Titer Plate Research Articles

Antimicrobial and antibiofilm potential of Curcuma longa Linn. Rhizome extract against biofilm producing Staphylococcus aureus and Pseudomonas aeruginosa isolates.

More than 65% of all human bacterial infection are associated with biofilm. Bacteria in such biofilms are 10 to 1000-fold more resistant to antibiotics than free living bacteria cells. Organisms such as S. aureus and P. aeruginosa are responsible for a significant number of biofilm related infections. In this study, we investigated the antimicrobial and anti-biofilm activity of C. longa L. rhizome extract against biofilm producing S. aureus and P. aeruginosa isolates. The results of MIC and MBC demonstrated promising antibacterial activity of the rhizome extract. TLC and column chromatography detected various curcuminoids while phytochemical analysis also reveals presence of number of bioactive compounds such as alkaloids, flavonoids, phenolics, terpenoids, etc. Micro titer plate assay indicated significant inhibition of biofilm formation in clinical isolates treated with turmeric extract. Thus, on basis of our results turmeric extracts can be considered as natural antibiofilm and antibacterial agent.

Growth kinetics of a Vero cells adapted Bangladeshi strain of peste des petits ruminants (PPR) virus in cell culture.

Growth kinetics of a Vero cells adapted Bangladeshi strain of peste des petits ruminants virus was studied in Vero cells to determine maximum virus yield. One-step growth curve was formulated after determining virus in both supernatant (CFV) and cell lysate (CAV) at different time categories by microtitre plate titration in Vero cells and the viral presence was confirmed by real-time RT-PCR. The virus was first detected in both the supernatants and cell pellets at 12 hpi. The virus titre reached its plateau at 72 hpi. Maximum virus titre of CAV was 6.2 log10 TCID50/ml and that of CFV was 5.2 log10 TCID50/ml at 72 hpi. After that, the titer gradually declined, but maintained at 4.5 log10 TCID50/ml in case of CAV and 4.2 log10 TCID50/ml in case of CFV at 96 hpi. It was concluded that the optimum time point for harvesting Vero cell culture is 72 hpi.

Study ndvB gene expression in Pseudomonas aeruginosa Producing Biofilm

Pseudomonas aeruginosa have ability to form biofilm, this biofilm responsible for wide infections andantibiotic resistance development. A novel mechanism to antibiotic resistance mechanisms utilized bybiofilm represented by the presence of ndvB geneencodes a glucosyltransferase enzyme involved in theformation of this glucans, thisglucan mediated sequestration antibiotic away from their cellular targets.In this study was collected 145 samples from burns patients and identification by biochwmicaltsets andVitek-2 compact. The studied the biofilm formation was measured using micro titer plate method we foundthat 49 isolates (100%) of P.aeruginosa were biofilm positive and measured minimal biofilm inhibitoryconcentration(MBIC) of biofilm P.aeruginosa positive biofilmused cefepime as antibiotic was tested usingmicrotiter plate microdilution method (MBIC) values (1gm/ml).Molecular detection of ndvB gene wasmeasured using PCR 100% of isolates were found to contain ndv B gene. Expression of ndvB gene wassignificantly high in biofilm isolates using sub- MBIC values of cefepime. Expression of ndvB gene wassignificantly high in biofilm isolates using glucose with concentration 1% and 2%. This study we concludedbiofilm formation is an important trait of P.aeruginosa that isa cause of antibiotic resistance. ndvB geneexpression responsible for the biofilm resistance mechanism in P. aeruginosa biofilms.

Optimization of biosurfactant production by marine Streptomyces youssoufiensis SNSAA03: A comparative study of RSM and ANN approach

The aim of the present study was to optimize media components to increase the production of biosurfactant (Bs) by Streptomyces youssoufiensis SNSAA03 employing response surface methodology (RSM) and artificial neural network (ANN). The RSM showed the following as the optimal condition for the production of Bs: glucose 15 g L–1, casein 0.3 g L–1, NaCl 2.25 g L–1 and pH 7. The superiority of the ANN model was proved with more accurate and consistent R2 values 0.998 when compared to RSM R2 value 0.980. The Bs produced by SNSAA03 was identified as glycolipid with 4–methoxy–2, 5–dimethylfuran–3(2H)–one and 1–Nanodecene acid. The micro titre plate assay showed that glycolipid at the concentration of 125 µg mL−1 was effective in controlling the biofilm formation by S. aureus. Thus, the glycolipid produced from SNSAA03 can be used as a potent antibiofilm agent.

Evaluation of Antioxidant, Antibiofilm, Cytotoxic and AntimicrobialActivities of Calligonum Comosum

Objective: Calligonum Comosum is a native omani medicinal plants used traditionally by the locals to treat inflammation, toothache, gum sores and ulcer. As antimicrobial resistance poses a serious threat on global scale by rendering many currently available antibiotics ineffective. This demands the need to look for novel therapeutic agents and there has been an expanded interest in natural products as sources of pharmacologically active principles. The present study was thus initiated to corroborate the therapeutic uses of this medicinal species by evaluating, the phyto-constituents, antibacterial, anti-biofilm and antioxidant activities of C. comosum. Methods: Quantitative and qualitative phytochemical analysis was done, using standard protocols. The antioxidant activity was assessed applying DPPH free radical scavenging assay, hydrogen peroxide radical scavenging assay and the total antioxidant capacity. Biofilm inhibition activity was evaluated using micro titer plate assay and well diffusion method was employed to determine the antibacterial activity. Cytotoxity was assessed in terms of LC50 value using brine shrimp lethality assay. Results: The methanolic extracts of C. comosum showed significant antibacterial activity against all the five tested bacterial strains, with MIC values for all observed to be 1.25 mg/ml except for E. coli. Substantial reduction of the biofilm formation was observed for all bacterial species treated with C. comosum leaf and stem extracts and the extracts also displayed significant cytotoxicity against brine shrimp nauplii, exhibiting LC50 value of 56.797 μg/ml. The total phenolic content and flavonoid content was observed to be 56.6 ± 1.66 mg GAE/g and 49.33 ± 1.34 mg of QE/g of dry extract respectively. C. comosum exhibited potent DPPH scavenging activity, with IC50 value of 44.90 μg/ml and the total antioxidant capacity was 130 ± 2.04 mg AAE/g. Conclusion: The result validates its ethno-medicinal use and suggests that the C. comosum can be exploited for its antibiofilm, antioxidant and antibacterial properties.

Http://www.plantarchives.org/SPECIAL%20ISSUE%2021-1/429%20(2636-2640).pdf

In the present study, Fifty random samples of each small-scale (plain yoghurt and kariesh cheese) were collected randomly from different dairy shops and markets in Zagazig city for isolation and identification of Escherichia coli which is considered a good indicator of faecal contamination and a major cause of food poisoning. Identification was done microscopically and biochemically by different biochemical tests (IMVIC).The incidence of E. coli in yoghurt and kariesh cheese samples were 36% and 50%, respectively. Also, the serological identification of E. coli isolates revealed that O26 is the most predominant serogroup by percentagesof27.77% and 28% in both yoghurt and kariesh cheese samples, respectively. Because of its rising resistance to antibiotics, E.coli represents a real threat to human health. Antimicrobial susceptibility testing (AST) was done by disc diffusion method against 10 antimicrobials and the results revealed that E. coli isolates were highly resistant to amoxicillin-clavulanate, ampicillin, Cefotaxime and Ceftazidime, and were highly sensitive to chloramphenicol, Ciprofloxacin and tetracycline. In addition, 83.72%of E. coli isolates showed multi drug resistance (MDR).Bacterial adhesion to food surfaces and the formation of biofilmisa source of food contamination that has an impact on food safety and industry. Micro titer plate assay was used for testing biofilm formation and revealed that 69.77% of E. coli isolates were non-biofilm producers, 9.30% were weak biofilm producers, 20.93%were moderate biofilm producers and none of isolates was strong biofilm producers. Virulence genes of E. coli isolates were identified and characterized by a multiplex PCR assay. The results showed that among the target genes, stx1was most frequently detected by a percentage of81.82%

Evaluation of anticancer activity of Melaleuka Alternifolia. (i. e. tea tree oil) on Breast cancer cell line (MDA MB)- An in-vitro study

Introduction and Aims: According to Globocan 2012, India along with United States and China collectively accounts for almost one third of the global breast cancer burden. India is facing challenging situation due to 11.54% increases in incidence and 13.82% increase in mortality due to breast cancer during 2008–2012. The current treatment modality has issues of drug resistance and side effects. Many anticancer drugs currently used clinically have been isolated from plant species hence, investigation of plant species as a source of experimental therapeutic agents, in treating cancer is currently gaining a lot of importance. One such naturally available plant extract Melaleuka Alternifolia (TTO) which belongs to the family of essential oils is a very good antibacterial, antifungal, antiviral, antiprotozoal and anti-inflammatory agent. But currently there is a lot of importance has been given for its anticancer effect. Hence our aim is to evaluate anticancer activity of Melaleuka Alternifolia on Breast cancer cell line (MDA MB) by MTT assay an in vitro method. Material and Methods: Before the start of the study ethical clearance was obtained from Institutional Review Board. The cytotoxicity checked for Breast cancer cell line (MDA MB) and NIH 3T3 mouse fibroblasts which were used as a control in our current study. These cell lines were procured from NCCS Pune, India. 1. MTT solution preparation (stock solution): 5 mg in 1 ml of PBS. 2. Cell culture : The cell lines were maintained in 96 wells micro titer plate containing MEM media supplemented with 10% heat inactivated fetal calf serum (FCS), containing 5% of mixture of Gentamicin (10ug), Penicillin ( 100 Units/ ml) and Streptomycin (100µg/ml) in presence of 5% CO2 at 37oC for 48-72 hours. 3. Cytotoxicity Assay: In vitro growth inhibition effect of test compound was assessed by calorimetric or spectrophotometric determination of conversion of MTT into Formazan blue by living cells. Results: The results represent the mean of five readings. The IC50 value of tea tree oil for breast cancer cell line (MDA MB) after 48 hrs was 25µg/ml. Spearmans rho’s Correlation showed P value Conclusion: TTO has a promising anticancer property against Breast cancer cell line (MDA MB) with its IC50 value 25µg/ml. Hence this TTO with its greater efficacy related to its anticancer activity can be brought to the level of clinical trials in the coming future. Keywords: Cytotoxicity, MTT assay, MDAMB cell line, Tea tree oil.

Evaluate the efficacy of Probiotic Lactobacilli on growth and biofilm formation in Streptococcus mutans isolated from gingivitis

The present study aimed to assess the antibacterial effects of Probiotic Lactobacilli against Streptococcus mutans isolates. A total of 13/52 (25%) S. mutans isolates were collected from patients suffering from gingivitis, which were identified using morphological characteristics, biochemical tests and PCR technique. Micro titer plate method was used to detect the biofilm formation in tested bacterial isolates by measuring optical density (OD) using ELISA reader at a wavelength 640 nm. The results revealed that only 10/13 (77%) isolates produced the biofilm. Based on the results obtained in this study, the isolates were classified according to their ability to the biofilm formation as follows: the isolates with thick biofilm (OD ≥ 0.300 nm), the isolates with moderate biofilm (OD 0.2-0.3 nm), the isolates with thin biofilm (OD 0.1-0.2 nm) and Non-productive isolates of biofilm (OD < 0.1 nm). On the other hand, the agar well diffusion method was used to evaluate the effectiveness of Probiotic Lactobacilli against the growth of bacterial isolates by measuring the inhibition zones around the wells containing of probiotics (50, 100 mg/ml and stock). The results indicate that inhibition zones of the tested isolates were weak at 50 mg/ml, moderate at 100 mg/ml and strong at stock solution. A modified crystal violet test in well micro-titer plates was used to detect the ability of isolates to the biofilm formation after treatment with various concentrations of Probiotic Lactobacilli by measuring optical density (OD). The results indicate a decrease in the ability of the tested isolates to the biofilm formation compared to control where the isolates treated with probiotics at 50 mg/ml formed a strong biofilm (OD 0.300 - 0.341 nm) and the isolates treated with probiotics at 100 mg/ml formed moderate biofilm (OD 0.209 - 0.263 nm), while the isolates treated with the stock solution of probiotics formed a thin biofilm (OD 0.144 - 0.198 nm). In conclusion, the results of the current study indicate that the Probiotic Lactobacilli have inhibitory effect on growth and biofilm formation in Streptococcus mutans isolates and the antibacterial activity of Probiotic Lactobacilli is increased with increasing concentration.

Efficacy of novel phages for control of multi-drug resistant Escherichia coli O177 on artificially contaminated beef and their potential to disrupt biofilm formation

Contaminated beef is a prominent source of foodborne pathogens such as Escherichia coli O177. Susceptibility of nine multi-drug resistant E. coli O177 strains against eight individual phages and six phage cocktails was assessed using polystyrene microplate titer plate. Further, 180 beef samples were independently inoculated with E. coli O177 cells in triplicates and treated with eight individual phages and six phage cocktails to determine their efficacy in inhibiting bacteria growth at 4 °C over a 7-day incubation period. Results revealed that all E. coli O177 strains were susceptible to the phages. A significant log reduction in viable E. coli O177 cell counts was observed on beef samples upon phage treatment over the 7-day incubation period. Two individual phages and three phage cocktails reduced E. coli cell counts to levels below the detection limit (1.0 log10 CFU/g). Log reduction of viable E. coli cell counts ranged from 2.10 to 7.81 CFU/g for individual phages and from 2.86 to 7.81 CFU/g for cocktails. Individual phages and phage cocktails inhibited E. coli O177 biofilm formation with phage cocktails showing high efficacy. Furthermore, phage cocktails showed greater efficacy in destroying pre-formed biofilm than individual phages. Based on these findings, we concluded that phage cocktails developed in this study could be used to reduce E. coli O177 contamination and extend the shelf-life of stored raw beef.

Valoración de hormonas esteroides en heces de una pareja de lobo mexicano (Canis lupus baileyi) en cautiverio

El lobo gris mexicano (Canis lupus baileyi) es la subespecie genéticamente más distintiva de los lobos que habitan Norteamérica y que ha sido eliminada de las zonas en que residía originalmente en nuestro país.Actualmente no hay evidencia de poblaciones en vida libre, por lo que todos los individuos existentes están en zoológicos o encierros construidos especialmente para su reproducción. Referente a su comportamiento yestructura social hay algunos estudios pero son menos los relacionados a su fisiología reproductiva. El presente trabajo describe en una pareja (macho-hembra) de lobo mexicano los perfiles de hormonas esteroides sexuales(HES) durante las estaciones de invierno y primavera que son cuando se reproduce. La cuantificación de las HES (progesterona, P4; testosterona, T, y estradiol, E2) se hicieron por inmunoanálisis a partir de heces fecales.Realizándose, además, observaciones de su conducta sexual usando el método focal con registros de puntos. Al ser las muestras de sexo desconocido se hizo su sexado a partir de la concentración diferencial de T. Validado lo anterior, el perfil hormonal de P4 y T en el macho resultó ser de tipo cíclico, mientras que el de E2 no tuvo cambios significativos durante toda la temporada; siendo su concentración inferior a las de P4 y T. En las hembras, la P4 tuvo un patrón cíclico con evidentes incrementos, así como el de T pero siendo sus concentraciones inferiores. El E2 tuvo incrementos considerables, sin embargo, sus concentraciones fueron menores que las otras dos hormonas. Se observaron montas del macho sobre la hembra pero sin constatarse cópulas, las cuales coincidieron con el perfil de HES, que concuerda con lo reportado para el estro en cánidos.

Antimicrobial and Biofilm Inhibitory Activity of Nanoparticles Against Clinical Isolates from Urinary Tract Infection

At these days nanoparticles (NPs) are used as antibacterial due to its have chemical-physical addition to&#x0D; biological effect. A big group of microbial cells adhering to a surface are called biofilm. Exposure to nano&#x0D; particles such as (Ag, Al2O3, Ni) may prevent colonization of new bacteria onto the biofilm. In the present&#x0D; research we have analyze whether the biofilm formation of some isolates of pathogenic bacteria could be&#x0D; influenced by NPs. Also we examined the susceptibility of the isolates to some antibiotics in combination&#x0D; with Ag, Al2O3 and Ni nanoparticles. (50) Strains of isolated K. pneumonia have been examine from patients&#x0D; with urinary tract infection were collected from Education of Baghdad hospitals in Iraq. Serial dilution of&#x0D; Tube method to determination of MIC and MBEC for Nanoparticles against isolates was done. Biofilm&#x0D; formation was evaluated by micro titer plates. Additionally. disc diffusion method was used to assay the&#x0D; various antibiotics and combinations for bactericidal activity against the isolates. The results showed that&#x0D; about 97% of the strains were capable to form of structure biofilms. More than 82% about of strains showed&#x0D; resistance to commonly used antibiotics. Antibiotics and antimicrobials from mixed nanoparticles were&#x0D; higher than compared to nanoparticles alone. Therefore, a synergistic effect was observed between silver -&#x0D; Nickel nanoparticles and silver - aluminum oxide nanoparticles in to inhibiting the growth of K. pneumonia&#x0D; strains. Furthermore, cytotoxicity activity of Ag NPs against Neuro-2a cells was higher than Al2O3 NPs. The&#x0D; results suggest that nanoparticles can be used as antimicrobial agents for therapeutic to treat urinary tract&#x0D; infections caused by K. pneumoniae.

3D Printed Stackable Titer Plate Inserts Supporting Three Interconnected Tissue Models for Drug Transport Studies.

Current in vitro drug screening methods often rely on single-cell models and are therefore imprecise in predicting drug absorption, distribution, metabolism, excretion, and toxicity. This study presents a method to fabricate 3D printed inserts that are compatible with commercially available titer plates. Hydrogels can be casted into the inserts and cells can be cultured either in or on the hydrogels. Once individual cell cultures are fully differentiated, the three different cell cultures are stacked on top of each other for biological experiments. To show the possibilities of this approach, three tissue models representing the first pass metabolism is used. The three tissue models are based on gelatin hydrogels and Caco-2, HUVEC, and HepG2 cells to simulate the small intestine, vascular endothelium, and liver, respectively. The device is simple to fabricate, user friendly, and an alternative to microfluidic-based organ on a chip systems. The presented first pass metabolism study allows for gaining information on drug absorption, distribution, metabolism, and, in the future, excretion in one compact device complying the micro titer plate format.

Interactively AUDIT Your Growth Curves with a Suite of R Packages.

Bottlenecks often occur during data analysis when studying microbial growth in liquid culture at large scale. A researcher can collect thousands of growth curves, repeated measures of a microbial liquid culture, at once in multiple micro titer plates by purpose-built robotic instruments. However, it can be difficult and time-consuming to inspect and analyze these data. This is especially true for researchers without programming experience. To enable this researcher, we created and describe an interactive application: Automated Usher for Data Inspection and Tidying (AUDIT). It allows the user to analyze growth curve data generated from one or more runs each with one or more micro titer plates alongside their experimental design. AUDIT covers input, pre-processing, summarizing, visual exploration and output. Compared to previously available tools AUDIT handles more data, provides live previews and is built from individually re-usable pieces distributed as R packages.

A new sensitive spectrofluorimetric method for measurement of activity and kinetic study of cholinesterases

A new spectrofluorimetric method more sensitive than the Ellman method was developed for determination of both acetylcholinesterase and butyrylcholinesterase activity and for kinetic analysis of these enzymes and their mutants. Two selected mutants of human butyrylcholinesterase (E197Q and E197G) were included in this work. As for the Ellman's method, substrates are thiocholine esters, but the chromogenic reagent, DTNB (dithio-bisnitro benzoic acid) is replaced by a fluorogenic probe, “Calbiochem Probe IV”, (3-(7-Hydroxy-2-oxo-2H-chromen-3-ylcarbamoyl)acrylic acid methylester).Compared to the classical Ellman's method, the sensitivity of this new spectrofluorimetric assay is 2 orders of magnitude higher. The method allows measurement of activity in media containing <10−11 M of cholinesterase active sites at low substrate concentrations, either under first order conditions, [S] << Km, or under conditions where kinetics obeys the Michaelis-Menten model, i.e. at [S] < 1 mM for wild-type enzymes. The method adapted to titration plate reader assays is suitable for clinical and toxicological routine analyses, for high throughput screening of novel cholinesterase mutants and screening of inhibitor libraries of pharmacological interest.

Agile Development of a Microtiter Plate in an Interdisciplinary Project Team

AbstractThe industry is currently changing rapidly. Both customers and employees are focusing much more on their own needs. On the one hand, this requires individualized products and, on the other hand, development processes need to be aligned not only more efficiently but also more closely to the needs of employees. Agile development combines these two characteristics and the second point can be further improved through analyses for collaboration. This is not only necessary for consumer products, but also in medical technology, more and more individualized solutions are required to better help patients. This is also the case with the examination of cells using micro titer plates, which is the subject of this project. Due to the interaction of research activities both on the process and on the product side, this paper presents research results regarding agile product development and collaboration analysis of physical products on the one hand and research results regarding additive and biocompatible production of microtitration plates on the other hand.

Stable and Photothermally Efficient Antibody-Covered Cu3(PO4)2@Polydopamine Nanocomposites for Sensitive and Cost-Effective Immunoassays.

Polydopamine (PDA)-coated or encapsulating Cu3(PO4)2 (Cu3(PO4)2@PDA) nanosheets were synthesized, allowing the C-reaction protein (CRP) antibody to be attached electrostatically for immunosensing of CRP with simple photothermal detection. The antibody-covered Cu3(PO4)2@PDA nanosheets replace the antibody-conjugated enzyme in the enzyme-linked immunosorbant assays. Owing to the high surface area of the 2-D-structured Cu3(PO4)2@PDA nanosheets and the coabsorption of light in the near-IR spectrum by Cu3(PO4)2 and PDA, a small amount of Cu3(PO4)2@PDA confined in the wells of a titer plate generates an easily detectable temperature change after irradiation at 808 nm. The temperature changes, measured by an inexpensive pen-type thermometer, increased linearly with the analyte concentration from 0.42 to 16 pM. We found that the linear relationship can be fitted by the isotherm derived from responses collected from heterogeneous sensors covered with different ligand or antibody densities. The low detection limit (0.11 pM) is largely due to the attachment of a great number of antibodies onto the flat nanosheets. The antibody-covered Cu3(PO4)2@PDA nanosheets are stable and can be used under conditions that are generally unfavorable to enzymatic activities. The excellent agreement between our results and immunoturbidimetric assays of CRP in serum samples from patients and healthy donors demonstrates its utility for disease diagnosis in clinical settings. This cost-effective, biocompatible, and convenient photothermal immunosensor affords a range of possibilities for detecting diverse protein biomarkers.

Determinación de hormonas esteroides en heces de borrego cimarrón (Ovis canadensis Shaw) silvestres en Baja California, México

Objetivo: Determinación de hormonas esteroides en heces de borrego cimarrón silvestre mediante un método no invasivo.&#x0D; Diseño / metodología / enfoque: En este estudio se detectaron progestágenos, andrógenos y estradiol en heces de borrego cimarrón silvestres que habitan en la Sierra San Felipe en Baja California, durante la primavera, invierno y verano del año 2007. Los esteroides se cuantificaron mediante inmunoensayo enzimático y la cuantificación se realizó en una placa de micro titulación.&#x0D; Resultados: Con el perfil hormonal se identificó que 96 eran heces provenientes de machos y 65 de hembras. La concentración de progestágenos en las heces identificadas como hembras varió entre las estaciones del año siendo la primavera con la concentración más alta (280 ng/gr). Las concentraciones de progestágenos y estrógenos en las hembras aumentaron a medida que las condiciones ambientales mejoraron desde el invierno hasta la primavera, coincidiendo con el aumento de andrógenos en los machos de 23.78 ng / gr.&#x0D; Limitaciones en el estudio / implicaciones: con este método no fue posible asignar el sexo de las muestras que se identificaron que provienen de individuos juveniles. Se recomienda analizar muestras de heces frescas para relacionar las concentraciones de hormonas con eventos reproductivos estacionales.&#x0D; Hallazgos / conclusiones: El aumento de las concentraciones de progestágenos y estrógenos en las heces de las hembras y los andrógenos en los machos en la primavera sugiere el inicio de la actividad reproductiva. Las concentraciones de estas hormonas en ambos sexos durante el verano son muy bajas (&lt;5 ng / gr), lo que sugiere una actividad reproductiva baja.&#x0D;

Application of Silver Oxide Nanoparticles as Antibiofilm and Detection of Beta Lactamase Gene in Escherichia Coli Isolated from Diabetic Mellitus Patients

Wound and urine samples of a total of 124 patients with diabetes admitted to the Erbil Teaching and Layla health center in 2017 were inspected, and 80 patients were identified with E.coli. The identification was performed through evaluation of colony appearance, morphological properties, biochemical methods, and the VITEK 2 system. In addition to these, biofilm formation properties were also evaluated using three different methods, namely the micro titer plate, Congo red, and air liquid. For antibiofilm, all micro titer-plate isolated samples were treated with 100μg/ml AgO nanoparticles and the measurements were made with ELISA reader at 630nm. VITEK 2 system was used to test the antibiotic susceptibility of the E.coli samples, using a total of 16 antibiotics. blaSHV and blaCMY genes series were amplified used in determination through PCR test. Identification reference/electrophoresis sequencing identifications revealed 100% match for all isolates of E.coli. Finally, silver nanoparticles were used as the antibacterial agent for the antibiofilm formation tests.

Evaluation of anticancer activity of Melaleuka alternifolia (i. e. tea tree oil) on laryngeal cancer cell line (Hep2) - an in-vitro study

Introduction and Aims A naturally available plant extract of Melaleuka alternifolia tea tree oilTTO is used as a very good antibacterial antifungal antiviral antiprotozoal and anti-inflammatory agent. Currently lot of importance is given for its anticancer effect. Our aim was to evaluate anticancer activity of Melaleuka alternifolia against laryngeal cancer cell line Hep2.Methodology Two different cell lines were used to know the cell viability and cell lysis cytotoxicity in laryngeal cancer Hep2 and monkey kidney cell line Vero used as control. Cell culture The cell lines were maintained in 96 wells micro titer plate containing MEM media supplemented with 10 heat inactivated fetal calf serum FCS containing 5 of mixture of Gentamicin 10ug Penicillin 100 Unitsml and Streptomycin 100microgml in presence of 5 CO2 at 37ordmC for 48-72 hours. Cytotoxicity Assay In vitro growth inhibition effect of test compound was assessed by calorimetric or spectrophotometric determination of conversion of MTT intonbsp Formazan blue by living cells.Results The results represent the mean of five readings. The concentration at which the OD of treated cells was reduced by 50 i.e IC50 value is 0.781microgml of TTO with respect to the untreated control. The standard formula was used for the calculation of cell viability and cell lysis. showed Spearmans Correlation Coefficient Correlation with P value lt0.001 indicating significant results when TTO was treated with Hep2 and Vero cell line. Conclusion TTO is having very good anticancer property against laryngeal cancer cell line Hep2. There was no cytotoxicity noted with Monkey Kidney Vero cell line. The current study can be brought to the level of clinical trials which can show TTO with its greater efficacy related to anticancer activity.

Biofilm Formation by Acinetobacter Spp. in association with Antibiotic Resistance in clinical samples Obtained from Tertiary Care Hospital

Acinetobacter spp., is an opportunistic, gram negative and non-fermenter organism, has occurred as one of the maximum bothersome pathogens for health care organisations worldwide. This organism generally targets the most susceptible hospitalized patients causing pneumonia mainly in patients on mechanical ventilation in ICU, having urinary tract infection, bloodstream infections, skin infections etc. It accounts for about 10% of all nosocomial infections. Its clinical significance over last 15 years has been forced by its amazing capacity to get resistant elements, making it one of the antagonistic organisms in the current antibiotic era. Most of Acinetobacter strains are resistant to all known antibiotics have been reported and they are acting in interaction with this emerging resistance profile, with this ability. Acinetobacter isolatesare able to survive for long periods throughout the hospital environment.Itrecurrently causes infections associated with medical devices, e.g. vascular catheters, cerebrospinal fluid shunts, urinary catheter or an endotracheal tube. Microbial biofilms are considered as virulence factors and is a well-known pathogenic mechanism in such infections. The ability to form biofilms on biotic or abiotic surfaces is a common trait among Acinetobacter strains. Therefore, the present study was undertaken to study antibiotic resistance patterns and association the resistance profile with bio-film production in various clinical isolates of Acinetobacter spp. A total of 35 isolates of Acinetobacter species were obtained for study from various clinical specimens like pus, urine, sputum, blood culture specimens from the patients admitted in hospital. Isolates obtained from different medical specimens were managed and confirmed by conservative microbiological procedures. The isolates of Acinetobacter are identified on the basis of certain biochemical test like catalase positive, oxidase negative, urease negative, nitrate negative, indole negative and non-motility test. A total of 35 isolates were subjected to susceptibility testing by disc diffusion method according to CLSI guidelines for 14 clinically relevant antibiotics. Screening for biofilm production was done quantitatively by micro titre plate assay method. In the present study, 75 % isolates were showing antibiotic resistance and 22.8% isolates produced bio film along with antibiotic resistance. Biofilm formers isolates showed high resistance to imipenem, cephotaxime, ceftazidime, tobramycin, ciprofloxacin as compared to non-biofilm formers. We selected to study the biofilm formation and antibiotic resistance of clinical isolates of Acinetobacter species to understand its ability to persist in the hospital environment to cause outbreaks.