Beasley J, Craig B, DeBrota M, McClure W - Isolation and Classification of Soil Microorganisms with Suspected Antimicrobial Properties - Project Poster
As part of the BIO220 Microbiology course at Rose-Hulman Institute of Technology, our group conducted a study isolating and classifying various soil microorganisms with suspected antimicrobial properties.
Microbiological Quality Assessment of Cereal Based Food ‘Pitha’ On Retail Sal...IOSR Journals
Pitha‟ is one of the most popular traditional foods made up from various kinds of cereal grains like rice, wheat‟s, flours etc. The microbiological quality of pitha on retail sale in the street of Dhaka city was studied with thirty six different samples by detecting the range of total viable count (TVC) of bacteria, indicator microorganisms, such as Escherichia coli, Enterobacteriaceae and the common food pathogens Staphylococcus aureus, Bacillus cereus., Salmonella Spp. and Pseudomonas spp. In our study, the majority (67.5%) of the microbial contaminants were gram negative rods. The TVC of bacteria was ranged from (9.0x103) cfu/g to (1.2x106) cfu/g. Total coliform, fecal coliform and E. coli were detected from 30.56%, 22.2% and 13.8% of the samples respectively indicating unsatisfactory results. For Salmonella spp. 16.6% of the samples were unable to reach the acceptability levels. Besides, according to the ICMSF (International Commission on Microbiological Specifications for food), in terms of Pseudomonas spp. 33.3%, for Staphylococcus aureus 12.2% and for Bacillus cereus 11.1% of the samples were not suitable for the consumption. All these phenomena indicate poor hygiene practices. This study emphasizes the need to improve production hygiene.
Microbiological Quality Assessment of Cereal Based Food ‘Pitha’ On Retail Sal...IOSR Journals
Pitha‟ is one of the most popular traditional foods made up from various kinds of cereal grains like rice, wheat‟s, flours etc. The microbiological quality of pitha on retail sale in the street of Dhaka city was studied with thirty six different samples by detecting the range of total viable count (TVC) of bacteria, indicator microorganisms, such as Escherichia coli, Enterobacteriaceae and the common food pathogens Staphylococcus aureus, Bacillus cereus., Salmonella Spp. and Pseudomonas spp. In our study, the majority (67.5%) of the microbial contaminants were gram negative rods. The TVC of bacteria was ranged from (9.0x103) cfu/g to (1.2x106) cfu/g. Total coliform, fecal coliform and E. coli were detected from 30.56%, 22.2% and 13.8% of the samples respectively indicating unsatisfactory results. For Salmonella spp. 16.6% of the samples were unable to reach the acceptability levels. Besides, according to the ICMSF (International Commission on Microbiological Specifications for food), in terms of Pseudomonas spp. 33.3%, for Staphylococcus aureus 12.2% and for Bacillus cereus 11.1% of the samples were not suitable for the consumption. All these phenomena indicate poor hygiene practices. This study emphasizes the need to improve production hygiene.
This slide lecture is for students seeking help regarding Metagenomics. Do remember me in your prayers.
Metagenomics Applications, Metagenomics working principles , Metagenomic libraries
, Metagenomic Techniques , Metagenomics limitations and other topics are elaborated in this Slideshare.
Fa1zanS: Methods of studying biochemical properties of microorganismFaizan Siddiqui
Methods of studying biochemical properties of microorganism, how they are cultured in laboratory and how they play key role in maintaining the equilibrium of our environment, their role in energy production, agriculture, ecological balance. Media: Providing Nutrients in the Laboratory. Incubation, Inspection, and Identification. Methods of cultivation
Drug discovery is the process through which potential new medicines are identified.
It involves a wide range of scientific disciplines, including biology, chemistry and pharmacology.
Natural products and their structural analogues have historically made a major contribution to pharmacotherapy, especially for cancer and infectious diseases.
Natural products also have challenges for drug discovery, such as technical barriers to screening, isolation, characterization and optimization.
Metabolomics Analysis on Antifungal Activities Produced by Penicillium oxalic...Agriculture Journal IJOEAR
—In-vitro antagonist tests such as disc diffusion and minimum inhibition concentration (MIC) were conducted against C. gloeosporioides. 1 H-NMR coupled with multivariate statistical analysis was carried out to identify possible compounds produced. Glucose crude extract exhibited the highest percent inhibition of radial growth (PIRG) with 75% and the lowest MIC value with 78 µg mL-1. For metabolomics, different metabolites produced were clustered according to the carbon sources used and gave a representative impression of the metabolites produced by P. oxalicum T3.3. The study has shown the potential of using a combination of 1 H-NMR spectroscopy and multivariate statistical analysis and their correlation with MIC in differentiating the effect of carbon sources used based on the identification of possible metabolites contributing to their differences. Findings from this work may potentially provide the basis for further studies on both antimicrobial activities against plant pathogen and elucidation of the metabolite compounds produced by P. oxalicum T3.3.
Antonelli K, DeBrota M, Durgham T, Kress N, Miller B, Zhang Q - Synthetic Foo...Michael DeBrota
This project was completed as part of Miami University's CHM491Q Chemistry in Societal Issues capstone course toward partial completion of my Bachelor of Science degree in Biochemistry (Oxford, OH, May 2020).
Title:
Synthetic Food Additives: Chemistry, Health Effects, and the Societal Dilemma
Authors:
Kayleigh Antonelli, Michael DeBrota, Tony Durgham, Nicholas Kress, Bryce Miller, Qingxin Zhang
Advisor:
Dr. Michael A. Kennedy, Department of Chemistry and Biochemistry, Miami University
Abstract:
Have you ever been puzzled by complex-sounding ingredients on your food label, wondering why they are used and what they do? Synthetic additives are ubiquitous in today's commercial food products, and their definitions, uses, and regulations are surprisingly complex.
Today, many categories of synthetic additives exist, designed to improve food's characteristics or manufacturing processes. Various principles of chemistry underlie the functioning of these additives, and each possesses a unique and specific function.
Certain additives have been subject to public scrutiny due to suspected negative health effects arising from their consumption. Moreover, the use of additives presents an additional challenge in that the accessibility to healthy alternatives, especially for individuals with low socioeconomic status, is hindered by the widespread availability of cheap, highly processed foods.
Here, we present a summary of information gathered from recent studies, journal publications, and reference sources, with the intent of providing a general overview on synthetic food additives. To this end, we examine their history and definitions, the chemical basis of their function with special focus on three commonly used additives, concerns and benefits surrounding their use, and potential strategies for mitigating these concerns.
DeBrota M, Smith C, Tanis-Arens C - PACU Nursing Education for the Congenital...Michael DeBrota
This is the project abstract submitted to the AMAAC MARC 2023 conference. The project poster can be found here: https://www.slideshare.net/MichaelDeBrota/debrota-m-smith-c-tanisarens-c-pacu-nursing-education-for-the-congenital-cardiac-patient-a-quality-improvement-initiative-project-poster
More Related Content
Similar to Beasley J, Craig B, DeBrota M, McClure W - Isolation and Classification of Soil Microorganisms with Suspected Antimicrobial Properties - Project Poster
This slide lecture is for students seeking help regarding Metagenomics. Do remember me in your prayers.
Metagenomics Applications, Metagenomics working principles , Metagenomic libraries
, Metagenomic Techniques , Metagenomics limitations and other topics are elaborated in this Slideshare.
Fa1zanS: Methods of studying biochemical properties of microorganismFaizan Siddiqui
Methods of studying biochemical properties of microorganism, how they are cultured in laboratory and how they play key role in maintaining the equilibrium of our environment, their role in energy production, agriculture, ecological balance. Media: Providing Nutrients in the Laboratory. Incubation, Inspection, and Identification. Methods of cultivation
Drug discovery is the process through which potential new medicines are identified.
It involves a wide range of scientific disciplines, including biology, chemistry and pharmacology.
Natural products and their structural analogues have historically made a major contribution to pharmacotherapy, especially for cancer and infectious diseases.
Natural products also have challenges for drug discovery, such as technical barriers to screening, isolation, characterization and optimization.
Metabolomics Analysis on Antifungal Activities Produced by Penicillium oxalic...Agriculture Journal IJOEAR
—In-vitro antagonist tests such as disc diffusion and minimum inhibition concentration (MIC) were conducted against C. gloeosporioides. 1 H-NMR coupled with multivariate statistical analysis was carried out to identify possible compounds produced. Glucose crude extract exhibited the highest percent inhibition of radial growth (PIRG) with 75% and the lowest MIC value with 78 µg mL-1. For metabolomics, different metabolites produced were clustered according to the carbon sources used and gave a representative impression of the metabolites produced by P. oxalicum T3.3. The study has shown the potential of using a combination of 1 H-NMR spectroscopy and multivariate statistical analysis and their correlation with MIC in differentiating the effect of carbon sources used based on the identification of possible metabolites contributing to their differences. Findings from this work may potentially provide the basis for further studies on both antimicrobial activities against plant pathogen and elucidation of the metabolite compounds produced by P. oxalicum T3.3.
Similar to Beasley J, Craig B, DeBrota M, McClure W - Isolation and Classification of Soil Microorganisms with Suspected Antimicrobial Properties - Project Poster (20)
Antonelli K, DeBrota M, Durgham T, Kress N, Miller B, Zhang Q - Synthetic Foo...Michael DeBrota
This project was completed as part of Miami University's CHM491Q Chemistry in Societal Issues capstone course toward partial completion of my Bachelor of Science degree in Biochemistry (Oxford, OH, May 2020).
Title:
Synthetic Food Additives: Chemistry, Health Effects, and the Societal Dilemma
Authors:
Kayleigh Antonelli, Michael DeBrota, Tony Durgham, Nicholas Kress, Bryce Miller, Qingxin Zhang
Advisor:
Dr. Michael A. Kennedy, Department of Chemistry and Biochemistry, Miami University
Abstract:
Have you ever been puzzled by complex-sounding ingredients on your food label, wondering why they are used and what they do? Synthetic additives are ubiquitous in today's commercial food products, and their definitions, uses, and regulations are surprisingly complex.
Today, many categories of synthetic additives exist, designed to improve food's characteristics or manufacturing processes. Various principles of chemistry underlie the functioning of these additives, and each possesses a unique and specific function.
Certain additives have been subject to public scrutiny due to suspected negative health effects arising from their consumption. Moreover, the use of additives presents an additional challenge in that the accessibility to healthy alternatives, especially for individuals with low socioeconomic status, is hindered by the widespread availability of cheap, highly processed foods.
Here, we present a summary of information gathered from recent studies, journal publications, and reference sources, with the intent of providing a general overview on synthetic food additives. To this end, we examine their history and definitions, the chemical basis of their function with special focus on three commonly used additives, concerns and benefits surrounding their use, and potential strategies for mitigating these concerns.
DeBrota M, Smith C, Tanis-Arens C - PACU Nursing Education for the Congenital...Michael DeBrota
This is the project abstract submitted to the AMAAC MARC 2023 conference. The project poster can be found here: https://www.slideshare.net/MichaelDeBrota/debrota-m-smith-c-tanisarens-c-pacu-nursing-education-for-the-congenital-cardiac-patient-a-quality-improvement-initiative-project-poster
DeBrota M, Smith C, Tanis-Arens C - PACU Nursing Education for the Congenital...Michael DeBrota
This is the poster presented at the AMAAC MARC 2023 Conference in Milwaukee, WI.
Abstract can be found here: https://www.slideshare.net/MichaelDeBrota/debrota-m-smith-c-tanisarens-c-pacu-nursing-education-for-the-congenital-cardiac-patient-a-quality-improvement-initiative-project-abstract
DeBrota M, Penry O - Isolation and Classification of Soil Microorganisms with...Michael DeBrota
As part of the BIO220 Microbiology course at Rose-Hulman Institute of Technology, our group conducted a study isolating and classifying various soil microorganisms with suspected antimicrobial properties.
DeBrota M, Pflueger J - Studies of Cobalt Boride-Mediated Nitrile Reductions ...Michael DeBrota
Under the auspices of the R-SURF program, I conducted an independent, one-on-one faculty-mentored organic chemistry research project with Dr. Jason J. Pflueger, PhD. My project, partially building on previous research conducted by Sarpong, Pflueger, et al., investigated organic synthesis methods using cobalt boride species. In the course of conducting the various experimental reactions during the project, we unexpectedly discovered a potential novel reaction scheme for the single-step reduction of benzonitrile to benzyl alcohol using greener and more ubiquitous reagents than those used in extant literature. Following the completion of the project, I created a formal laboratory report in addition to this poster, and presented the project’s findings at the Eli Lilly Undergraduate Grantee Symposium in Indianapolis, IN, in August 2018.
DeBrota - The Selective Cobalt Boride-Mediated Reduction of Benzonitrile to B...Michael DeBrota
This report represents the culmination of an independent, one-on-one faculty-mentored organic chemistry research project with Dr. Jason J. Pflueger, PhD, at Rose-Hulman Institute of Technology, under the auspices of the Rose-Hulman Summer Undergraduate Research Fellowship (R-SURF). My project, partially building on previous research conducted by Sarpong, Pflueger, et al., investigated organic synthesis methods using cobalt boride species. In the course of conducting the various experimental reactions during the project, we unexpectedly discovered a potential novel reaction scheme for the single-step reduction of benzonitrile to benzyl alcohol using greener and more ubiquitous reagents than those used in extant literature. Following the completion of the project, I created a formal laboratory report and scientific poster, and presented the project’s findings at the Eli Lilly Undergraduate Grantee Symposium in Indianapolis, IN, in August 2018.
Agarwal A, DeBrota M, Fenimore L - Purification and Quantification of Cytochr...Michael DeBrota
As part of Rose-Hulman’s CHEM291 Introduction to Chemical Research course, this group project entailed the extraction, isolation, and quantification of cytochrome c from gross bovine liver tissue, using an experimental protocol assembled from review of multiple sources of primary literature. Core biochemical laboratory techniques such as tissue extraction, cell lysis, centrifugation, and purification were practiced, along with quantitative methods for confirming experimental yield. Following the completion of the project, our group wrote a formal laboratory report in addition to this scientific poster.
Agarwal A, DeBrota M, Fenimore L - Purification and Quantification of Cytochr...Michael DeBrota
As part of Rose-Hulman’s CHEM291 Introduction to Chemical Research course, this group project entailed the extraction, isolation, and quantification of cytochrome c from gross bovine liver tissue, using an experimental protocol assembled from review of multiple sources of primary literature. Core biochemical laboratory techniques such as tissue extraction, cell lysis, centrifugation, and purification were practiced, along with quantitative methods for confirming experimental yield. Following the completion of the project, our group designed a scientific poster in addition to this report.
The poster can be viewed at the following link:
https://www.slideshare.net/MichaelDeBrota/agarwal-a-debrota-m-fenimore-l-purification-and-quantification-of-cytochrome-c-from-bovine-liver-tissue-project-poster
Michael DeBrota et al. - Assessment of Computational Histopathology in Thorac...Michael DeBrota
Background and Hypothesis:
Thoracic aortic aneurysm (TAA) histopathology includes elastic fiber (EF) abnormalities, mucoid extracellular matrix (MECM) accumulation, and smooth muscle derangement in the aortic medial layer. While semi-quantitative grading of these characteristics is a standard practice, computational characterization of medial layer components may facilitate novel quantitative analyses at higher throughput. We hypothesized that computational results would correlate with results of semi-quantitative grading of aortic histopathology.
Experimental Design:
Formalin-fixed, paraffin-embedded human aortic tissue sections were stained with Movat’s pentachrome to characterize aortic microstructure. Sections were also immunostained for nitrotyrosine residues to assess oxidative stress. Samples were initially graded semi-quantitatively by two independent blinded readers. Next, computational histopathology software was used a) to quantify the proportions of EF, MECM, and cellular area in the medial layer of pentachrome-stained sections and b) to quantify the distribution and intensity of positive nitrotyrosine staining in immunostained sections. Association between semi-quantitative grading and computed values was tested with ANOVA.
Results:
The cohort included 74 participants who underwent prophylactic aortic replacement for TAA and 23 healthy controls. The mean age was 54±17 years. On average, EFs accounted for 49% (range 6-90%) of medial tissue area, whereas MECM accounted for 25% (1-73%). The overall semi-quantitative grade of medial degeneration severity was associated with decrease in EF fraction (p=0.02). The grade of EF thinning also strongly correlated with decrease in EF fraction (p=1x10-6). Meanwhile, grade for accumulation of MECM was associated with increase in MECM (p=0.004). Increased semi-quantitative grading for nitrotyrosine levels was associated with increased nuclear signal optical density (p=9x10-10) and greater percentage of cells labeled as strongly positive (p=8x10-10).
Conclusion and Potential Impact:
We observed significant correlations between computed quantitative values and semi-quantitative grading. This suggests that computational histopathology is a valid method for investigation of human TAA tissues.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
In silico drugs analogue design: novobiocin analogues.pptx
Beasley J, Craig B, DeBrota M, McClure W - Isolation and Classification of Soil Microorganisms with Suspected Antimicrobial Properties - Project Poster
1. Isolation and Classification of Soil Microorganisms with Suspected Antimicrobial Properties
Jon-Michael Beasley, Bailey Craig, Michael DeBrota, Jake McClure
Introduction
Materials and Methods
Results Conclusions
References
Rose-Hulman Institute of Technology, Terre Haute, Indiana
Antibiotic resistance is an increasing problem in the areas of public
health and pharmaceuticals. To combat this resistance, research is
constantly being conducted in an attempt to discover novel antibiotic
compounds. Some naturally-occurring substances, including
microorganisms, have been found to exhibit antimicrobial properties. In
our experiment, closely modeled after the protocols of the Small World
Initiative: Research Guide to Microbial and Molecular Diversity,
microorganisms were isolated from soil samples taken on the Rose-
Hulman Institute of Technology campus, tested for antimicrobial
properties, and classified.
Materials
● 10% TSA agar plates
● Inoculation loop
● Bunsen burner
● PCR Thermocycler
● Microcentrifuge
● Gram staining materials
● Stereomicroscope
● Biochemical testing materials
Methods
Five different soil samples were collected from various locations on the
Rose-Hulman campus. These soil samples were serially diluted and
plated on 10% TSA plates. For each soil sample, 32 colonies were
selected from the first countable serial dilution plate and grown on a
patch plate. A new patch plate was created from the original, and a top
agar inoculated with a safe ESKAPE pathogen relative was poured
over the new patch plate. The new patch plates were then incubated
and inspected for zones of growth inhibition of the ESKAPE pathogen
relative, indicating which colonies exhibited antibiotic properties. Four
of the antibiotic-producing isolates were chosen for classification. PCR
was performed to amplify the 16S rRNA gene and gel electrophoresis to
confirm results. The isolates were classified by Gram staining,
microscopy, biochemical tests, and sequencing of the 16S rRNA gene.
Isolate 1 2 3 4
Gram Stain + - + -
Hemolytic Activity (Sheep’s
Blood)
- +Beta
Triple Sugar Iron Agar
Slants
- +(all)
MSA Media + +
Catalase + +
Oxidase + - - -
Motility - - + -
Simmons’ Citrate - -
EMB Media - -
Indole - - -
MacConkey’s Media - -
Isolate Bacterial Morphology
1 Circular, Flat
2 Coccobacillus
3 Bacilli, Central spore
4 Filamentous
Figure 4: The bacterial morphology was
determined by examining each isolate.
http://www.tgw1916.net/bacteria_logare_desktop.html
http://www.microrao.com/identify.htm
Holt, John G., and David Hendricks Bergey. Bergey’s
Manual of Determinative Bacteriology. Williams &
Wilkins, 1994.
“Small World Initiative: Research Protocols and
Research Guide to Microbial and Chemical Diversity
Package.” XanEdu, Small World Initiative,
www.xanedu.com/higher-
education/educators/custom-books-catalog/small-
world-initiative-research-protocols-and-research-
guide-to-microbial-and-chemical-diversity/.
Figure 1: Gel elctrophoresis
reveals PCR was successful for
isolates 1 and 2.
Isolate Identification
1 Viridibacillus neidei
2 Acinetobacter baumannii
3 Bacillus cereus
4 Shigella sonnei
Figure 2: Isolates 1 and 3 (first and third images) were confirmed to be gram-
positive after a successful Gram stain, indicated by purple coloration. Isolates 2
and 4 (second and last images) were confirmed to be gram-negative, indicated by
pink coloration.
Figure 3: Table of biochemical test results for each soil isolate.
A combination of Gram
staining, viewing the bacteria
under a microscope, and
biochemical testing were useful
in classifying the antibiotic-
producing isolates.
The following are our final conclusions
for the antibiotic-producing species
contained in the soil samples:
Unfortunately, our PCR sequencing did
not yield results that supported the
other tests. A form of Pseudomonas took
over many of the plates, including
isolates 1 and 2, and the PCR
sequencing was unsuccessful for isolates
3 and 4.
Figure 5: FinchTV nucleotide sequence for
Isolate 2.
Location CFUs/g soil # Antibiotic-producers
Speed Lake 450,000 4
Hill, Mees/Union 520,000 6
SRC Creek 460,000 13
Base of tree, Scum Pond
location
310,000 1