1. Mohammed Kazzeh RDC S00127084
Project Title:
Experimenting the theoretical potential of a novel seaweed extract for controlling
Pseudomonas infections.
Layman-friendly summary:
These Pseudomonas infections are typically initiated by a common bacteria called
Pseudomonas aeruginosa. It is usually carried by healthy people, this bacteria creates
microbes which cause difficulties such as acute external otitis and hot tub folliculitis. This
infection is the best predominant pathogen of cystic fibrosis infection of the lungs. If a
person is ill or weak these microbes can cause severe difficulties to the body. This disease is
tough to treat as the microorganisms can repel several kinds of antibiotics. (WebMD, 2015)
The aim of this research grant proposal is to find a way to treat the infection for the
minority of people which are weak, ill or healthy. Best method is to try and prevent by
general hygiene and vaccines. If that doesn’t work than normally antibiotics such as
gentamicin are used to cure but for immuno-comprised patients it is sometimes relatively
difficult to cure. (Bonsall, 2015) In my research I plan to eradicate the problem of developing
Pseudomonas infections by experimenting the theoretical potential of a novel seaweed
extract. Seaweed extract from marine macro algae show that they have potential
pharmaceutical and health promoting benefits as they are a rich source of bio-chemicals.
Scientific summary:
Pseudomonas aeruginosa are gram negative which are normally found in soil, water, plants
and animals. It is a major problem that causes acute and chronic diseases in people that
have cooperated defences in their body. It has been found that these infections occur in
biofilms which meet oxygen constraint.(Filiatrault, 2006) These infections are becoming
hard to treat because they have constitutive manifestation of AmpC β-lactamase and active
efflux mechanism, joined with a small absorptivity of the outward membrane. It has
extraordinary talent to obtain advanced confrontation contrivances to double groups of
anti-microbial agents, comprising of β-lactams, aminoglycosides and fluoroquinolones.
(Strateva, 2009) The composites which existed to devise antiseptic movement were
organised in the method of a colloidal interruption for oral management were GC-MS. The
extracts mentioned could be used as prophylactics. The use of algal excerpts for the
management of Pseudomonas infections is a cheap operational and ecological pleasant
method. (Thanigaivel, 2015)
Introduction:
Pseudomonas infections can reason infections to individuals and are typically originated in
water and on plants. There are some Pseudomonas infections which are not plain and can
be simply cured such as dermatitis. But in this research we look at the infections which are
severe. Most of these infections typically happen in patients which are previously distress
from a severe infection. Pseudomonas can also cause healthcare illness in hospitals which
can infect patients in the hospital. A patient can defend themselves from receiving a
Pseudomonas infections by washing with soap and drying ears after swimming, and asking
2. Mohammed Kazzeh RDC S00127084
your doctor for assistance to treat the infection. Therefore we experiment with seaweed as
its biomass has high value and is already being used in food products. When the bioactive
complexes in seaweed were researched we found that there were nine species. (Løvstad,
2011) The products of the ocean provide a wide collection of natural products. There are a
number of successful pharmaceuticals such as dolastatin 10. This products have been used
for cancer chemotherapy, drug resistance and a number of emerging infectious diseases.
The use of novel seaweed extract has so much benefits with many of the polysaccharides
having indicative bioactivities of high pharmaceutical value.(M D. , 2003) From researching
about seaweed a fucoidan or laminaran is to be removed from a brown alga called
Ascophyllum nodosum, this was to be done by an approach called innovative low-chemical
procedure. These bifidobacteria are live without air, split sugar, great GC comprise, gram-
positive bacteria which are incapably of moving and do not form spores or supply gas by
metabolism. Research has been done on how to isolate, and the enzymatic modification and
bioactivity of algal polysaccharides from the brown macro algae. A number of probiotics are
already being used like galacto-oligosaccharides in food, especially this probiotic which can
moderate the microbiota and encourage bifidobacterial development in an infant and adults
intestinal tract. It was accepted that not all B. breve strains own endogalactanase movement
also by performing relative genome hybridization it was tranquil to institute the
manifestation of endogalactanase movement anywhere it is severely related by the
existence of the galA gene in the Bifidobacterial breve strains.(Motherway, 2013) Portions
which are augmented in exact polysaccharides have recognised positive effects on the
development of Lactobacilli and Bifidobacteria in vitro. The movement of exact portions has
been improved through selective alteration with novel fungal enzymes and a freshly
determined scientific test/dietary interference study has established in vivo assistances.
Many plant advances influence turn out to be advances of the streptophte algae and other
algae cell wall structures might devise even profounder origins and share backgrounds with
further historical algae descendants. Additional classification of algae hey wall
polysaccharides and the enzymes that manufacture them might expose the presence of
essential structures mutual to eukaryotic cell walls. (Popper, 2010) Current assembly
investigation has established that an algae polysaccharide rich excerpt is manufactured by
an organised low chemical technique. This has a noteworthy development despondent
adaptable properties in vitro in contradiction of explicit organisms such as Pseudomonas sp.
that confront patients in a clinical location. The structure of the excerpt has existed to be
resolute in feature in a struggle to compare assembly with natural purpose. The excerpt
which is experiencing preparation and toxicology examinations to test its appropriateness as
a nutritional element. Although, any upcoming requests as a possible bacterial regulator
mediator for relevant or nutritional submission in a scientific location necessitates a
comprehensive understanding of its strength in contradiction of a variety of clinically-
relevant strains and strain flexibility. (Domozych, 2012) This infection is a very common
nosocomial pathogen responsible for significant morbidity and mortality worldwide. People
can get in contact with this disease by exposure to a contaminated source within hospital
environment.
3. Mohammed Kazzeh RDC S00127084
What is the aim of my research on this topic?
The main working hypothesis aim for this project is to carry out an investigation on the
novel algal extract which has great potential for the control of the growth of Pseudomonas
sp. including clinically-relevant strains, particularly in exterior ear infections. If these
infections are not treated it will not stop on its own but it will increase if there is no action
or treatment done. There are treatments such as antibiotics, creams for rashes and pain for
minor problems of this infection. This project will propose to test the hypothesis by
screening the development reply of a panel of Pseudomonas strains using a current micro-
assay system. My aim is to research this novel seaweed extract and to try and eliminate this
infection in Ireland and the world.
Why did I choose to do my research on this topic?
This investigation effort will demonstrate innovation and important chances for novelty.
This study will make the student reason and query the consequences and results established
and also permits the student to contribute in group meetings. This research will syndicate
the subjects of microbiology and biochemistry which will be brilliant for this research. I find
glycobiotechnology to be fascinating in its submission to recognising and emerging novel
polysaccharides and oligosaccharides that control microbial development. I revealed that
glycobiology is developing fast as a principal ground of attention for bio-molecular and
biomedical investigation. To review biochemical and microbiological structures of growth
response of Pseudomonas sp. containing the clinically-relevant strains.
Methodology:
This experiment was to be completed approximately in 7-8 weeks.
1. The microbial cell culture was to be done using petri dishes of agar-based growth
medium. The desired bacteria is added to the petri dishes to inoculate using aseptic
techniques.
2. End-point analyses was to be done with the application of the first culture batches
on the target test extract and control which was glucose.
3. Microscopic analysis of cultures was to be done by determining the cell
morphologies and features and any changes to be recorded.
4. DNA extraction by purification of DNA from the sample by the amalgamation of
physical and chemical approaches. Amplification enumeration of bacterial cell
numbers by qPCR (quantitative polymerase chain reaction) for nucleic acid
quantification.
5. PCR product sequencing using the right fragment and also removing all residual PCR
primers and independent nucleotides. The bioinformatics analysis was done using
proprietary software.
6. HPAEC (high-performance anon-exchange chromatography) and wet chemistry
analysis of carbohydrates and examination of resolvable mono-saccharides and
oligo-saccharides in culture broths to define bacterial exploitation and possible
breakdown of the excerpt.
7. Examination of possible microbial adaptableness to the excerpt in micro-plate and
chemo-stat cultures. The samples will be taken at scheduled intermissions,
4. Mohammed Kazzeh RDC S00127084
administered end-point breakdown will include cell totals. The quantification of
possible bacteriostatic or bactericidal properties and qPCR constructed gene specific
marker assays. The samples will also be treated and warehoused for upcoming
proteomics and marker assays.
8. The transcriptomics analysis by description of most of the transcriptional movement
and or by choice subsection of RNA transcripts,
9. Preparation of reports and posters, and presentation for the research team to show
their discovery.
10. If the results are found to be reasonable, clinical trials would be carried out on this
research.
Gantt chart:
Phases Tasks Weeks
W1-3 W3-5 W5 W5-7 W8
1 Microbial Cell Culture
End-Point Analyses
Microscopic Analysis of
Cultures
2 DNA Extraction
Bioinformatics Analysis
3 HPAEC Analysis
4 Investigation of
Microbial Adaptability
Transcriptomics
Analysis
5 Data Check and Thesis
Preparation
Conferences
Publication
5. Mohammed Kazzeh RDC S00127084
ExpectedOutcomes:
Once completed the pupil will examine quantity reactions to the algae excerpt, possible
bacteriostatic or bactericidal properties by alive/deceased examination as well as possible
deprivation or consumption of the polysaccharide-rich excerpt by HPLC/wet chemistry
assays by the bacteria. Microbial cell development will be observed by nephelometry at
600nm by using Tecan GENios systemwith listing of bacterial cell quantities by plate counts
and qPCR. Microscopy will be used to examine morphological properties on microorganisms
and colony/biofilm creation also for bacterial imaging. One control and one scientific strain
will be examined in micro-plate and chemo-stat cultures to acquire original results on
possible strain compliance. This effort will purpose to offer a firstly, significant vision into
the possibility of the algal polysaccharide as an original, harmless biomaterial for the
regulator of the development of scientifically appropriate Pseudomonas sp. The pupil will
acquire measureable information to provision downstream growth and usage of the algal
excerpt in scientific submissions such as regulating of the external ear infections. This
research when completed will give other scientists the opportunity to carry out other
investigations and experiments relating to this subject and to carry out clinical trials to see
whether it works or not. If it is found to work perfectly and have no side effects it would
cost a lot of money to sell and glycobiotechnology would be researched in further detail
because of its biomedical opportunities. Also once put onto the marketplace, this infection
management will have additional duplicates and many dissimilar management categories.
Budget:
Staff (€)
PhD student tuition fee 1,750
PhD student capitation charge 850
PhD maintenance grant 11,000
Equipment (€)
Dionex HPAEC-PAD system 1,000
Temperature controlled micro-plate reader 5,395
Thermo cycler 155
DNA and Protein Electrophoresis
equipment
225
Microscope 150
Materials (€)
Light Cycler 480 Multi well 96 plates (white)
for gPCR
364
Chemicals 5,500
Travel (€)
2 conferences 3,500
6. Mohammed Kazzeh RDC S00127084
Item Total Cost
Staff 13,600
Equipment 6,925
Materials 5,864
Travel 3,500
Total 29,889
Plans for Dissemination:
Once done all the informationwillbe assembledtogetherandwill be accessibleapplicablyinthe
thesis.Once the thesisisdone itwill be inspectedandcheckedbyreliablepeers.If there are any
identifiable errorsestablishedbythem,the thesiscanthenbe amendedif required.The audiences
will thenlookatthe scientificandlaymanof the thesistounderstandthe researchgrantproposal.
For this researchproposal totake place there has to be two dissimilarkindsof conferenceswhere it
will holdtwodissimilarkindsof assemblies.WhenpresentingusingPowerPointPresentationthe
presentationslideshave tohave applicablelanguage andlayouttopermitthe viewersto
comprehendwellaboutthe investigationandthe outcomesandresultsattainedinthe research.The
firstconference willallow the viewersto comprehend the scientificportion whichwill allowthemto
comprehend whathappened inthe experimentandthe results.Thiswillallow the viewerstocarry
out theirownexperimentonthisresearchonce theyhave abetterunderstanding. Thiswillalso
allowthemtocarry outclinical trialsonpatientsonce the treatmenthasbeenfoundtowork
perfectlyandhave noflaw.The secondconference will allow the wholeworldnotjustthe viewersto
comprehendthe reputationof thisresearchandunderstandthe researchdone.Once the viewers
have understoodthe researchtheywill be able tocarry out new thoughtsandmore assetintothis
area of glycobiotechnologyforevenmore investmentandforfurthermore researchinthe coming
future. Once all of thisis complete itwill be placedonscientificjournals,newspapers,articlesand
websites.
References
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analysisof individual datafrom61 prospective studieswith55 000 vasculardeaths. The
Lancet.
Filiatrault,M.J.(2006). Identificationof PseudomonasaeruginosaGenesInvolvedinVirulence and
AnaerobicGrowth. American Society forMicrobiology.
Goodwin,C.D. (1976). Improvedmethodsforthe studyof hepatic. Journalof Lipid Research.
7. Mohammed Kazzeh RDC S00127084
HPSC.ie.(2015, 07 22). RetrievedfromHealthProtectionSurveillance Centre:http://www.hpsc.ie/A-
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M, D. (2003). Marine natural productsand theirpotential applicationsasanti-infectiveagents.
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NCBI.(2015, MAY 4).RetrievedfromNCBI:
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Popper,Z.A. (2010). Beyondthe Green:Understandingthe EvolutionaryPuzzle of PlantandAlgal
Cell Walls. PlantPhsiology American Society of PlantBiologists,373-383.
Stary,H. C.(1995). A Definitionof AdvancedTypesof AtheroscleroticLesionsandaHistological
Classificationof Atherosclerosis. Circulation.
Strateva,T. (2009). Pseudomonasaeruginosa –a phenomenonof bacterial resistance. Journalof
Medical Mmicrobiology.
Thanigaivel,S.(2015).Differential solventextractionof twoseaweedsandtheirefficacyin
controllingAeromonassalmonicidainfectioninOreochromismossambicus:A novel
therapeuticapproach. Elsevier,56-64.
webmd.(2015, 05 11). Retrievedfromwedmd:http://www.webmd.com/heart-disease/what-is-
atherosclerosis
WebMD.(2015, 07 20). Retrievedfromhttp://www.webmd.com/a-to-z-guides/pseudomonas-
infection-topic-overview
