Agar Surface Research Articles

Detection of Bacterial Contamination in Nanoparticle Formulations by Agar Plate Test.

Bacterial contamination can confound the results of in vitro and in vivo preclinical tests. This protocol describes a procedure for detection of microbial contamination in nanotechnology-based formulations. Nanoparticle samples and controls are spread on the surface of agar and growth of bacterial colonies is monitored after 72h of incubation. The intended purpose of this assay is to avoid introduction of microbial contamination into in vitro cell cultures and in vivo animal studies utilizing the test nanomaterial. This assay is not intended to certify the material as sterile.

Genome-Wide Quantitative Fitness Analysis (QFA) of Yeast Cultures.

We provide a detailed protocol for robot-assisted, genome-wide measurement of fitness in the model yeast Saccharomyces cerevisiae using Quantitative Fitness Analysis (QFA). We first describe how we construct thousands of double or triple mutant yeast strains in parallel using Synthetic Genetic Array (SGA) procedures. Strains are inoculated onto solid agar surfaces by liquid spotting followed by repeated photography of agar plates. Growth curves are constructed and the fitness of each strain is estimated. Robot-assisted QFA, can be used to identify genetic interactions and chemical sensitivity/resistance in genome-wide experiments, but QFA can also be used in smaller scale, manual workflows.

Potential anti-leptospiral compound, leptomycin B from marine Streptomyces indiaensis MSU5: taxonomy, fermentation, compound isolation, in vitro and in vivo efficacy

Leptospirosis is a worldwide reemerging tropical zoonotic disease with symptoms of mild febrile illness to more severe multiple organ failure caused by pathogenic leptospiral strains. There was no effective antibiotic for treating leptospirosis. Here, the anti-leptospiral potential of marine actinobacterial compound from Streptomyces indiaensis MSU5 isolated from Manakudy marine sediment, Tamil Nadu, India was evaluated. The potential actinobacterial strain was identified by phenotypic, cell wall, 16S rRNA gene sequence and phylogenetic analysis. In vitro anti-leptospiral activity of the actinobacterial compound was determined using broth microdilution test against various serovars of Leptospira with different concentration ranging from 15.625 to 500µg/ml. Mass production of anti-leptospiral compound was carried out in agar surface fermentation with optimized condition and purified by preparative TLC. The purified fraction of anti-leptospiral compound named as MSU5-1, and it was confirmed by microdilution test. Remarkably, the compound MSU5-1 showed minimum inhibitory concentration of 62.5µg/ml and minimum bactericidal concentration of 125µg/ml against human pathogenic leptospiral isolate strain N2. The structural elucidation of purified compound was carried out using UV, FT-IR, NMR and LC-MS analysis. The compound MSU5-1 was tentatively identified as leptomycin B (C33H48O6) with molecular weight 541.1g/mol. Anti-leptospiral activity of compound MSU5-1 exhibited 80% of survival rate in mice model, further it was confirmed by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) analysis. From the available literature, this is the first report on the marine actinobacterial compound for evaluating both in vitro and in vivo leptospiricidal activity.

Draft Genome Sequence of the Iridescent Marine Bacterium Cellulophaga lytica CECT 8139.

ABSTRACTSome species of the genus Cellulophaga have been reported as having biotechnological interests and noteworthy physiological properties. We report here the draft genome sequence of Cellulophaga lytica CECT 8139, a bacterium that produces an intensely iridescent colony biofilm on agar surfaces.

Effect of Sodium Chloride on Surface-Associated Motility of Acinetobacter baumannii and the Role of AdeRS Two-Component System.

Acinetobacter baumannii is an important bacterial pathogen whose resistance to antibiotics is a growing concern worldwide. Among a wide array of resistance mechanisms displayed by A. baumannii, energy-dependent efflux of antibiotics by proteins belonging the resistance-nodulation-division family serves as an important one. AdeABC pump has been shown to be active in various clinical isolates. Regulation of this pump is controlled by the AdeRS two-component system. In this study, we show that the AdeRS system, in addition to modulating A. baumannii's antibiotic susceptibility, also plays a role in biofilm formation as well as surface-associated motility. We also show that AdeRS deletion mutant is more sensitive to saline stress. In particular, motility of A. baumannii ATCC17978 on agar surface is severely hampered at higher salt concentrations when AdeRS system is absent. Therefore, our study shows that AdeRS could be part of the A. baumannii adaptation strategy to salinity stress.

Comparison of five methods for isolation of DNA from Mycoplasma cynos

Extraction of DNA from Mycoplasma cultured on agar medium is difficult because the plasticity of these microorganisms enables agar penetration. This eventually causes cell loss during harvesting of colonies from the agar surface. Here, we used the GenElute™ gel extraction kit, which is usually used to purify polymerase chain reaction products, for extracting DNA from Mycoplasma. We compared the DNA extraction efficiency of the GenElute™ gel extraction kit from Mycoplasma cynos cultured in agar medium with four other DNA extraction methods. The results were evaluated based on the purity and amount of DNA obtained from one Mycoplasma colony. Eight strains of Mycoplasma cynos isolated from the broncho-alveolar lavage fluid of dogs were used. The GenElute™ gel extraction protocol was the most efficient among all the methods tested in this study as it yielded the highest amount and the purest quality of DNA (199.3±0.744ng/μl) from a single colony. Among the methods tested, the GenElute™ gel extraction method is the most rapid, sensitive, and simple method for DNA extraction from Mycoplasma. This procedure may also prove useful for extracting DNA from other Mycoplasma species.

Inactivation of foodborne pathogens on food packaging and in cow milk by exposure to a Nd:YAG laser

This study demonstrates the inhibition of selective foodborne pathogenic bacteria by exposure to a 355 and 266 nm Nd:YAG laser. In baseline assay P. aeruginosa showed maximum inhibition followed by Escherichia coli, Staphylococcus aureus, Salmonella typhimurium, and Listeria monocytogenes grown on agar surface. Cell lysis was confirmed by confocal microscopy for all pathogens. In liquid media, P. aeruginosa was irradiated with the third harmonic of a Nd:YAG laser pulse energy of 185 mJ, which was able to reduce the population of 1.65 × 105 by 4.7 logs, while P. aeruginosa could only be reduced by 0.73 log/cm2 on packaging material surface. In the final phase, the laser’s potential was preliminarily tested for sterilization of raw cow milk. The reduction patterns of E. coli, Salmonella sp. yeasts and Lactobacillus sp. were 30%, 25%, 47%, and 30%, respectively, in raw milk, moreover, laser exposure had no significant impact on physio-chemistry of milk. Therefore, results indicate the potential application of laser in packing materials and milk sterilization at the industrial level.

Comparative genome and transcriptome analysis reveals distinctive surface characteristics and unique physiological potentials of Pseudomonas aeruginosa ATCC 27853

BackgroundPseudomonas aeruginosa ATCC 27853 was isolated from a hospital blood specimen in 1971 and has been widely used as a model strain to survey antibiotics susceptibilities, biofilm development, and metabolic activities of Pseudomonas spp.. Although four draft genomes of P. aeruginosa ATCC 27853 have been sequenced, the complete genome of this strain is still lacking, hindering a comprehensive understanding of its physiology and functional genome.ResultsHere we sequenced and assembled the complete genome of P. aeruginosa ATCC 27853 using the Pacific Biosciences SMRT (PacBio) technology and Illumina sequencing platform. We found that accessory genes of ATCC 27853 including prophages and genomic islands (GIs) mainly contribute to the difference between P. aeruginosa ATCC 27853 and other P. aeruginosa strains. Seven prophages were identified within the genome of P. aeruginosa ATCC 27853. Of the predicted 25 GIs, three contain genes that encode monoxoygenases, dioxygenases and hydrolases that could be involved in the metabolism of aromatic compounds. Surveying virulence-related genes revealed that a series of genes that encode the B-band O-antigen of LPS are lacking in ATCC 27853. Distinctive SNPs in genes of cellular adhesion proteins such as type IV pili and flagella biosynthesis were also observed in this strain. Colony morphology analysis confirmed an enhanced biofilm formation capability of ATCC 27853 on solid agar surface compared to Pseudomonas aeruginosa PAO1. We then performed transcriptome analysis of ATCC 27853 and PAO1 using RNA-seq and compared the expression of orthologous genes to understand the functional genome and the genomic details underlying the distinctive colony morphogenesis. These analyses revealed an increased expression of genes involved in cellular adhesion and biofilm maturation such as type IV pili, exopolysaccharide and electron transport chain components in ATCC 27853 compared with PAO1. In addition, distinctive expression profiles of the virulence genes lecA, lasB, quorum sensing regulators LasI/R, and the type I, III and VI secretion systems were observed in the two strains.ConclusionsThe complete genome sequence of P. aeruginosa ATCC 27853 reveals the comprehensive genetic background of the strain, and provides genetic basis for several interesting findings about the functions of surface associated proteins, prophages, and genomic islands. Comparative transcriptome analysis of P. aeruginosa ATCC 27853 and PAO1 revealed several classes of differentially expressed genes in the two strains, underlying the genetic and molecular details of several known and yet to be explored morphological and physiological potentials of P. aeruginosa ATCC 27853.

Transcriptomic Analysis of the Swarm Motility Phenotype of Salmonella enterica Serovar Typhimurium Mutant Defective in Periplasmic Glucan Synthesis.

Movement of food-borne pathogens on moist surfaces enables them to migrate towards more favorable niches and facilitate their survival for extended periods of time. Salmonella enterica serovar Typhimurium mutants defective in Osmoregulated periplasmic glucans (OPG) synthesis are unable to exhibit motility on moist surfaces (swarming); however, their mobility in liquid (swim motility) remains unaffected. In order to understand the role of OPG in swarm motility, transcriptomic analysis was performed using cells growing on a moist agar surface. In opgGH deletion mutant, lack of OPG significantly altered transcription of 1039 genes out of total 4712 genes (22%). Introduction of a plasmid-borne copy of opgGH into opgGH deletion mutant restored normal expression of all but 30 genes, indicating a wide-range influence of OPG on gene expression under swarm motility condition. Major pathways that were differentially expressed in opgGH mutants were motility, virulence and invasion, and genes related to the secondary messenger molecule, cyclic di-GMP. These observations provide insights and help explain the pleiotropic nature of OPG mutants such as sub-optimal virulence and competitive organ colonization in mice, biofilm formation, and sensitivity towards detergent stress.

Bioavailability of Soil-Sorbed Tetracycline to Escherichia coli under Unsaturated Conditions.

Increasing concentrations of anthropogenic antibiotics in soils are partly responsible for the proliferation of bacterial antibiotic resistance. However, little is known about how soil-sorbed antibiotics exert selective pressure on bacteria in unsaturated soils. This study investigated the bioavailability of tetracycline sorbed on three soils (Webster clay loam, Capac sandy clay loam, and Oshtemo loamy sand) to a fluorescent Escherichia coli bioreporter under unsaturated conditions using agar diffusion assay, microscopic visualization, and model simulation. Tetracycline sorbed on the soils could be desorbed and become bioavailable to the E. coli cells at matric water potentials of -2.95 to -13.75 kPa. Bright fluorescent rings were formed around the tetracycline-loaded soils on the unsaturated agar surfaces, likely due to radial diffusion of tetracycline desorbed from the soils, tetracycline uptake by the E. coli cells, and its inhibition on E. coli growth, which was supported by the model simulation. The bioavailability of soil-sorbed tetracycline was much higher for the Oshtemo soil, probably due to faster diffusion of tetracycline in coarse-textured soils. Decreased bioavailability of soil-sorbed tetracycline at lower soil water potential likely resulted from reduced tetracycline diffusion in soil pore water at smaller matric potential and/or suppressed tetracycline uptake by E. coli at lower osmotic potential. Therefore, soil-sorbed tetracycline could still exert selective pressure on the exposed bacteria, which was influenced by soil physical processes controlled by soil texture and soil water potential.

Increased Biological Activity of Aneurinibacillus migulanus Strains Correlates with the Production of New Gramicidin Secondary Metabolites

The soil-borne gram-positive bacteria Aneurinibacillus migulanus strain Nagano shows considerable potential as a biocontrol agent against plant diseases. In contrast, A. migulanus NCTC 7096 proved less effective for inhibition of plant pathogens. Nagano strain exerts biocontrol activity against some gram-positive and gram-negative bacteria, fungi and oomycetes through the production of gramicidin S (GS). Apart from the antibiotic effects, GS increases the rate of evaporation from the plant surface, reducing periods of surface wetness and thereby indirectly inhibiting spore germination. To elucidate the molecular basis of differential biocontrol abilities of Nagano and NCTC 7096, we compared GS production and biosurfactant secretion in addition to genome mining of the genomes. Our results proved that: (i) Using oil spreading, blood agar lysis, surface tension and tomato leaves wetness assays, Nagano showed increased biosurfactant secretion in comparison with NCTC 7096, (ii) Genome mining indicated the presence of GS genes in both Nagano and NCTC 7096 with two amino acid units difference between the strains: T342I and P419S. Using 3D models and the DUET server, T342I and P419S were predicted to decrease the stability of the NCTC 7096 GS synthase, (iii) Nagano produced two additional GS-like molecules GS-1155 (molecular weight 1155) and GS-1169 (molecular weight 1169), where one or two ornithine residues replace lysine in the peptide. There was also a negative correlation between surface tension and the quantity of GS-1169 present in Nagano, and (iv) the Nagano genome had a full protein network of exopolysaccharide biosynthesis in contrast to NCTC 7096 which lacked the first enzyme of the network. NCTC 7096 is unable to form biofilms as observed for Nagano. Different molecular layers, mainly gramicidin secondary metabolite production, account for differential biocontrol abilities of Nagano and NCTC 7096. This work highlighted the basis of differential biological control abilities between strains belonging to the same species and demonstrates techniques useful to the screening of effective biocontrol strains for environmentally friendly secondary metabolites that can be used to manage plant pathogens in the field.

Influence of Physical Effects on the Swarming Motility of Pseudomonas aeruginosa

Many species of bacteria can spread over a moist surface via a particular form of collective motion known as “surface swarming”. This form of motility is typically studied by inoculating bacteria on a gel formed by 0.4–1.5% agar, which contains essential nutrients for their growth and proliferation. Using Pseudomonas aeruginosa and its pili-less mutant, ΔPilA, we investigate physical factors that either facilitate or restrict the swarming motility, measured by the rate of increase in area covered by a spreading bacterial colony, i.e., a swarm. The wild-type colony spreads over the agar surface in highly branched structures. The pili-less mutant fills up the area more fully as it spreads, but it also produces numerous and fragmented branches, or tendrils, at the swarm front. Whereas additional surfactants enhance swarming, increasing the agar percentage, adding extra salt or sugar or incorporating viscous agents in the agar matrix all decrease swarming, supporting the conclusion that swarming motility is restricted by the surface tension at the swarm front and swarm growth is limited by the rate of water supply from within the agar gel. The physical basis elaborated through this study provides a useful framework for understanding the swarming behavior of numerous species of bacteria.

FIRST REPORT OF LEAF SPOT CAUSED BY COLLETOTRICHUM COCCODES ON VIOLA TRICOLOR IN IRAN

Viola L. is the largest genus of Violaceae. Violets naturally grow in northern Iran, but they are cultivated in most parts of Iran. The fungus Colletotrichum coccodes is a common pathogen on vegetables of Solanaceae family, causing fruit anthracnose of tomato, pepper and eggplant and black dot of potato (Rodeva et al., 2016). Viola sp. from West Azerbaijan is a new host for Colletotrichum sp. in the C. destructivum species complex (Arzanlou et al., 2015). During a survey in 2016 on the campus of Sari Agricultural University, symptoms of leaf spot, necrotic rings with a tan center and dark brown margin were observed on violets. Isolation and purification of suspected fungal agent were done on WA medium, then transferred to Potato dextrose agar (PDA). . Colonies of Colletotrichum sp. grown on PDA plates were dominated by white, sparse aerial mycelium, which becomes grey with age, and by abundant, black sclerotia evenly distributed over the agar surface. Conidia were 16–24×3–4 µm, straight, fusiform and abruptly tapered to each end. For pathogenicity tests leaves were inoculated with a mycelial plug from the periphery of the representative isolate (C4a) grown on PDA. After 10-15 days, symptoms similar to those of the original leaves were observed. The fungus was reisolated on PDA from the lesions on the inoculated leaves; the colonies and the morphology of conidia were the same as those of the original isolates. For molecular identification, the internal transcribed spacer (ITS) regions of isolate C4a were amplified with the universal ITS4/ITS5 primers (White et al., 1990). The 600 bp product was sequenced (GenBank accession No. KY176353) and showed the highest similarity (99%) with C. coccodes (AJ301984.1) To our knowledge, this is the first report of C. coccodes in Iran causing leaf spot on Viola tricolor.

Predicting path from undulations for C. elegans using linear and nonlinear resistive force theory

A basic issue in the physics of behaviour is the mechanical relationship between an animal and its surroundings. The model nematode C. elegans provides an excellent platform to explore this relationship due to its anatomical simplicity. Nonetheless, the physics of nematode crawling, in which the worm undulates its body to move on a wet surface, is not completely understood and the mathematical models often used to describe this phenomenon are empirical. We confirm that linear resistive force theory, one such empirical model, is effective at predicting a worm’s path from its sequence of body postures for forward crawling, reversing, and turning and for a broad range of different behavioural phenotypes observed in mutant worms. Worms recently isolated from the wild have a higher effective drag anisotropy than the laboratory-adapted strain N2 and most mutant strains. This means the wild isolates crawl with less surface slip, perhaps reflecting more efficient gaits. The drag anisotropies required to fit the observed locomotion data (70 ± 28 for the wild isolates) are significantly larger than the values measured by directly dragging worms along agar surfaces (3–10 in Rabets et al (2014 Biophys. J. 107 1980–7)). A proposed nonlinear extension of the resistive force theory model also provides accurate predictions, but does not resolve the discrepancy between the parameters required to achieve good path prediction and the experimentally measured parameters. We confirm that linear resistive force theory provides a good effective model of worm crawling that can be used in applications such as whole-animal simulations and advanced tracking algorithms, but that the nature of the physical interaction between worms and their most commonly studied laboratory substrate remains unresolved.

Candida krusei form mycelia along agar surfaces towards each other and other Candida species

BackgroundCandida krusei has been known to exhibit communal interactions such as pellicle formation and crawling out of nutritional broth. We noticed another possible interaction on agar surfaces, where C. krusei yeast cells formed mycelia along agar surfaces toward each other. We report here the results of experiments to study this interaction.ResultsWhen C.krusei yeast cells are plated in parallel streaks, they form mycelia along agar surfaces toward other yeasts. They also detect the presence of Candida albicans and Candida glabrata across agar surfaces, while the latter two react neither to their own kind, nor to C. krusei. Secreted molecule(s) are likely involved as C.krusei does not react to heat killed C. krusei. Timing and rate of mycelia formation across distances suggests that mycelia start forming when a secreted molecule(s) on agar surface reaches a certain concentration. We detected farnesol, tyrosol and tryptophol molecules that may be involved with mycelial formation, on the agar surfaces between yeast streaks. Unexpectedly the amounts detected between streaks were significantly higher than would have expected from additive amounts of two streaks. All three Candida species secreted these molecules. When tested on agar surface however, none of these molecules individually or combined induced mycelia formation by C. krusei.ConclusionsOur data confirms another communal interaction by C. krusei, manifested by formation of mycelia by yeast cells toward their own kind and other yeasts on agar surfaces. We detected secretion of farnesol, tyrosol and tryptophol by C. krusei but none of these molecules induced this activity on agar surface making it unlikely that they are the ones utilized by this yeast for this activity.

Impact of Salt and Nutrient Content on Biofilm Formation by Vibrio fischeri.

Vibrio fischeri, a marine bacterium and symbiont of the Hawaiian bobtail squid Euprymna scolopes, depends on biofilm formation for successful colonization of the squid’s symbiotic light organ. Here, we investigated if culture conditions, such as nutrient and salt availability, affect biofilm formation by V. fischeri by testing the formation of wrinkled colonies on solid media. We found that V. fischeri forms colonies with more substantial wrinkling when grown on the nutrient-dense LBS medium containing NaCl relative to those formed on the more nutrient-poor, seawater-salt containing SWT medium. The presence of both tryptone and yeast extract was necessary for the production of “normal” wrinkled colonies; when grown on tryptone alone, the colonies displayed a divoting phenotype and were attached to the agar surface. We also found that the type and concentration of specific seawater salts influenced the timing of biofilm formation. Of the conditions assayed, wrinkled colony formation occurred earliest in LBS(-Tris) media containing 425 mM NaCl, 35 mM MgSO4, and 5 mM CaCl2. Pellicle formation, another measure of biofilm development, was also enhanced in these growth conditions. Therefore, both nutrient and salt availability contribute to V. fischeri biofilm formation. While growth was unaffected, these optimized conditions resulted in increased syp locus expression as measured by a PsypA-lacZ transcriptional reporter. We anticipate these studies will help us understand how the natural environment of V. fischeri affects its ability to form biofilms and, ultimately, colonize E. scolopes.

Quantitative Determination of Anti-bacterial Activity During Bacterial Co-culture.

Anti-bacterial activity assays are an important tool in the assessment of the ability of one bacterium to kill or inhibit the growth of another, for example during the study of the Type VI secretion system (T6SS) and the anti-bacterial toxins it secretes. The method we describe here can detect the ability of a bacterial strain to kill or inhibit other bacterial cells in a contact-dependent manner when co-cultured on an agar surface. It is particularly useful since it enumerates the recovery of viable target cells and thus enables quantification of the anti-bacterial activity. We provide a detailed description of how to measure the T6SS-dependent anti-bacterial activity of a bacterium such as Serratia marcescens against a competitor prokaryotic organism, Escherichia coli, and also describe possible variations in the method to allow adaptation to other attacker and target organisms.

Good Bugs vs Bad Bugs: Evaluation of Inhibitory Effect of Selected Probiotics against Enterococcus faecalis.

The main goal of endodontics is the prevention of apical periodontitis. This is due to the presence of persistent pathogenic microorganisms, such as Enterococcus faecalis, and its ability to directly cause acute and chronic inflammation in the periapical tissues. Lactobacillus has been shown to promote health in the intestines as well as to inhibit the growth of certain problematic oral bacteria. This study explores shifting the established paradigm of endodontic treatment, which has focused on eliminating all bacteria from the canal system and on elimination of the problematic bacteria through introducing probiotics. A preliminary work was performed to evaluate the possible effectiveness of probiotics in preventing the growth of E. faecalis. Two probiotic cultures Lactobacillus plantarum ATCC 8041 and Lactobacillus rhamnosus ATCC 7408 were selected to check their antimicrobial activity against E. faecalis ATCC 29212 by two methods: agar cup/well diffusion method and deferred antagonism test. Agar cup method: A total of 0.5 mL of requisite test pathogen culture was inoculated into 20 mL of molten sterile Mueller and Hinton agar and cooled to 45 ± 2°C. Circular wells of diameter 10 mm were punched in each of the poured plates. Appropriately diluted test samples were added to the above-punched wells. The plates were incubated upright position at 37°C for 24 hours in aerobic conditions. Postincubation, zone of inhibition was measured. The cell-free supernatant of Lactobacillus species was also evaluated for antimicrobial activity. Deferred antagonism test: The test probiotic strain was standardized to 0.1 optical density (OD) at 600 nm and inoculated in a 1 cm wide diametric streak across the surface of trypticase soy agar + yeast extract + calcium carbonate (TSYCa) agar using a sterile cotton swab. Then, the plate agar containing the test strain was incubated at 37°C for 24 hours under aero-philic condition and then standardized to 0.1 OD at 600 nm for overnight (18 hours, 37°C). A purified culture of indicator strain (pathogen) was streaked at right angles to the line of original producer growth. Postincubation plates were observed for the inhibition zone width of the indicator strain. Under the conditions of this study, Lactobacilli had an inhibitory effect on the growth of E. faecalis by agar cup method but not by deferred antagonism test. This pilot study demonstrated that probiotics show a potential in root canal therapy. If probioitcs are effective against end-odontic pathogens, they can be potentially used as intracanal medicaments. This will be a novel concept of introducing bac-teriotherapy in endodontics and replacing pathogenic bacteria by healthy bacteria, normal flora.

The effect of sodium bicarbonate oral rinse on salivary pH and oral microflora: A prospective cohort study.

Aim:Present study is designed to explore the effect of sodium bicarbonate oral rinse on salivary pH and oral micro flora.Materials and Methods:Twenty five healthy subjects were recruited for the study in department of dentistry in Era Medical College. Subjects were abstained from tooth brushing overnight pre rinse (control) samples were collected after one hour of dinner and were asked to rinse with pre calibrated freshly prepared sodium bicarbonate solution. The salivary samples were then collected the following morning using sterile gauze in marked bottles. Aerobic bacterial culture was done by plating the sample directly from the swab on the surface of Blood agar and Mac Conkeys media respectively. The colony forming units and ph were calculated for the pre rinse and post rinse saliva sample.Result:Results shows that salivary pH increased significantly after sodium Bicarbonate oral rinse. There was a marginal decrease in number of CFU/ml for bacteria especially Viridans Streptococci, Moraxella species.Conclusion:Sodium Bicarbonate oral rinse may be considered as a cheap and effective alternative for chlorhexidine and alcohol based mouth wash, especially where long duration usage is required.

Comparison of Normal Resident Flora on the Face of Medical Students who use and who do not use Cosmetics

Introduction: Normal flora of the skin plays a beneficial role in preventing the pathogenic organisms from colonizing the skin and causing infection. It is possible that the facial cosmetics may cause a change in the normal flora disrupting its protective function. Aim: To find out the effect of cosmetics, those that are applied on to the face on resident normal flora of the face. Materials and Methods: This was a cross-sectional study involving medical students aged 19 to 25 years, of whom 42 were regular cosmetic users and another 42 were non cosmetic users. Swabs were taken from the face of the subjects after a face wash with soap and water and eluted in 1 ml sterile peptone water. Tenfold dilutions of the sample were made and 100 μl of the diluted sample was spread over the surface of Trypticase soy agar, Mac Conkey’s agar and blood agar. The colonies were counted and also identified. Statistical evaluation was done by Chi square test using SPSS version 16. R esults: We isolated Coagulase Negative Staphylococci (CoNS), Micrococcus spp. and methicillin sensitive S.aureus, non-fermenting oxidase negative gram negative bacilli and diphtheroids from both the groups. We found that most of the students who were regular cosmetic users 22 (52.38%) yielded negligent growth in comparison with non-cosmetic users (16, 38%). CoNS was less frequently isolated from regular cosmetic users 10 (23.8%) in comparison with those who do not use cosmetics 17 (40.47%). Conclusion: It is possible that regular use of cosmetics does reduce the normal flora but further studies with larger sample size are required to prove and confirm this finding.