of chitosan, the degree of chitosan polymerization and extrinsic factors such as the microbial organism, the environmental conditions and presence of the other components. The use of chitosan in food systems should be based on sufficient knowledge of the complex mechanisms of its antimicrobial mode of action. In this article we review a number of studies on the investigation of chitosan antimicrobial properties and application of them in culture and food mediums.

Synthesis of Silver Nanoparticles by Chemical Reduction Method and Their Antibacterial Activity

Silver nanoparticles were prepared by chemical reduction method. Silver nitrate was taken as the metal precursor and hydrazine hydrate as a reducing agent. The formation of the silver nanoparticles was monitored using UV-Vis absorption spectroscopy. The UV-Vis spectroscopy revealed the formation of silver nanopart├¡cles by exhibing the typical surface plasmon absorption maxima at 418-420 nm from the UV–Vis spectrum. Comparison of theoretical (Mie light scattering theory) and experimental results showed that diameter of silver nanoparticles in colloidal solution is about 60 nm. We have used energy-dispersive spectroscopy (EDX), X-ray diffraction (XRD), transmission electron microscopy (TEM) and, UV–Vis spectroscopy to characterize the nanoparticles obtained. The energy-dispersive spectroscopy (EDX) of the nanoparticles dispersion confirmed the presence of elemental silver signal no peaks of other impurity were detected. The average size and morphology of silver nanoparticles were determined by transmission electron microscopy (TEM). TEM photographs indicate that the nanopowders consist of well dispersed agglomerates of grains with a narrow size distribution (40 and 60 nm), whereas the radius of the individual particles are between 10 and 20 nm. The synthesized nanoparticles have been structurally characterized by X-ray diffraction and transmission high-energy electron diffraction (HEED). The peaks in the XRD pattern are in good agreement with the standard values of the face-centered-cubic form of metallic silver (ICCD-JCPDS card no. 4-0787) and no peaks of other impurity crystalline phases were detected. Additionally, the antibacterial activity of the nanopart├¡culas dispersion was measured by Kirby-Bauer method. The nanoparticles of silver showed high antimicrobial and bactericidal activity against gram positive bacteria such as Escherichia Coli, Pseudimonas aureginosa and staphylococcus aureus which is a highly methicillin resistant strain.

Assessment of Photodynamic Therapy for Staphylococcus Aureus Infected Wounds using Diffuse Reflectance Spectrometry

In this paper we evaluated the efficacy of photodynamic treatment of infected wounds on pig animal model by diffuse reflectance spectrometry. The study was conducted on fifteen wounds contaminated with Staphylococcus aureus bacteria that were incubated for 30 min with methylene blue solution (c = 3.3 x 10-3 M) and exposed to laser radiations (λ = 670 nm, P = 15 mW) for 15 min. The efficiency of photodynamic inactivation of bacteria was evaluated by microbiological exams and diffuse reflectance spectrometry. The results of the microbiological exams showed that the bacterial concentration has decreased from 6.93±0.138 logCFU/ml to 3.12±0.108 logCFU/ml. The spectral examination showed that the diffuse reflectance of wounds contaminated with Staphylococcus aureus has decreased from 5.06±0.036 % to 3.36±0.025 %. In conclusion, photodynamic therapy is an effective method for the treatment of infected wounds and there is a correlation between the CFU count and diffuse reflectance.

Novel Inhibitor of E. coli DNA Adenine Methyltransferase (Ecodam)

EcoDam is an adenine-N6 DNA methyltransferase that methylates the GATC sites in the Escherichia coli genome. DNA-adenine methylation is not present in higher eukaryotes including humans. These observations raise the possibility that dam inhibitors may be used as anti-microbial agents. Polyphosphate (Poly(P)) is an important metabolite and signaling molecule in prokaryotes and eukaryotes. Here, by using gel retardation experiments to investigate the competition of DNA binding by EcoDam in the presence of polyphosphate, we found that Poly (P) strongly interferes with DNA binding by EcoDam, while same concentration of monophosphate does not. In addition, we demonstrated that Poly (P) binding inhibits the activity of EcoDam and our results suggest that Poly (P) led to strong inhibition of the EcoDam catalytic activity, while monophosphate had only moderate effect.

Antibacterial Capacity of Plumeria alba Petals

Antibacterial activity of Plumeria alba (Frangipani) petals methanolic extracts were evaluated against Escherichia coli, Proteus vulgaris,Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus saprophyticus, Enterococcus faecalis and Serratia marcescens by using disk diffusion method. Concentration extracts (80 %) showed the highest inhibition zone towards Escherichia coli (14.3 mm). Frangipani extract also showed high antibacterial activity against Staphylococcus saprophyticus, Proteus vulgaris and Serratia marcescens, but not more than the zones of the positive control used. Comparison between two broad specrum antibiotics to frangipani extracts showed that the 80 % concentration extracts produce the same zone of inhibition as Streptomycin. Frangipani extracts showed no bacterial activity towards Klebsiella pneumoniae, Pseudomonas aeruginosa and Enterococcus faecalis. There are differences in the sensitivity of different bacteria to frangipani extracts, suggesting that frangipani-s potency varies between these bacteria. The present results indicate that frangipani showed significant antibacterial activity especially to Escherichia coli.