Search

As a crucial biomarker for some diseases, the determination of cholesterol in human serum is of great significance for the diagnosis and prevention of these diseases. Hence, a portable cholesterol detection method is necessary for clinical and domestic applications. Here, a portable paper sensor was designed for cholesterol detection by modifying screen-printed electrode (SPE) with nanoporous gold (NPG). To achieve the reliable cholesterol detection, a synergistic strategy was proposed based on the oxidation of cholesterol by cholesterol oxidase (ChOx) and the reduction of oxidation product (H2O2) by NPG. Compared to existing electrochemical sensors, the resulting paper sensor exhibited a wider linear response in a range from 50 μM to 6 mM as well as a higher sensitivity of 32.68 μA mM−1 cm−2 with a lower detection limit of 8.36 μM. Moreover, the portable paper sensor presented strong anti-interference capability and stability in the detection of cholesterol in human serum, and the data detected by the portable paper sensor were consistent with that obtained by an automatic biochemical analyzer. These unique performances confirmed that the proposed paper sensor was a sensitive, reliable, and portable cholesterol detection method, making it a good choice for cholesterol detection.

Filtration is a promising water treatment method to purify drinking water. To develop highly efficient drinking water filter paper, water-resistant cellulose foam paper with a high wet strength property was fabricated using diverse metal oxide (e.g., copper oxide (CuO), zinc oxide (ZnO), and silver oxide (Ag2O)) nanoparticles. These nanoparticles were synthesized using the hydrothermal reaction method. Their morphological structures were studied using a field emission scanning electron microscope (FESEM). The presence of coated nanoparticles on the cellulose foam filter was verified by energy dispersive X-ray spectroscopy (EDX) methods. The antibacterial performance of different types of modified cellulose foam filters was studied against E. coli, P. aeruginosa, B. subtilis, and B. cereus strains using the zone of inhibition test. The antibacterial profile of the cellulose foam filter impregnated with Ag2O nanoparticles, when tested against different types of bacteria, exhibited higher antibacterial activity than the cellulose foam filter impregnated with ZnO and CuO nanoparticles.

In the present work, a facile and green synthesis approach for the production of monodispersed, small-sized (2.9 ± 0.7 nm) and stable silver nanoparticles (AgNPs) using soluble soybean polysaccharide (SSPS) was reported. SSPS was used as the reducing and stabilizing agent. The obtained SSPS-stabilized AgNPs (SA) were characterized by UV–vis absorption spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, energy dispersive X-ray spectroscopy and transmission electron microscopy. The antimicrobial activity of the SA colloidal dispersion (SACD) was evaluated based on the growth kinetics of bacteria E. coli and S. aureus. Afterwards, the colloidal dispersion was applied as a coating material to Kraft paper. The SACD-coated Kraft paper exhibited excellent antimicrobial activity against above bacteria strains and P. aeruginosa. The effects of SACD coating on surface wettability, barrier property and microstructure of the Kraft paper were also studied. The results suggested that the SSPS-stabilized AgNPs have great potential in antibacterial applications.

The exploitation of fibre based Raman probes has been challenged by often complicated fabrication procedures and difficulties in reproducibility. Here, we have demonstrated a simple and cost-effective approach for sensing pH through an optical fibre, by employing a wax patterned filter paper-based substrate for surface enhanced Raman spectroscopy (SERS). Through this method, high reproducibility between fibres was achieved. In addition to sensing pH, it was possible to extract fluid samples containing P. aeruginosa for further analysis. This dual purpose fibre is bronchoscope deployable, and is able to gather information about both the host and pathogen, which may lead to an improved treatment plan in future in vivo applications.

Toward our goal of scalable, antimicrobial materials based on photodynamic inactivation, paper sheets comprised of photosensitizer-conjugated cellulose fibers were prepared using porphyrin and BODIPY photosensitizers, and characterized by spectroscopic (infrared, UV-vis diffuse reflectance, inductively coupled plasma optical emission) and physical (gel permeation chromatography, elemental, and thermal gravimetric analyses) methods. Antibacterial efficacy was evaluated against Staphylococcus aureus (ATCC-2913), vancomycin-resistant Enterococcus faecium (ATCC-2320), Acinetobacter baumannii (ATCC-19606), Pseudomonas aeruginosa (ATCC-9027), and Klebsiella pneumoniae (ATCC-2146). Our best results were achieved with a cationic porphyrin-paper conjugate, Por((+))-paper, with inactivation upon illumination (30 min, 65 ± 5 mW/cm(2), 400-700 nm) of all bacterial strains studied by 99.99+% (4 log units), regardless of taxonomic classification. Por((+))-paper also inactivated dengue-1 virus (99.995%), influenza A (∼ 99.5%), and human adenovirus-5 (∼ 99%). These results demonstrate the potential of cellulose materials to serve as scalable scaffolds for anti-infective or self-sterilizing materials against both bacteria and viruses when employing a photodynamic inactivation mode of action.

Staphylococcus aureus (S. aureus) is one of the most common etiological agents in hospital-acquired infections and food-borne illness. S. aureus toxins and virulence proteases often circulate in host blood vessels leading to life-threatening diseases. Standard identification approaches include bacterial culturing method, which takes several days. Other nucleic acid-based methods were expensive and required trained personnel. To surmount these limitations, a paper-based biosensor was developed. The sensing mechanism was based on the proteolytic activity of S. aureus proteases on a specific peptide substrate, sandwiched between magnetic nanobeads and gold surface on top of a paper support. An external magnet was fixed on the back of the sensor to accelerate the cleavage of the magnetic nanobeads-peptide moieties away from the sensor surface upon test sample dropping. The colour change resulting from the dissociation of the magnetic nanobeads moieties was detected by the naked eye and analysed using ImageJ analysis software for the purpose of quantitative measurement. Experimental results showed detection limits as low as 7, 40 and 100 CFU/mL for S. aureus in pure broth culture, and inoculated in food produces and environmental samples, respectively upon visual observation. The specificity of the sensor was examined by blind testing a panel of food-contaminating pathogens (Listeria monocytogenesis 19115 and E. coli O157:H7), clinical isolates (methicillin-resistant S. aureus (MRSA) and Candida albicans) and standard (Pseudomonas aeruginosa 15692) pathogens. Negative read-out was observed by the naked eye for all tested isolates except for MRSA. Moreover, this sensing tool requires minute's time to obtain the results. In conclusion, this sensing platform is a powerful tool for the detection of S. aureus as a potential point-of-care diagnostic platform in hospitals and for use by regulatory agencies for better control of health-risks associated with contaminated food consumption.

Point-of-care testing (POCT) of infectious bacterial agents offers substantial benefits for disease diagnosis, mainly by shortening the time required to obtain results and by making the test available bedside or at remote care centers. Immunochromatographic lateral flow biosensors offer a low cost, highly sensitive platform for POCT. In this article, we describe the fabrication and testing of a multiplex immuno-disc sensor for the specific detection of Pseudomonas aeruginosa and Staphylococcus aureus. Antibody conjugated gold nanoparticles were used as the signaling agents. The detection range of the bacteria lies within 500-5000 CFU/ml. The advantage of the immuno-disc sensor is that it does not require any preprocessing of biological sample and is capable of whole cell bacterial detection. We also describe the design and fabrication of a compact portable device which converts the color intensity of the gold nanoparticles that accumulate at the test region into a quantitative voltage reading proportional to the bacterial concentration in the sample. The combination of the immuno-disc and the portable color reader provides a rapid, sensitive, low cost, and quantitative tool for the detection of a panel of infectious agents present in the patient sample.

Pyocyanin is a toxin produced by Pseudomonas aeruginosa. Here we describe a novel paper-based electrochemical sensor for pyocyanin detection, manufactured with a simple and inexpensive approach based on electrode printing on paper. The resulting sensors constitute an effective electrochemical method to quantify pyocyanin in bacterial cultures without the conventional time consuming pretreatment of the samples. The electrochemical properties of the paper-based sensors were evaluated by ferri/ferrocyanide as a redox mediator, and showed reliable sensing performance. The paper-based sensors readily allow for the determination of pyocyanin in bacterial cultures with high reproducibility, achieving a limit of detection of 95 nM and a sensitivity of 4.30 μA/μM in standard culture media. Compared to the similar commercial ceramic based sensors, it is a 2.3-fold enhanced performance. The simple in-house fabrication of sensors for pyocyanin quantification allows researchers to understand in vitro adaptation of P. aeruginosa infections via rapid screenings of bacterial cultures that otherwise are expensive and time-consuming.

We present a low-cost, disposable microbial fuel cell fabricated on a paper based platform, having a start-up time of 10 s. The platform deploys ordinary pencil strokes for graphite electrode deposition. The device uses a membrane-less design in a one-time injection (OTI) mode or a continuous capillary driven flow mode (CPF), where oxygen from the atmosphere is used up at the cathode for water formation, leading to the generation of bioelectricity. The performance of the fuel cell is evaluated using two bacterial strains, namely, Pseudomonas aeruginosa IIT BT SS1 and Shewanella putrefaciens. This flexible device is shown to retain bacteria for a period of at least one hour, resulting in the generation of almost 0.4 V using P. aeruginosa and a maximum current of 18 μA using S. putrefaciens without the use of any additional catalysts.

The objective of this study was to analyze the antimicrobial effect of 2% sodium hypochlorite (NaOCl) and 2% chlorhexidine (CHX) by agar diffusion test and by direct exposure test. Five microorganisms: Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans, and one mixture of these were used. These strains were inoculated in brain heart infusion (BHI) and incubated at 37ºC for 24 h. For the agar diffusion test (ADT), 18 Petri plates with 20 ml of BHI agar were inoculated with 0.1 ml of the microbial suspensions, using sterile swabs that were spread on the medium, obtaining growth in junction. Fifty-four paper disks (9 mm in diameter) were immersed in the experimental solutions for 1 min. Subsequently, three papers disks containing one of the substances were placed on the BHI agar surface in each agar plate. The plates were maintained for 1 h at room temperature, and then incubated at 37ºC for 48 h. The diameter of microbial inhibition was measured around the papers disks containing the substances. For the direct exposure test, 162 #50 sterile absorbent paper points were immersed in the experimental suspensions for 5 min, and were then placed on Petri plates and covered with one of the irrigant solutions, or with sterile distilled water (control group). After intervals of 5, 10 and 30 min, the paper points were removed from contact with the solutions and individually immersed in 7 ml of Letheen Broth, followed by incubation at 37ºC for 48 h. Microbial growth was evaluated by turbidity of the culture medium. A 0.1 ml inoculum obtained from the Letheen Broth was transferred to 7 ml of BHI, and incubated at 37ºC for 48 h. Bacterial growth was again evaluated by turbidity of the culture medium. Gram stain of BHI cultures was used for verification of contamination and growth was determined by macroscopic and microscopic examination. The best performance of antimicrobial effectiveness of NaOCl was observed in the direct exposure test, and of CHX was observed in the agar diffusion test. The magnitude of antimicrobial effect was influenced by the experimental methods, biological indicators and exposure time. O objetivo deste estudo foi testar o efeito antimicrobiano do hipoclorito de sódio a 2% e da clorexidina a 2% por meio de dois métodos: difusão em agar e exposição direta. Cinco microrganismos: Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans, e uma mistura foram utilizados. As cepas foram inoculadas em BHI e incubadas a 37ºC por 24 h. Para o teste de difusão em agar, 18 placas de Petri com 20 ml de BHIA foram inoculadas com 0,1 ml das suspensões microbianas, com auxílio de swab esterilizado, de modo a se obter um crescimento confluente. 54 discos de papel com 9 mm de diâmetro foram imersos nas soluções experimentais durante 1 min. A seguir, em cada placa, 3 discos de papel contendo uma das soluções irrigantes foram colocadas sobre a superfície do BHIA. As placas foram mantidas por 1 h em temperatura ambiente, e incubadas a 37ºC por 48 h. Os diâmetros dos halos de inibição microbiana foram medidos sobre os discos de papel contendo as substâncias, valendo-se de duas medidas de forma perpendicular entre si, sendo obtido a média de seus tamanhos. Para o teste de exposição direta, 162 pontas de papel absorvente esterilizadas nº 50 foram imersas na suspensão experimental por 5 min, e foram colocadas sobre uma placa de Petri e cobertas com 10 ml de uma das soluções irrigantes, ou com a água destilada. Em intervalos de 5, 10 e 30 min, as pontas de papel foram removidas do contato com as soluções teste e individualmente transportadas e imersas 7 ml de Letheen Broth, e incubadas a 37ºC for 48 h. O crescimento microbiano foi avaliado pela turbidade do meio de cultura. Um inóculo de 0.1 ml obtido do Letheen Broth foi transferido para 7 ml of BHI, e incubado nas mesmas condições descritas. O crescimento microbiano foi novamente avaliado pela turbidade do meio de cultura. Os resultados mostraram efetividade antimicrobiana para as duas soluções irrigadoras testadas. A magnitude do efeito antimicrobiano foi influenciada pelo método experimental, pelos microrganismos e pelo tempo de exposição.

Phosphate is an essential nutrient for heterotrophic bacteria, affecting bacterioplankton in aquatic ecosystems and bacteria in biofilms. However, the influence of phosphate limitation on bacterial competition and biofilm development in multispecies populations has received limited attention in existing studies. To address this issue, we isolated 13 adhesive bacteria from paper machine aggregates. Intergeneric inhibition of Pseudomonas aeruginosa WW5 by Serratia marcescens WW4 was identified under phosphate-limited conditions, but not in Luria–Bertani medium or M9 minimal medium. The viable numbers of the pure S. marcescens WW4 culture decreased over 3 days in the phosphate-limited medium; however, the mortality of S. marcescens WW4 was significantly reduced when it was co-cultured with P. aeruginosa WW5, which appeared to sustain the S. marcescens WW4 biofilm. In contrast, viable P. aeruginosa WW5 cells immediately declined in the phosphate-limited co-culture. To identify the genetic/inhibitory element(s) involved in this process, we inserted a mini-Tn5 mutant of S. marcescens WW4 that lacked inhibitory effect. The results showed that an endonuclease bacteriocin was involved in this intergeneric inhibition by S. marcescens WW4 under phosphate limitation. In conclusion, this study highlights the importance of nutrient limitation in bacterial interactions and provides a strong candidate gene for future functional characterisation.

Paper is omnipresent on hospital units, but few studies have examined the possible role of paper in the spread of nosocomial pathogens.To determine by laboratory investigation how long bacterial pathogens can survive on office paper and whether bacteria can be transferred from hands to paper and back to hands in a "worst-case scenario."Samples of four bacterial pathogens (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus hirae) were prepared according to standard laboratory procedures. Sterile swatches of office paper were inoculated with the pathogens and bacterial survival was tested over seven days. To test the transmission of bacteria from one person's hands to paper and back to another person's hands, the fingertips of volunteers were inoculated with a nonpathogenic strain of E. coli; these volunteers then pressed the inoculum onto sterile paper swatches. Another group of volunteers whose hands had been moistened pressed their fingertips onto the contaminated paper swatches. Bacteria transferred to the moistened fingertips were cultivated according to standard laboratory procedures.The four tested organisms showed differences in length of survival depending on environmental room conditions, but were stable on paper for up to 72 hours and still cultivable after seven days. Test organisms were transferred to paper, survived on it, and were retransferred back to hands.Paper can serve as a vehicle for cross-contamination of bacterial pathogens in medical settings if current recommendations on hand hygiene aren't meticulously followed.

A finger rinse technique for counting micro-organisms on hands showed no significant difference in the level of recovered micro-organisms following hand drying using either warm air or paper towels. Contact plate results appeared to reflect the degree of dampness of hands after drying rather than the actual numbers of micro-organisms on the hands. In laboratory tests, a reduction in airborne count of Pseudomonas aeruginosa and Staphylococcus aureus of between 40 and 75% was achieved from 600 readings comparing inlets and outlets of warm air hand driers. In washroom trials, the number of airborne micro-organisms was reduced by between 30 and 75%. Air emitted from the outlet of the driers contained significantly fewer micro-organisms than air entering the driers. Drying of hands with hand driers was no more likely to generate airborne micro-organisms than drying with paper towels. Levels of micro-organisms on external surfaces of hand driers were not significantly different to those on other washroom surfaces. This work shows that warm air hand driers, of the type used in this study, are a hygienic method of drying hands and therefore appropriate for use in both the healthcare and food industry.

Three types of paper-covered and one type of plastic-covered Kodak dental film were used to determine if bacteria could penetrate the coverings and contaminate the inner film. Films from each group were immersed for 30 or 120 seconds in high concentrations of known bacterial suspensions with or without 10% sterilized calf serum added to the incubation media. The plastic-covered film effectively excluded all bacteria, whereas the paper-covered film showed contamination with all organisms even at 30 seconds. Increasing the viscosity of the incubation medium with calf serum decreased the level of contamination.

We have experimentally immunized rats intratracheally with sonicated Pseudomonas aeruginosa to develop a high antibody response systemically and locally. The systemic antibodies were measured by a standard enzyme-linked immunosorbent assay (ELISA) on serum samples, whereas the local antibodies were measured on eluates from paper discs containing saliva. High levels of IgA antibodies were elicited in saliva whereas only traces of IgG antibodies were detected; the opposite results were found in serum. The saliva disc method permits serial measurements from the same animal and therefore offers the possibility to follow post-immunization antibody responses.

A number of germ carriers have been tested quantitatively, in order to select the optimum germ carrier for future quantitative tests of chemical disinfectants, belonging to the class of surface and instrument disinfectants. Quality criteria applied were the storage capacity of the carrier and the germ recovery fraction. For the surface disinfectant, the cotton piece was confirmed to be suited best, while for the instrument disinfection the mineralized soft rubber tube proved to be optimal. The cotton piece, however, stores ten times more germs than the rubber tube which means that with the former, reduction factors of as high as log 6 (-7) can be measured.

A cell-free system capable of carrying out amino acid incorporation into peptides was developed from exponentially growing cultures of Pseudomonas aeruginosa . The amino acid incorporation activity required an ATP energy generating system, Mg 2+ , NH 4 + , tRNA and an amino acid mixture for maximum activity. Ribonuclease, puromycin and chloramphenicol inhibited protein synthesis. N -Formylmethionyl-tRNA was synthesized with a P. aeruginosa enzyme system, methionine, P. aeruginosa tRNA, formate and tetrahydrofolic acid. N -formylmethionine was incorporated into peptides by the P. aeruginosa cell-free system. These results indicate that P. aeruginosa extracts contain methionyl-tRNA transformylase and the complete system for the incorporation of N -formylmethionine into peptides.

Because they are cumbersome, tests to determine the quantitative susceptibility of organisms to antimicrobial drugs are not performed routinely in many diagnostic laboratories. This paper describes a simple method of incorporating the antimicrobial drug in agar. It is an adaptation of the Rolinson and Russell technique which allows the determination of minimum inhibitory concentrations (MIC) of antimicrobial drugs for a large number of organisms. Results are comparable with those obtained when the standard agar dilution method is used. Strains of aerobic Gram-negative bacilli were tested by both methods using ampicillin (86 strains), cephaloridine (72 strains), trimethoprim (72 strains), and gentamicin (72 strains). Of the 302 tests thus performed, a difference in MIC of more than one double dilution was noted in only 11 tests. With one strain of Pseudomonas aeruginosa, however, it was not possible to detect ampicillin resistance by the method described in this paper.

Antibodies to Pseudomonas aeruginosa from 10 cystic fibrosis patients with chronic P. aeruginosa lung infections were quantitatively and qualitatively analyzed. The development of specific antibodies in patient serum was evaluated in a longitudinal study (1972 to 1987). The concentrations and specificities of immunoglobulin G (IgG) and IgM antibodies to purified lipopolysaccharides (LPS) from clinical isolates of P. aeruginosa and to a variety of other gram-negative bacteria were studied by immunoblotting and enzyme-linked immunosorbent assay techniques. Results were compared with the number of immunoprecipitates to P. aeruginosa whole-cell extracts detected by crossed immunoelectrophoresis. IgG, but not IgM, anti-Pseudomonas LPS concentrations increased significantly at the onset of chronic infection and continued to increase during the course of the infection. There was a good positive correlation between the concentration of IgG anti-Pseudomonas LPS antibodies and the number of crossed-immunoelectrophoresis precipitins. The increases in IgG anti-LPS antibody concentrations were much higher to Pseudomonas LPS than to other LPSs. Binding studies demonstrated an increase in binding of IgG anti-Pseudomonas LPS during infection, whereas the binding of other anti-LPS antibodies decreased. Immunoblotting studies confirmed that antibodies reacted strongly with Pseudomonas LPS and weakly with Escherichia coli core-lipid A. The specificity of the reaction with Pseudomonas LPS increased with the duration of infection. It is concluded that anti-LPS response in cystic fibrosis patients during chronic P. aeruginosa infection demonstrates a marked increase in IgG anti-Pseudomonas LPS antibody concentration, specificity, and affinity. The anti-LPS enzyme-linked immunosorbent assay is proposed as a routine test to diagnose and to follow the course of chronic P. aeruginosa lung infection in patients with cystic fibrosis.