Category: Conversation

• General Purpose: To persuade the audience to take immediate action.
• Specific Purpose: To persuade the audience to take immediate action and donate to the Metropolitan Museum of Art.
• Central Idea: The Metropolitan Museum of Art is a nonprofit organization worthy of donation because of the conservation work it provides, the enrichment attendees get, and the inspiration it provides to its visitors.

Introduction – Step #1 – Attention

• Attention getter: I remember the very first time that I spoke in front of this class, I opened up my speech with this quote from my favorite artist William Haynes. “Life, if you do it correctly, is a piece of art. The best part of art is when you finish it and look at the painting. That’s why death is ok.”
• Reveal the Topic: The reason that I’m bringing this quote up is not to reminisce about the beginning of these last sixteen weeks and think about the fact that our time together is nearing the end, it is to bring up what is at the core of this nonprofit organization: Art. To be more specific, this nonprofit is an art museum that is regularly placed as one of the top ten art museums in the world, it is the Metropolitan Museum of Art.
• Credibility Material: What makes me qualified to ask you to donate to this museum is that I am an emissary of artistic expressionism and the efforts to conserve it, and totally not just some guy in a speech class trying to fulfill the requirements of a persuasive speech assignment because… That doesn’t sound nearly as cool or fancy. On a serious note though, to erase any of the scruples you may have about why I am trying to sell you on this museum, it’s simply because I enjoy art.
• Preview Statement: I’m here to tell you all a little bit about what the Met brings to the table as a non-profit, and hopefully move you to make a donation.

Main Point I and Sub Points – Step #2 – Need

Main Point I: The Met brings value to society that can be overlooked.

Sub Point A: The Metropolitan Museum of art has to offer something to the world, it had 7.4 million visitors in 2018 alone they must get something from the visit, so what did they get out of it…an enriching experience. There are not many places one can go to view artwork, you can see it online whenever you want but that feels less concrete, so the museum offers an experience that can’t be attained from a screen. Whether it is a guided tour or aimless wandering around looking at pieces in the museum, you can witness the beauty of art.

Sub Point B: The Met also offers an educational aspect that has been lacking in recent years in some school systems: art classes. Budget cuts and a focus on subjects that students get tested on have left art programs on the back burner for a number of schools, as a result, some schools offer fewer art classes than they would have in the past, and there are even some schools that offer none at all. That is where the Met comes in, it offers an extensive amount of art classes, and these eight pictured here are just the tip of the iceberg.

Connective: The Met offers an artistic experience to visitors, art classes to children, and even classes for adults, though it still holds more weight than I let on, and that weight comes from the effect it has on people and the important work it does.

Main Point II and Sub Points – Step #3 – Satisfaction

Main Point II: The Met’s great importance goes beyond holding art classes, and housing art.

Sub Point A: Art itself is just the expression of creativity fueled by emotions and ideas. The Met houses this art, but it is much more than the glorified storage building some people might think it is. Inspire More Stories is located on the museum’s website, telling the stories of some of the people touched by the Met, and aside from these three people here, there are six more pictures of people who were moved and inspired in some way by the art contained in the Met, but these nine lives impacted by the Met are just a drop in the bucket when it comes to the people inspired from the art; all these individuals who were inspired have the potential to inspire others, whether it be through their professional lives, or in their interpersonal relationships. This inspiration that occurs wouldn’t be possible without intact pieces of art available for viewing.

Sub Point B: Take a look at this copper figure, it is around five thousand years old, dated as being created Circa 3000 B.C.; now think about the fact that there are more than two million pieces of art contained in the collections of the Met’s three locations. That is a lot of art that needs to be maintained because not all pieces of art are as sturdy as a copper alloy sculpture so they may have crumbled a little among the sands of time, and that isn’t taking into account all the accidental or purposeful damage caused by, let’s call them energetic museum-goers. The cost of the processes to restore and maintain these works, as well as the equipment involved can be quite expensive, but without a way to fund this maintenance, the pieces that sparked the inspiration that roped in the hearts of many to the art world would eventually be beyond repair, the Met would become a failure, and that immense inspiration would fizzle out and die…wow that got kind of dark.

Connective: Luckily the Met seems to not be a house of broken dreams just yet, and you can keep it from becoming one by donating.

Main Point III and Sub Points – Step #4 -Visualization

Main Point III: Picture what a contribution from you to the Met could mean.

Sub Point A: A donation from any one of us will ease the burden on the shoulders of a museum trying to stay afloat. The Met may seem like an extravagant museum, but for the past few years, the Met has ended its financial year with millions of dollars worth of deficit. The report from this slide details how the Met had a projected deficit of $23 million with an $8 million dollar deficit budget, so they would have been in the red by $15 million dollars. In actuality they ended 2016 with a deficit of $10 million dollars, $2 million over budget sounds like a blessing compared to 15, but consider the fact that these losses have stacked up over the years to result in tens of millions in losses. These losses threw a wrench in the museum’s plans for the future. With a donation from you, your funds can alleviate that debt, and be put to good use. These donations can go to more arts programs, filling the positions of conservators and other staff that was laid off because of the debt, and going to the creation of new art wings.

Sub Point B: You may be wondering how your donations could do that, even if the entire class chipped in whatever they could part with it wouldn’t be enough to achieve any of the things I just listed; but it would be more than enough to assist the cause. The reason that it would be more than enough is that quantity is the catalyst for qualitative change. Just imagine if every single one of the 29,000 students that were enrolled at UMSL in 2018, donated a dollar, that’s already a large amount of money. It doesn’t matter if you donate one dollar while someone else donates one million dollars, each contribution matters, and they matter equally because they build off of each other to become larger than what they were in the beginning. Just think the donation you make may very well inspire the next generation of artists.

Conclusion – Step # 5 – Action

Signal the End: I’m almost out of time, so before I go let me tell you how to donate, and let me tell you the wondrous benefits you get when you donate because everybody loves benefits. If you give a donation of any size to the Metropolitan Museum of Art, you get a free ticket to the Met Gala—wait…..no, this note says, you don’t get a free ticket to the Met Gala, but what you get instead is just as good. If you donate any amount to the Met, that donation is 100% tax deductible, within the parameters of the law, and that coupled with the ego high you’ll be riding for a month afterward is just as good as free Met Gala tickets. Right?

Reinforce the Central Idea: Moving on there are two different sets of steps to donate on the Met’s website, depending on the size of your screen. The first is to simply hit the make a donation option on the top right area of the homepage, and you are brought to the donation page. The second way to donate is to go to the Met’s home page, click on these three stacked bars in the top right corner, click on join and give, click on support the Met, scroll down to the button that says donate to the Met, and after you click on it just follow the instructions and your donation is complete. This will help the Met continue to fulfill the duties that it set out to accomplish.

As per the popular travel website Trip Advisor, the visitor experience of Machu Picchu is considered a once in a lifetime, unmissable experience, being described as amazing and mystical. Due to the nature of the area, there is a reasonable amount of hiking to be done at the site in order to get the most out of the visit. A popular option employed by the more adventurous tourists is to access the citadel through the Inca trail. The visitor sessions are divided into two sections, the morning session and the afternoon session, covering four hours each. Visitors are seen to have the best experience early morning, where the cloud cover is less and the sunrise presents for an incredible view. It is also advised to take a tour guide, who can give a lot of context and information about the heritage of the site, as well as the relevance of the various structures and ruins present at the site. Many guides have been praised for their insights of Machu Picchu, with some reviews calling the hiring of a guide a ‘crucial’ part of the experience. With its scenic location and rich biodiversity, it is no wonder that many visitors find many panoramic views around this mountaintop citadel. Staying at the nearby town of Agua Calientes is a good idea for tourists who want to explore the entire site.

While the majority of the reviews and experiences of visiting Machu Picchu are positive, there are quite few areas where the visitor experience can be improved. A common theme that we can find from the negative reviews of the site is the problem of overtourism, with the constant sight of people and buses ruining the aesthetic and overall experience of the place. Reviews have also stated that these cause long lines and wait times, which tend to dampen the experience further, with the worst time being the high season in July. Another issue faced by visitors is the lack of prepared guides, with one review stating that the guides were unprepared and provided inaccurate and varying stories about the sites, and another stating that the guide did not tell them anything that they could not have found in a guidebook. Another review noted the lack of safety barriers and proper signage, which is incredibly dangerous given the location of the sanctuary.

To resolve the issues faced by current visitors, we can look at the recommendations made by Pedersen in his 2002 Manual regarding World Heritage Management. He recommends employing the use of tour guides to monitor the tourist behavior at the site. I believe that this can also be applied to ensure tourist safety, thus eliminating the need for safety barriers which could potentially cause damage to the site. This could also work in the benefit of the local community, generating more employment to match the size of the crowds at the site. The number of tour guides should be proportional to the tour groups as well, and should be included in the current number of people allowed at the site. Financing of training of guides can come from tour operators, which can go a long way in enhancing the overall experience. Visitor data acquired by these operators can be used to form demographics on tourists arriving, such as languages spoken by the visitors as well as their motivation behind visiting Machu Picchu, which can be useful in the training of the guides to result in better interpretation of the area by the tourists. Education programs detailing the significance of the area, its resources and the issues that threaten can sensitize tourists about the area and aid in developing respect for this historic and natural marvel before they enter. Pedersen also recommends restricting tourists to parts of the site that have higher durability in order to protect the site. These areas can also be strengthened during the routine maintenance that occurs during the month of January. The inaccessible sites can be experienced through audio-visual aids, guidebooks and through the expert knowledge of the certified tour guides. An increase in visitor charges and additional facilities can be useful in deterring the number of tourists that arrive while supplementing the loss of revenue from this reduction. As per the findings of Larson and Poudyal in their 2011 analysis of Machu Picchu, local development and Peruvian culture were threatened by tourism profits going to the operator agencies. They also found that Andean people viewed international tourism as a threat to their culture and sacred sites. Ensuring that tour guides are from the local community can increase local benefits, deter any increase in tourist facilities that might cause harm to the site, and support a genuine cultural exchange and understanding between the global community and that of the Andean culture, the direct descendants of the Inca civilization responsible for the building of the historic sanctuary of Machu Picchu. The Peruvian government and the Tourism Board can also develop eco-tourism programs that can aid in the conservation of Machu Picchu, and redesign the tourism policy of Peru to redirect the large tourist numbers to other parts of Peru, such as the ruins of Choquequirao, taking the pressure of Machu Picchu.

Organisms are not as it were the adversaries, but they creating income is an approach to form them self-sufficient. Practices just like the fabricate of biogas or conventional microbial matured nourishment items like pickles, vinegar, purpose, and kumis may offer assistance the poor people in producing a source of salary by learning the correct abilities required for deciphering these strategies into an genuine commerce. Microorganisms, even though their small size, play an vital part within the evaluation and support of nourishment quality and security in nourishment handling and fabricating businesses. Their financial imponance which incorporates pharmaceutical, clinical, Nourishment deficiencies, generation of bioenergy, fossil fuel deficiency, soil richness, for producing clean power etc.

Automating our generation forms has made a difference us make extraordinary improvement, but has too caused natural contamination. In any case, we are on the starting of cheerful advancements towards more feasible forms, in portion due to the utilize of microorganisms. Biotechnology is any procedure which includes the application of natural living beings or their components, preparing to fabricating and benefit businesses to form or modify items Biotechnology occupies a really vital position within the socio-economic headway and advancement of the country in specific and the world at expansive. its gives data around biotechnology with illustrations of how these modern apparatuses of science and farming are utilized in nourishment generation as portion of financial progression and national advancement. It moreover incorporates a extend of advances utilized to handle the crude nourishment materials created by the edit, fishery and animals sectors.

Probiotic organisms are known to extend craving and longings for particular nourishments, in this way making a difference overcome. nourishment preparing too produces income, being one of the foremost imperative sources of business and job for the populaces of Africa, Latin America, and Asia. Nourishment things like matured cereal items, aged drain items and matured alcoholic drinks are a critical supporter to their economies.

Utilize of natural forms or operators such as certain microorganism to change over natural materials, such as plant or creature squander, into usable items or vitality sources known as bioconversion forms or biotransformation. Microscopic organisms, actinomycetes, yeasts, and molds have been utilized in different bioconversions.

Organisms can too be utilized for creating clean power. For occurrence, Shewanellaoneidensis and Geobactersulfurreducens can utilize natural matter such as starch or sugar and deliver utilizable power from it. This appears the cutting edge potential of organisms for creating greener and cleaner power. In any case, this energy-saving innovation requires investigate and improvement for coming into hone.

Biofuels done right can demonstrate to be a significant source of vitality that can be delivered in noteworthy amounts and relieve the worldwide fuel issues. Renewable oil generation at a rate of 3200 GJ ha-1 yr-1 has been found to be attainable from the photosynthetic organism Haematococcuspluvialis, appearing the practicability of supplanting “energy crops” by productive strains of microorganisms This would inevitably moderate carbon dioxide outflows whereas demonstrating to an indeed greener technology.

Organisms can too be utilized for the generation of bioenergy by using squander items like sewage slime, metropolitan strong squander and agrarian squander, having microorganisms like Trichoderma, Aspergillus, Penicillium, and Clostridium demonstrated to be exceedingly productive, to bioenergy generation, the accomplishment of maintainable advancement too requires productivity enhancement, vitality sparing approaches and the presentation of the unused advances which can be executed for its use.

the utilize of green growth as a source of biofuel generation offers an amazing arrangement to these issues. Green growth can develop in a wide number of oceanic situations and have the capacity construct up’>to construct up tall levels of oil, in their biomass. Botryococcus can store up to 50% of the biomass within the frame of long-chain hydrocarbons. Green growth multiply and are simple to control hereditarily. A further advantage is the synchronous era of common co-products. The foremost well known green growth are Chlorella, Navicula, Pavlova, and Isochrysis. The major challenges within the utilization of green growth as a substrate for biofuel generation are their collect, activity, and down spilling.

Microorganisms which might be utilized for mechanical generation should meet certain requirements. The maximum imperative resources of microbial societies for make use of in mechanical microbiology were characteristic substances such as soil tests, waters, and ruined bread and fruit

• The living being ought to be able to develop energetically and quickly within the utilized medium.
• The conclusion objects ought to no longer incorporate dangerous and other unwanted materials, mainly at the off threat that those conclusion items are for inner utilization
• The living being ought to have a sensible hereditary, and thus physiological steadiness.

An living being that transforms effectively is an costly hazard. It may deliver undesired items in the event that an happened transformation remained surreptitiously.

• An living being with ideal efficiency at tall temperatures, moo pH values or which is able to expound agents inhibitory to competitors contains a chosen advantage over others.
• The life form ought to be sensibly safe to predators such as bacteriophages. It ought to subsequently be portion of the basic studiesof an mechanical foundation employing a phage-susceptible living being to endeavor to deliver phage-resistant but tall yielding strains of the organism.
• First and preeminent, the living being must be able of development and item arrangement in large-scale culture.
• It ought to create spores (on the off chance that parasites or yeast) or a few other regenerative cell shape so that it can be effortlessly immunized into the expansive vessels utilized to develop the creating life form on an industrial scale.
• It must too develop quickly and deliver the specified item in a generally brief period of time.
• It must moreover be able to develop in a fluid culture medium reachable in bulk amounts at a moo cost. Numerous mechanical microbiological forms utilize squander carbon from other businesses as major or supplemental fixings for large-scale culture media. These incorporate corn soak alcohol and whey.
• An mechanical microorganism ought to not be pathogenic, particularly to people or financially imperative creatures or plants.

Finally, an mechanical microorganism ought to be agreeable to hereditary control since expanded yields are regularly gotten by implies of transformation and classical hereditary determination methods. A hereditarily steady and effortlessly built microorganism is hence a clear advantage for an mechanical process.

Present day mechanical maturation forms utilized within the nourishment and refreshment industry can be depicted concurring to diverse perspectives. The most common meaning of fermentation is the transformation of a sugar into an natural corrosive or an liquor. However, the term “fermentation” is additionally utilized in a broader sense for the deliberateness utilize of microorganisms such as microscopic organisms, yeast, and organisms to create items valuable to people on an mechanical scale. Any prepare interceded by or including microorganisms in which a item of financial esteem is gotten is called fermentation.

TYPES OF FERMENTATION PROCESSES

Batch fermenter

Batch culture speaks to a closed framework in which the medium, supplements, and inoculum are included to the bioreactor, generally beneath aseptic conditions, at the starting of development that’s , the volume of the culture broth within the bioreactor is hypothetically consistent amid development.

Fed batch cultivation

Fed-batch culture speaks to a semi-open framework in which one or more supplements are aseptically and continuously included to the bioreactor whereas the item is held interior (Figure 4.1b); that’s , the volume of the culture broth within the bioreactor increments inside this time. The most focal points of fed-batch over batch societies are:

• (a) the plausibility to drag out item blend,
• (b) the capacity to attain higher cell densities and in this way increment the sum of the item, which is more often than not corresponding to the concentration of the biomass,
• (c) the capacity to upgrade abdicate or efficiency by controlled consecutive expansion of supplements, and
• (d) the highlight of delayed beneficial development over the “unprofitable periods” when the bioreactor would regularly be arranged for a modern batch.

Continuous fermenter

Continuous culture speaks to an open framework in which supplements are aseptically and persistently included to the bioreactor, and the culture broth (containing cells and metabolites) is expelled at the same time. that’s, the volume of the culture broth is consistent due to a consistent feed-in and feed-out rate. The most focal points of persistent culture (chemostat) over the clump mode are

• (a) the plausibility to set up ideal conditions for greatest and long-term item blend,
• (b) the capacity to attain steady item quality (the unfaltering state is characterized by a homogeneous cell culture spoken to by a consistent concentration of biomass and metabolites), and (c) a unmistakable diminishment in “unprofitable” periods of the bioreactor operation.

Fermentation is an economically-viable and green prepare by which natural products and vegetables can be put away for longer-term utilization. By utilizing organisms like Lactobacillus, Saccharomyces and Acetobacter, a few essential and auxiliary metabolites with wide applications in nourishment conservation are produced. fermentation innovation may be a valuable instrument that can be utilized in several approaches to protecting nourishment, subsequently killing hunger.

Fermentation is all inclusive connected within the conservation of a run of crude rural materials (cereals, roots, tubers, natural product and vegetables, drain, meat, angle etc.). Commercially delivered matured nourishments which are promoted universally incorporate dairy items (cheese, yogurt, and matured milks), frankfurters and soy sauce. Certain microorganisms related with aged nourishments, in specific are strains of the Lactobacillus species, which are probiotics i.e. used as live microbial dietary nutrients or nourishment fixings Nourishment fermentations contribute considerably to nourishment security and food security, especially within the rustic zones of numerous creating nations.

Microbial fermentation products

Production of Curd

Microorganisms like Lactobacillus and other fermicutes bacteria develop in milk, which change over it into curd. We have seen at home that a starter is added to milk which turn it into curd. This starter is known as inoculum, which contains millions of LAB.

During development, LAB create acids that thicken and somewhat process the drain proteins. In this way, changing over drain to curd. These moreover progress wholesome quality by expanding vitamin-B12 substance of the curd. LAB moreover play exceptionally advantageous part in checking illness causing organisms in our stomach.

Dough

It is fermented by bacteria in making foods such as dosa and idli. The puffed up appearance of dough is due to the generation of CO2 amid aging. In bread making, batter is aged utilizing baker’s yeast, i.e., Saccharomyces cerevisiae.

Toddy

It could be a conventional drink of a few parts of Southern India. It is made by yeast aging sap from palms trees, coconut, etc. Organisms are moreover utilized to age angle, soya bean, bamboo shoots, etc.

Cheese

[image: ]It is known to be the most seasoned nourishment thing in which organisms are utilized. It is shaped by halfway debasement of drain by diverse microorganisms. Distinctive assortments of cheese are known by their surface, enhance and taste. Swiss cheese with huge gaps is created by Propionibacteriumsharmanii. Gaps are made due to the generation of huge sum of CO2 delivered by this bacterium. Roquefort cheese is matured by developing a particular organism on them, which deliver them a specific flavor.

Beverages

Yeasts have been utilized from antiquated time for the generation of refreshments like wine, brew, whisky, brandy and rum. Saccharomyces cerevisiae commonly called brewer’s yeast is utilized for bread making, maturing malted cereals and natural product juices to ethanol. Depending upon the sort of crude fabric and forms, distinctive sorts of alcoholic drinks are arranged. Wine and brew are sifted, pasteurized and bottled without assist refining, though whisky, brandy and rum are created by the refining of matured broth. Brew has an alcoholic substance of 3-6%, whereas in wines; the alcoholic substance is around 9-12%

Pharmaceutical

The pharmaceutical companies moreover play a huge part in showcase, items counting architect anti-microbials, having a tremendous showcase request additionally they win over 1-3 billion dollars each year. Coming to agrarian division they too utilize microbial or biotech items counting bio bug sprays, plant development promoters, and bio pesticides. The present day biotechnology industry has made a major affect within the trade world. Biopharmaceuticals (recombinant protein drugs, immunizations and monoclonal antibodies) having a showcase of 15 billion dollars. Recombinant DNA innovation has moreover delivered a insurgency in horticulture and has particularly expanded markets for microbial chemicals. Atomic controls have been included to mutational procedures as implies of expanding titers and yields of microbial forms and in disclosure of modern drugs. Microbial biotechnology has exceptionally tall potential to overcome world economy by commercializing esteem included items to worldwide exchange advertise.

Amino Acids: awesome significance for human utilization and creature nourish enhancement, within the production of aspartame (vital within the delicate drinks industry), make of beauty care products, hyaluronic corrosive utilized in tanning (biotransformation of histidine amino corrosive) etc. • Vitamins: Generation of Riboflavin (Vitamin B2), and cobalamins (Vitamin B12), ascorbic corrosive (vitamin C), β-carotene (provitamin A), biotin (vitamin H) ergosterol (vitamin D2)

Reference

1. The role of biotechnology in the socio-economic advancement and national development: December 2006. https://www.researchgate.net/publication/242675642.
2. The role of microorganisms in achieving the sustainable development goals. (May 2018,). https://www.sciencedirect.com/science/article/pii/S0959652618303871?via%3Dihub.
3. AN INTRODUCTION TO FERMENTATION . Brian Pumphrey and ChristianJulien( May 1996). https://www.wakenbtech.co.jp/wp/wpcontent/uploads/2015/11/nbs_fermentation_basics.pdf

Apurinic and apyrimidind (abasic or AP) sites are the most common DNA lesions that have the ability to block replicative polymerases. Stem cell-specific 5-hydroxymethylcytosine-binding protein (HMCES) covalently crosslinks to the AP site and prevents genome instability. An HMCES DNA-protein crosslink (DPC) with an AP site within a 3′ overhang DNA is able to shield the Ap site from endonucleases and error-prone polymerases. The conserved SOS-response associated peptidase (SRAP) domain of HMCES and its Escherichia coli ortholog YedK are able to mediate lesion recognition by interaction with a single strand- and double-strand-containing DNA structures found in DNA damage sites. AP site is being protected by a stable thiazolidine linkage, which causes HMCES DPC stability.

Human HMCES contains a single domain called SRAP proteins, which is 38.5% identical to that of E. coli YedK. KAREEM 2019 SRAP is a protein made up of an invariant cysteine, a conserved histidine, and glutamic acid and existed inside a putative catalytic pocket. A positively charged surface is available next to this pocket. DNA binding depends on the arginine lined on this surface. KAREEM 2019 Regarding binding to DNA, both HMCES and YedK strongly prefer ssDNA compare to dsDNA ligands. Wang 2019 These DNA-protein crosslink (DPSs) are exposed to DNA damage that generates AP sites. KAREEM 2019AP site is repaired by base excision repair (BER), which is dependent on an intact DNA duplex. Not only base loss happens in ssDNA more rapidly than dsDNA, but also dsDNA AP sites that have not been repaired are converted into ssDNA. AP sites and replicative polymerase will stall at the AP site and leave a 3’ dsDNA-ssDNA junction PETRIA 2019. It has been revealed that AP are typical DNA lesion and lead to hypersensitivity of HMCES cells to DNA damaging agents. KAREEM 2019 DPC formation in cells is dependent on the HMCES catalytic cysteine and DNA binding surface. Therefore, HMCES is known as a sensor of AP sites and covalently modifies the ssDNA AP site to generate a DPC. KAREEM 2019 Here, we discuss a recent discovery of HMCES as a sensor which shield AP sites in ssDNA at replication forks.

Main

DNA glycosylase and glycosylase/abasic (AP) lyases are the enzymes that initiate the base excision repair pathway. In such a process, they recognize the damaged target base and catalyze the breakage of the base−sugar glycosyl bond. DPC can be formed by covalently binding of AP site to PARP-1 during alkylating agent-induced base excision repair (BER). This complex formation depends on existence of natural AP sites in DNA and single-strand DNA incision Rajendra 2014 It has been showed that detecting of HMCES DPC in cells is comfortable,KAREEM 2019, its incubation does not change the percentage of crosslinked protein, and only a small peptide-DNA linkage remains stable after proteolysis of the DPC with proteinase K. PETRIA 2019 Altogether suggest HMCES DPC as a stable chemical linkage and irreversible in physiological conditions. PETRIA 2019

Both the Trp81Glu substitution at dsDNA-interaction site A and Arg4Ala substitution at dsDNA-interaction site B are essential for substrate DNA binding the HMCES SRAP domain with 3′ overhang DNA which contains an AP site. In order to structure a reactive aldehydic form of the AP site which has the ability to crosslink to HMCES, the 3’ overhang to obtain a deoxyuridine (dU) at position 9 of the longer DNA strand was designed. Levon 2019 The AP site is located above Cys 2 which is at the N terminus of the protein. Addition of an N-terminal GST fusion protein inhibits DPC formation. KAREEM 2019In order to crosslinking happens between AP and Cys2 of HEMES, the N- terminal methionine needs to be removed to expose the NH2 of Cys2 and produce a catalytically active form of HMCES. Cys2 and AP react in ring-opened aldehyde to form a thiazolidine DNA-protein crosslink. Levon 2019

Cys2 belongs to a cluster of three conserved residues that includes Glu 105 and His 160 PETRIA 2019 This kind of evolutionary conserved residues cause DNA and protein sides of the thiazolidine linkage to be stable. PETRIA 2019 The electron density is located clearly the AP in the ring-opened form, so HMCES reacts with the aldehyde in the ring-open way. KAREEM 2019 In addition, a continuous density between C3′ and this cysteine side chain has been showed. KAREEM 2019

The anomeric C1′ carbon of the AP site bond covalently to the α-amino nitrogen and the side chain sulfur of Cys2. When AP aldehyde C1′ carbon by Cys 2 α-NH2 generates a Schiff base intermediate by nucleophilic attack, it causes Cys 2 sulfhydryl group attack C1′ and form a thiazolidine ring. PETRIA 2019 YedK DPC formation is diminished by removing thiol from C2A mutant KAREEM 2019

As in the reaction between cysteine and aldehydes, the equilibrium favors thiazolidine wang 2019 compared to Schiff base, no DNA lyase activity resulted from β-elimination of the Schiff base intermediate can be observed. PETRIA 2019, both the C2A and C2S mutant showed DNA lyase activity when incubated with ssDNA containing an AP site, which is reduced when crosslinking happens in the presence of sodium cyanoborohydride (NaBH3CN). NaBH3CN is a reducing agent which causes the Schiff base intermediate stabilization. PETRIA 2019 Therefore, in this mechanism, the Schiff base intermediate is captured as a result of the nucleophilic attack to the cysteine thiol.PETRIA 2019

DNA also interacts with SRAP inside the dsDNA interaction site, which contains Gly3, Arg4, Pro 46, Asp 47, Trp128. This interaction site is evolved to bind duplex DNA, which represents a potential binding site for 5’overhang DNA. Levon 2019 A crystal structure of YedK linked noncovalently to an ssDNA oligomer containing a C3-spacer in place of the AP site and a protein in the non-covalent complex identical to that of DPC. Wang 2019 In this non-covalent complex, the DNA at the 5’ end is positioned identical to that of the DPC structure, while its 3’ end is mobile, which results in a crystal packing difference between the two complexes. Both these structures show that the SRAP domain accommodates dsDNA on the 3’ side of the AP site.PETRIA 2019 On the 3’ side, all nucleobases of the AP site are stacked in a B-DNA conformation and the residues adjacent to the AP site stacks on the surface of the protein against Pro40 and Ile74 which forms a highly conserved ‘shelf’ which stabilize a base pair 3’ to the AP site. Conservations in this region show that binding to AP sites in the context of a 3’-truncated ssDNA-dsDNA junction is an important feature. PETRIA 2019

A 1.6-Å resolution crystal structure of E. coli YedK covalently crosslinked to a heptamer ssDNA containing an AP site showed that a positively charged channel is formed inside the core β-sheet that structures the ssDNA phosphoribosyl backbone along one face of the protein. PETRIA 2019 This backbone is kinked and twisted by 90° at the AP site and cause the nucleobases of each flanking trinucleotide be orthogonal to one another which explains why SRAP disfavors binding to dsDNA. levon 2019 Among the SRAP domains, residues inside the DNA-binding channel are the most highly conserved ones which shows conservation of DNA-binding modality. PETRIA 2019 Both YedK and HMCES prefer to bind to ssDNA and the mutation of conserved amino acid diminishes DNA binding for both proteins. Wang 2019

The crystal structure of SRAP-DNA complex that contains a three-nucleotide overhang at the 3′ end shows that SRAP binds to both 3’end of DNA and blunt-end of another DNA, while both interaction surfaces were highly conserved. SRAP binding to 3’end happens by a hydrophobic shelf created by Tr 81 and Phe92 and interact with the duplex segment of DNA at the ssDNA-dsDNA junction. Levon 2019 Therefore, as DNA polymerase stall at an AP site leaves a 3′-truncated nascent strand with a 5′-overhanging template, SRAP proteins should function at a stalled replication fork. HMCES is efficient at binding and crosslinking to an AP site immediately adjacent to the 3′ ssDNA-dsDNA junction as to ssDNA but not at the presence of dsDNA on the 5’end PETRIA 2019

To conclude, it has been revealed that HMCES shield AP sites from further processing by exonucleases during DNA replication and promote genome stability. Cells detect DNA lesions by using HMCES as sensors. HMCES recognizes AP sites in ssDNA and modifies the lesion to generate a DPC. HEMCES and SRAP-containing proteins crosslink to AP sites at 3’ end and it has substrate preference because of DNA formation happens when polymerases stall at Ap sites. The thiazolidine ring involves the NH2 of Cys2 and acts as a sink for AP sites and prevents strand breaks. This thiazolidine DPC link and SRAP-AP-DNA crosslink cause genome stability and improve organism fitness.

Abstract

Plant seedling classification is crucial for biodiversity conservation. Based on analysis of CNN, we propose a system to classify plant seedlings with minimum classification error. By machine learning algorithm, convolutional neural network have been applied to different datasets. We used training set and test set of images of plant seedlings at various stages of grown. The dataset comprises 12 plant species. The goal is to create a classifier capable of determining a plant’s species from a photo. The experimental results validate that the proposed method effectively classifies plant seedlings which are there in dataset. The training set achieved an accuracy of 93% and test set achieved accuracy of 95%. In future works, we plan to detect disease on identified plant species.

Introduction

Plants remain an important and essential source of food and oxygen for nearly all living organisms on earth. Agriculture is prevailing in some continents like Africa, therefore appropriate automation of the farming procedure would assist in optimizing the crop yield and ensuring the perpetual productivity and sustainability. In accordance with there is a sturdy bond between raised productivity and economic growth. Thus, the application of smart farming techniques in the agricultural sector can empower the development of the economy in many countries. Seedlings quality assessing proved to be a powerful means of prophesying the growth performance and, hence, optimizing the plant production. Seedling classification is the first step to fulfill the seedling quality evaluation.

Furthermore, the invasion of weeds on farmlands leads to decline in the crop yield. Generally, weeds have no valuable beneficial, regarding nutrition, food or medication. However, they grow very quickly as well as they intrusively compete with original crops for space and nutrients. Weeds identification is not an easy process due to the hazy boundaries of the crops, together with the diverse sandy and rocky backgrounds. Thus, there is a need to develop an efficient technique to accurately and certainly detect weeds from beneficial plants.

In order to improve agronomic production and crop quality, farmers should follow precision agriculture.

Precision agriculture is a farm management approach that utilizes information technology and artificial intelligence to guarantee profit maximization, crop yield optimization, and environment preservation. One of the fundamental challenges that face precision agriculture is weed control. Weed control must be achieved earlier as possible after crop germination before weeds begin to compete with crops for nutrition and cause adverse effects. Thus, optimal weed treatment is recommended in the seedling stage. Nevertheless, in this phase, the discrimination between crops and weeds has some limitations;

• a) Inadequate image resolution for distinguishing between exposed soil, crop seedlings and weeds
• b) Resemblance of spectra and appearances between weeds and useful crops in the early stages
• c) Overlapping of the soil background reflectance with the detection process

The plant seedlings dataset contains images of approximately 960 unique plants belonging to 12 species at several growth stages. It comprises annotated RGB images with a physical resolution of roughly 10 pixels per mm.

The application of machine learning techniques for automatic plant seedling classification has become a significant and promising field of research towards improving agriculture outcomes. Deep learning is a specific type of machine learning that has gained substantial interest in various disciplines. The Convolutional Neural Network (CNN) is a deep neural network architecture that is generally used to analyze visual images. Latterly, CNNs have achieved a significant breakthrough in computer vision fields. Additionally, the CNNs proved to have high ability to obtain the efficient features needed for image classification process. Traditional image classification algorithms, handcrafted features are firstly extracted, then a feature selection process is achieved, and finally, a suitable classifier is chosen. However, CNN is proficient in learning various features from images, it covers global and local features, and it uses these features for efficient classification. CNN showed superior performance compared to other image processing techniques. Therefore, in this project, the enforcement of the CNN approach for plant seedling classification is investigated. We are implementing our project using CNN that is Convolutional Neural Network due to its extraordinary features from other existing techniques.

Literature review

The method proposed by Yang Song et al. (2016)[1] aims to do texture image classification with discriminative neural network. Key area of working is CNN-based features to achieve more accurate classification of texture images, discriminative neural network-based feature transformation (NFT) method for better classification. They come up with enhanced classification performance where CNN-based features (FC-CNN and FVCNN) provides better classification than handcrafted features. Research gap is observed there where evaluation of FV descriptors based on other types of local features that are handcrafted can be done.

The method proposed by Heba A. Elnemr (2019)[2] aims to create a Convolutional Neural Network architecture for plant seedling classification. Key area of working is to use CNN to classify plant seedlings in early growth stage and parameters like accuracy, precision are observed. They come up with accuracy of 94.38%. This system can be used to create IoT system for weed control.

The method proposed by KuoFang Chung et al. (2017)[3] aims to do phylogenetic classification of seed plants of Taiwan. Key area of working is to classify plant species from Taiwan on the basis of biological things. They come up with gymnosperms in 5 families and angiosperms in 210 families. Research gap is observed there where they used dataset consisting of plants only from Taiwan.

The method proposed by Yong Wang et al. (2019)[4] aims to do research on image classification model based on deep convolution neural network. Key area of working is to use depth neural network for maximum interval minimum classification error, M3CE, comparison of SVM, KNN, BP and CNN methods. They come up with accuracy of each methods like CNN,SVM,KNN etc. and they turned out as 99.68%,89.41%,81.25% resp. Deep convolutional neural network as a black box feature extraction model.

The method proposed by Samir S. Yadav et al. (2019)[5] aims to use deep convolutional neural network based medical image classification for disease diagnosis. Key area of working is Deep neural network is capable of classifying images, linear support vector machine classifier with local rotation and orientation free features, transfer learning on two convolutional neural network models: Visual Geometry Group i.e., VGG16 and InceptionV3. Transfer learning deals with retaining specific features and improved performance. They come up with conclusion as CNN-based transfer learning is the best method of all three methods. The capsule network is better than the ORB and SVM classifier. In general, CNN based methods are better than traditional methods. Transfer learning on VGG16 provides best results. Research gap is observed there where they can add visualization at output so that we will get visuals of disease.

The method proposed by Yunus Ozen et al. (2018)[6] aims to detect and classify plant leaf disease based on CNN with LVQ algorithm. Key area of working is to detect diseases like bacterial spot, late blight, septoria leaf spot and yellow curved leaf diseases and necessary action is taken, color information is used for plant leaf disease researches. Leaf disease and classification using CN N and LVQ is done here. Improvement in recognition rate in classification process different filters will be done as far as future scope is considered.

The method proposed by Aydin Kayaa et al. (2018)[7] aims to do analysis of transfer learning for deep neural network based plant classification models. Key area of working is to apply different data sets and use transfer learning to provide important benefits in plant classification. They come up with End-to-end CNN with accuracy 70.79%, LDA and SVM average accuracy reads 95% and 96% resp. for all four plants of dataset. This system can be extended to plant disease identification and weed control.

The method proposed by Jayme Garcia Arnal Barbedo (2013)[8] aims to use digital image processing techniques for detecting, quantifying and classifying plant diseases. Key area of working is to use digital image processing techniques to detect, quantify and classify plant diseases. The selected proposals are divided into three classes as detection, severity quantification, and classification. They come up with detection, severity quantification, and classification for various tools like CNN, SVM etc. Improvement in disease identification, provide pesticides/medicines to cure identified disease are the things on which we can work in future.

The method proposed by Shenghui Yang et al. (2019)[9] aims to use classification method of plug seedlings based on transfer learning. Key area of working is to use classification method for plug seedlings based on transfer learning. Extracting and graying the interest region, average accuracy 100% is achieved for 3 specifications. They come up with accuracy of VGG16 and it turned out 95.50%.

The method proposed by Ruji P. Medina et al. (2018)[10] aims to classify plant seedling images using deep learning. Key area of working is to use image transforms like resize, rotate, flip, scaling and histogram equalization in CNN and data augmentation. They come up with 99.74% accuracy for validation and 99.69% accuracy for testing. This system can be extended to classify herbal plant species from different countries.

The method proposed by Yair Weiss et al. (2019)[11] aims to why deep convolutional networks generalize so poorly to small image transformations? Key area of working is small translations or rescaling’s of the input image can drastically change the network’s prediction. ML tools like CNN, data augmentation and antialiasing are used. They come up with CNN learning invariance from the data augmentation procedure. We can achieve more accuracy though image size is small.

Methodologies

The ability to do so effectively can mean better crop yields and better stewardship of the environment. Huge rise in the number of images that are collected on a daily basis. A large database of images is available worldwide across various platforms. In order to utilize these databases, an effective and robust retrieval and search approach is required. The application of ML techniques for automatic plant seedling classification has become a significant and promising field of research towards improving agriculture outcomes. This process became very laborious and ambiguous due to the rapid increase in the number of images and the diversity of the image contents. Hence, a new method i.e. CNN was devised and is very useful for this purpose.

In area of plant seedling classification, various machine learning algorithms are used. In most of the projects, CNN, SVM, KNN, LVQ etc. algorithms are used and classification is done. Each algorithm achieves different accuracy (in percentage) and has pros and cons. CNN that is convolutional neural network is widely used in image classification as it provides high accuracy as compared to other algorithms. SVM that is state vector machine is another algorithm used in image classification but its accuracy is slightly less than that of CNN. KNN that is K-Nearest Neighbors is another algorithm used in image classification. It is easy to implement. LVQ that is Learning Vector Quantization is another algorithm used in image classification. It is basically prototype based supervised classification algorithm.

CNNs for applications that involve images. Why CNNs are more suitable to process images? Pixels in an image correlate to each other. However, nearby pixels correlate stronger and distant pixels don’t influence much. Local features are important: Local Receptive Fields. Affine transformations: e.g. the class of an image doesn’t change with respect to translation. So we can build a feature detector that can look for a particular feature (e.g. an edge) anywhere in the image plane by moving across. A convolutional layer may have several such filters constituting the depth dimension of the layer. CNN Conceptual Understanding – Inputs: the input layer is a linear arrangement of input elements. This is typically represented as an input vector. CNNS operate on a 3d input ‘volume’. Hidden Layers of the architecture. Neural networks use one or more hidden layers with the units typically performing some nonlinear transformations (e.g. tanh). Convolutional networks use: convolutional layers and pooling layers. Local receptive fields and shared weights for a given filter across the complete input. Strides Filters arranged depth wise: small size. Output layer In a traditional NN the output layer (such as Softmax or Logistic) connect to the linear hidden layer in a fully connected pattern CNNS follow a similar pattern where the ‘volume’ represented by the final internal layer is fully connected to the output layer. CNNs operate on images that have a 2d surface and a depth in terms of RGB colors. Hence they are 3 dimensional. The standard neural network architecture has a linear input layer structure. When we discuss CNN architecture, it helps to decouple in our minds the abstraction CNNs deal with and the practical realization of these principles on a linear layer based neural network architectures. The algorithm here performs in three steps and they are as follows-

• a. Preprocessing- crop parts of image, flip image horizontally, adjust contrast and saturation
• b. Splitting data set
• c. Building CNN

Conclusion

CNN is used to identify, scaling, translation and other forms of images. This project is plant seedlings classification. While doing this project we come to know about various concepts like deep learning, machine learning, CNN i.e. convolution neural network etc. The code had been successfully implemented using CNN in Jupyter and Python IDEs. We also come to know about how plant seedlings are classified using various machine learning tools specially CNN. At output, we got one .csv file which comprises of image file and species which we call it as seedlings classification. In future works, we can detect disease on identified plant seedling. We can classify herbal plants. To increase understanding of details of target object, further research is needed. Another dimension is to perform new experiments when more public datasets become available.

References

1. Yang Song, Qing Li, Dagan Feng and Weidong Cai, “Texture image classification with discriminative neural networks”, Research article on computational visual media Vol. 2, No.4, December 2016, 367-377
2. Heba A. Elnemr, “Convolutional neural network architecture for plant seedling classification”, International Journal of Advanced Computer Science and Applications Vol. 10, No. 8, 2019
3. Cheng-Tao Lin, Kuo-Fang Chung, “Phylogenetic classification of seed plants of Taiwan”, Lin and Chung Bot Stud (2017) 58:52, DOI 10.1186/s40529-017-0206-6
4. Mingyuan Xin, Yong Wang, “Research on image classification model based on deep convolution neural network”, EURASIP Journal on Image and Video Processing, 2019
5. Samir S. Yadav, Shivajirao M. Jadhav, “Deep convolutional neural network based medical image classification for disease diagnosis”, Journal of Big Data, 2019
6. Yunus Ozen, Adem Tuncer, Melike Sardogan, “Plant leaf disease detection and classification based on CNN with LVQ algorithm”, in: 3rd International Conference on Computer Science and Engineering, 2018
7. Aydin Kaya, Ali Seydi Keceli, Cagatay Catal, Hamdi Yalin Yalic, Huseyin Temucin, Bedir Tekinrdogan, “Analysis of transfer learning for deep neural network based plant classification models”, Journal of Computers and Electronics in Agriculture, 2019
8. Jayme Garcia Arnal Barbedo, “Digital image processing techniques for detecting, quantifying and classifying plant diseases”, Barbedo SpringerPlus 2013, 2:660
9. Zhang Xiao, Yu Tan, Xingxing Liu, Shenghui Yang, “Classification method of plug seedlings based on transfer learning”, Appl. Sci. 2019, 9, 2725; doi:10.3390/app9132725
10. Catherine R. Alimboyong, alexander A. Hernandez, Ruji P. Medina, “Classification of plant seedling images using deep learning”, in: Proceedings of TENCON 2018- 2018 IEEE Region 10 Conference
11. Aharon Azulay, Yair Weiss, “Why do deep convolutional networks generalize so poorly to small image transformations?”,Journal of Machine Learning Research 20 (2019) 1-25

Tandem – repetitive minisatellite DNA portion of human can be used to produce someone’s specific Dna fingerprint. Studies shows the reliability of the analysis. Dna is the genetic fabric which is present in all the cells of the body. This molecule present a few characteristics, as VNTR, special present in different people. This can be a key in the improvement of few method, in this case DNA fingerprinting. This strategy has simple steps that we are going survey in this work. DNA fingerprinting procedure has gotten to be an imperative instrument for logical instrument. We’ll review few applications in areas like legal examination and parentage testing, additionally how this method has revolutionized and advanced in areas as Anthropological hereditary qualities, botony and zoology.[1] DNA process, one amongst the nice discoveries of the late twentieth century, has revolutionized rhetorical investigations. This review concisely recapitulates thirty years of progress in rhetorical DNA analysis that helps to convict criminals, pronounce the incorrectly suspect, and determine victims of crime, disasters, and war. [2]

INTRODUCTION

The technique well-tried applicable in several biological disciplines, particularly in diversity and conservation studies among species, and in clinical and social science studies. however verity political and social dimension of process became apparent so much on the far side educational circles once the primary applications in civil and criminal cases were revealed. forensic process is outlined because the comparison of the deoxyribonucleic acid in a person’s nucleate cells therewith known in biological matter found at the scene of against the law or with the deoxyribonucleic acid of another person for the aim of identification or exclusion. the applying of those techniques introduces new factual proof to criminal investigations and complaints. However, the primary case (March 1985) wasn’t strictly a forensic case however one amongst immigration.The primary application of deoxyribonucleic acid process saved a young boy from deportation and also the technique therefore captured the public’s sympathy. 1987 to the terribly initial application in a very forensic case. 2 young women had been raped and dead on completely different occasions in near English villages, one in 1983, and therefore the alternative in 1986. bodily fluid was obtained from every of the 2 crime scenes. The case was spectacular as a result of it amazingly excluded a suspected man, Richard Buckland, and matched another man, Colin Pitchfork, who tried to evade the DNA dragnet by persuading a friend to convey a sample on his behalf. Pitchfork confessed to committing the crimes once he was confronted with the proof that his DNA profile matched the trace DNA from the 2 crime scenes. The years once the invention of dna fingerprinting were characterised by a mood of cooperation and knowledge base analysis. None of the various young researchers who has been there’ll ever forget the biometric identification congresses that were held on 5 continents, in Bern (1990), in metropolis (1992), in Hyderabad (1994), in Melbourne (1996), and in Pt. Elizabeth (1999), and so stop working with the great feeling that the task was done.

The technological evolution of forensic deoxyribonucleic acid identification

In the classical deoxyribonucleic acid procedure technique radio-labeled DNA probes containing minisatellite or oligonucleotide sequences are hybridized to DNA that has been digestible with a restriction endonuclease, separated by agarose dielectrolysis and immobilized on a membrane by Southern blotting or – within the case of the oligonucleotide probes – immobilized directly within the dried gel. The radio-labeled probe hybridizes to a group of minisatellites or oligonucleotide stretches in genomic polymer contained in restriction fragments whose size differ due to variation within the numbers of repeat units. when washing away excess probe the exposure to X-ray film (autoradiography) permits these variable fragments to be pictured, and their profiles compared between people.[2]

SAMPLES FOR DNA FINGERPRINTING

Samples for typewriting of DNA will get from any diagnostic test tissue of a private, blood, saliva, duct fluid, semen, things of non-public use like razor, toothpick, bottles, cans or glass, used roll of tobacco butts, bite marks, bone teeth, hair, and garments or any variety of biological remains is used for identification. body waste also can be wont to extract the polymer from animal tissue cells that unleash from digestive tube throughout excreting however in recent studies it’s been shown that in healthy persons animal tissue cells can’t be discharged in excretion. just in case of blood white blood cells area unit beneath observation for attaining DNA as a result of these area unit nucleate cells. though red blood cells additionally contain nucleus however at maturation their nucleus disappears. Hair follicles bearing cells thus throughout investigation hairs act as proof. Our skin has higher layer of cells. This higher layer consists of keratinized cells that area unit dead in nature and shed off quickly from our body and act as proof at crime scene.

USES OF DNA FINGERPRINTING

It is wont to reason transmitted diseases, identification of victim (by comparison profiles of a victim with reference samples), usage for paternity test, process of a personal in a species, to visualize dissimilarities among people of various species, to search out out the organic process relationship among closely connected species, this system is employed in breeding programs, for following vulnerable species, used for sex determination, for conservation of variety.

AIMS

There are some objectives of this review that indicate the importance of this paper like to avoid negligence of handlers, with the assistance of DNA proof provide social control to a guilty person rather than innocent one, aim to pay full attention throughout a search work, another purpose is attempt to eliminate the incorrect perception of individuals concerning DNA finger-printing, taking a decent care throughout DNA proof analyzing, attempt to develop understanding among persons, unfold awareness to form a association between individuals and analyst and to boost the method techniques comprising in DNA analyzing method.

TECHNIQUES OF DNA PROFILING

Two styles of techniques are utilized in DNA typewriting, these are PCR (polymerase chain reaction) based mostly} and non-PCR based. Non-PCR based mostly techniques are ancient techniques, time taking, slow process, initiate sensible results and needed an outsized quantity of DNA sample for analysis. Whereas PCR based mostly techniques are fast in their nature and alittle quantity of DNA sample needed for analysis however there’s an opportunity of contamination throughout PCR amplification. Non-PCR based mostly technique is RFLP (restriction fragment length polymorphism). this method first of all employed by person within the laboratory throughout analyzing DNA. On the opposite hand, STRs (short cycle repeats), microsatellites, and mini-satellite are PCR based mostly techniques. of these procedures establish VNTR’s (variable range of cycle repeats) regions within the genome. There are such a lot of different techniques for analyzing DNA process however these don’t offer correct results. The Y-chromosome technique is a complicated technique however it’s utilized in extremely economical laboratories. If the DNA samples are shopworn like hair, tooth, and bone then it lacks DNA within the nucleus or improper DNA is present that is tough to judge. therefore RFLP and STR technique weren’t applicable as a result of they need organelle DNA. during this case, mitochondrial DNA is used for genetic interpretation. applied science, DNA microarray analysis and multiplex PCR technology are advance techniques in DNA identification however these techniques face some challenges in identification of DNA. dielectrolysis gel, southern blotting and interbreeding additionally wont to transcribes the results. PCR technology additionally produce to errors like stature formation, slippage of primer and false peaks are shown on the graph differing types of DNA techniques area unit utilized in assortment of DNA sample name as static Detection equipment (ESDA), dry swabbing and substrate cutting. ESDA and dry scrub are non-destructive whereas substrate cutting may be a damaging technique .Another technique that is known as “Real Time PCR”, it’s wont to confirm the gender from a degrade DNA sample SRY PCR is additionally a sex confirm technique. [3]

CONCLUSION

DNA process is obtaining larger and a lot of universally accepted with time. Since nobody will alter their deoxyribonucleic acid sequence when going it at the crime scene and since it’s exhausting to stop effort one’s deoxyribonucleic acid at the crime scene, deoxyribonucleic acid analysis is arguably the best forensic tool employed in forensics. Of the 3 main forms of deoxyribonucleic acid fingerprints, RFLP, VNTR and STR, the foremost normally used is that the STR.

Restriction fragment length polymorphism and VNTR need a great deal of deoxyribonucleic acid, that is sometimes terribly tough to search out at the forensic scene and infrequently the deoxyribonucleic acid fragments being analyzed are too long to amplify via PCR. STR, on the opposite hand, uses short sections of deoxyribonucleic acid, that are ideal for running a PCR. there’s associate exponential increase of the degree of deoxyribonucleic acid, so creating it easier to run tests on little samples of deoxyribonucleic acid. in addition, STR analysis doesn’t need the hybridizing to a deoxyribonucleic acid probe, which might are long. Forensics has taken an outsized step exploitation deoxyribonucleic acid to resolve crimes, that were insolvable within the past. As deoxyribonucleic acid is personal, it’s extraordinarily rare that the deoxyribonucleic acid can match over one person on this planet. This has allowed deoxyribonucleic acid proof to be accepted within the court space.[4]

Languages are vehicles for the transition of cultural knowledge and act as a lens by which individuals view the world. Linguistic diversity is threatened by language contact due to invasion and colonisation and more currently the forces of globalisation and language homogenisation (Zuckerman & Walsh, 2015). Seven thousand languages are spoken in the world today however nearly half (3 billion) of the world’s current population speak one of only twenty languages as their first language (Austin & Sallabank, 2015). More than forty percent of these 7000 languages are in an endangered state (‘Endangered Languages Project’, 2017). UNESCO defines a language as endangered when it is no longer used by speakers for communicative purposes and intergenerational transmission has ceased (UNESCO Ad Hoc Expert Group on Endangered Languages, 2003).

Many language revival and language revitalisation projects have been established by linguists, academics and community leaders in the hope of slowing down language loss processes. Language loss occurs within communities that have lost their last generation of native speakers. Language revival programs aim to renew these languages that have experienced language loss. Whereas, language revitalisation programs attempt to reinvigorate languages that are still being partially acquired through intergenerational teaching but are in an endangered state. The current fragile state for minority languages creates an environment where preparation for the eventual loss of language experts is necessary for linguistic survival. Language communities preparing for this transition can derive strategies from previous successful language revival programs and analyse the mistakes from unsuccessful language preservation attempts. This essay will explore the successful language revival program utilised to inject new life into the Hebrew language alongside the language revitalisation of Gumbaynggir. The many attempts to revitalise the Irish language illustrate ineffective strategies that when analysed give insight into whether certain programs have the ability to improve or should be avoided entirely.

In order to revive or revitalise a language, conservational linguists must first explore the factors causing the language to be endangered. Languages can become endangered due to communities being in physical danger and losing members during natural disasters, war and genocide. This is exemplified by language death due to genocidal violence towards the Palawa people during the ‘Black War’ (Austin & Sallabank, 2015). Over time, marginalization of a language community due to cultural or political pressure can discourage use of a language and cause forced linguistic assimilation. Direct repression by legislation restricting use of minority languages or indirect pressure due to economic advantage of a majority language may trigger language abandonment. These factors can be a catalyst for language shift within a minority language community as native speakers no longer use the language and intergenerational transmission processes discontinue. Once factors for language endangerment have been explored processes to actively preserve and revive the language can begin.

Firstly, conservational linguists must work alongside native speakers to document the language within dictionary corpus, extensive grammars and archives of traditional stories and songs. Modern language documentation processes can positively utilise technology as shown within the Gamilaraay community. Monetary issues due to lack of funding, and social stigma associated with learning the language can be overcome with use of digital platforms including online tools and social media. Interactive games, visual aids such as animations and audio recordings are used to learn and document the language (Smith, Giacon & McLean, 2017). Video and audio recordings of native speakers preserves vital linguistic information about the accent, tone and syntax used within the language that cannot be documented by text. Facebook groups allow community members to connect and share their language learning journey creating a sense of community and inspiration for further programs. These digital platforms that have previously been viewed as a danger to minority languages have potential for success in language revitalisation programs in the future (Jany, 2017).

The British conquest and colonisation of Ireland began a forced language shift towards English and a decline in the number of native Irish speakers. English was the majority language spoken by the middle of the 19th century, with Irish continuing to decline until recently. In the 2016 census 1.7 million people answered yes to being able to speak Irish. However, the “self-reporting” nature of the census reduces accuracy of these results as individuals may report to speak Irish whether they are fluent or if they only know a few phrases. Further questions revealed that only 74,000 individuals spoke Irish daily (Irish Central Statistics Office, 2016). Within the Gaeltacht (Irish-speaking region) 66.3% of the population could speak Irish, with only 21.4% speaking Irish daily. The number of Irish speakers in the Gaeltacht has declined since 2011 by 2.2%. Governmental structures, individuals and organised groups have been taking measures in an attempt to revive the Irish language since the middle of last century. The Irish language movement is prominent in the literature and is often used as an exemplar of language revival failure.

Official structures such as the Irish government and the European Union (EU) have implemented many strategies in an attempt to revive the Irish language. In 2005 Irish was declared as an official language of the European Union and is currently the least spoken EU official language (‘Working languages of the European Parliament’, 2019) The Irish government took a two-step approach, firstly incorporating Irish into the education system, to encourage use in the English speaking region. Secondly, language maintenance measures were established to protect the Gaeltacht community. Governmental language revival policy relied heavily on the National School system. Fr. Timothy Corcoran of the Irish National Teacher’s association believed that Irish could be restored by the school system, “even without aid from the home” (Titley,1983). Initially, Irish was used at the infant level in some classes, however over time Irish became the medium of instruction during all classes. The amount of Irish used was at the discretion of each school, with 704 schools using exclusively Irish in 1939 (MacNamara, 1996). The use of Irish within the school setting was found to be detrimental to children’s learning of English and showed no improvement in Irish proficiency. Infants were no longer taught in Irish in 1970, and by 2016 ordinary schools had significantly reduced their use of Irish in the classroom setting. (Edwards, 2010) Incorporation of Irish into the education system was ineffective in reversing language shift. Use of Irish in a classroom setting does not transfer to Irish use in the home or wider community. Further to this, children forced to learn the language in school, or through external classes may hold feelings of antipathy towards the language. Such feelings reduce the likelihood of such individuals encouraging their own children to learn the language. Social interactions, workplace communication and wider media are all English- based leaving no communicative function for Irish. Without a communicative function the patriotism surrounding Irish is not enough of a motivating factor to save the language.

Widespread emigration from the Irish speaking region due to the isolated geographic location, and economic weakness of not only the region but the language itself. The Gaeltacht has a low employment rate due to lack of urbanisation. As English is the prestige language speakers of the Irish language are disadvantaged with many high-ranking positions requiring English shifting the language of the workplace (Carnie, 1995). Until 1973 proficiency in Irish was required for employment in the Police force, army, legal profession or civil service (Edwards, 2010). Whilst this could be beneficial for Irish use, in reality majority of personnel hardly used the language. Another issue threatening the longevity of the Irish language is the ageing population of Irish speakers within the Gaeltacht. In order to reduce emigration from the Gaeltacht the government gave a small grant to Irish speakers called the deonatas. Whilst this was motivating, the Údarás na Gaeltachta department was established to overlook sociocultural and monetary aspects of the Gaeltacht. (Údarás na Gaeltachta, 2019). Economic investment into the creation of jobs in order to increase the employment rate and retain Irish speakers was successful. There was a subsequent population rise, however not necessarily of Irish speakers. It is vital for the survival of the language that the Irish speaking population of the Gaeltacht is stabilised. Without that population the future of Irish will be unsteady and continue declining.

Hebrew had become a heritage language not spoken since the Roman empire and only used in situations of written prayer. Millions of people from different linguistic backgrounds, mostly holocaust survivors, participated in a mass migration to Israel. The establishment of this Jewish Nation required a national language and Hebrew, the language of prayer was deemed the appropriate choice. Modern Hebrew was invented by Eliezer Ben- Yehuda, speaking to his son only in Hebrew creating words for modern items and ideas. The language sprung to life with the first native speakers in 1,800 years and continues to be strong to this day with 9 million Hebrew speakers (Bensadoun, 2010). Many linguists claim this impressive revival to be the most successful recovery of a “dead” language to date. Therefore, revival linguists can investigate and compare strategies implemented during the revival of Hebrew to current and future revival programs. Firstly, there is extensive and exhaustive language documentation of Hebrew in religious literature including the Hebrew Bible and Misnah. As Hebrew is already present within the community, positive language attitudes have already been established. A level of prestige and familiarity is associated with the Hebrew language due to literature exposure in the synagogue. Such language attitudes help ease the community into the language change alongside other contextual factors associated with the community. Indigenous Australian language revitalisation programs should adopt certain positive attitudes towards language revival attempts used in the Hebrew revival. An openminded approach towards the emerging hybrid language should be adopted. When a communicative need is no longer fulfilled by the language it must be flexible and adaptative in order to survive. Influences from surrounding languages including English on the languages grammatical form or the introduction of modern words and concepts should be accepted.

Further to this, acceptance of the current state the language is in by community members is needed before the process of language revitalisation begins. For example, some Gumbaynggir community members objected to the claim that “most Indigenous languages in ‘settled’ Australia are extinct” saying that “their language has never died” (Walsh, 2001). Community leaders can use this tenacity to inspire such individuals to assist with the revitalisation program. These pre-revival approaches align with those used by the Hebrew revival that emphasise “observation, documentation and characterisation of the community”. Currently many revitalisation movements are under indigenous community control run with the help of conservational linguists. This context applicable both to Indigenous and the Hebrew community creates an environment for self-respect and empowerment however sustained commitment from external funding sources must be obtained to ensure longevity. The geographically isolated nature of rural indigenous communities prevents cohesion and sharing of knowledge learnt from successful and unsuccessful programs. Annual conferences of all regional programs, an Australian language revival handbook, and use of social media may be helpful resources for programs to implement in the future.

Within the 2015 film, The Martian, the main protagonist is able to grow food sources on Mars when he is left after complications in a NASA mission. The capability of sustainable plant growth, to the quality seen on Earth, is questionable. This is where the question, ‘Are the parameters of Mars suitable for sustainable plant growth and survival?’ was asked.

To come to a conclusive answer for this question, it would need to be testable. The testability of this question can be decided when it is possible to be proven or disproven, if a relationship can be demonstrated within two things, that is the plant growth and the environmental factors, and it’s independent variable and dependent variable are all taken into consideration. The confirmation of this information and whether or not the answer can be considered fact is based on the collection of data, and the repetition of an experiment to minimise outliers and get a more accurate answer. Each variable and process involved within the testing of this question need to be capable of being reproduced, strictly following the steps taken.

This question is capable of being reproduced, this is due to the necessary variables for plant growth on Earth can be tested and comparing these factors to the only information on Mars’ variables so it is comparative to the same set of data.

What is actually needed for a plant to thrive? A multitude of factors go into developing healthy plant growth. To begin with, water, fresh clean air and healthy soil that is nutrient dense is essential for growing healthy plants to the best of their potential. Air that lacks oxygen, and the dust levels that Mars’ has is harmful to plants as it limits their ability to absorb carbon dioxide. This is impactful as it inhibits the process of photosynthesis, where plants make their own food to grow. Healthy soil is also extremely vital to plants as the essential nutrients found in soil help to anchor plant roots and support the plants through their growth. Temperature and sunlight also impact the plant, as exposure to sunlight aids in the process of photosynthesis, and temperatures are important as too low can freeze the plant and too high may burn it. All of the aforementioned contribute to ideal plant growth in a particular way.

There are significant quantities of water on the ‘Red Planet’ although it is dry by terrestrial standards, all this water is ice just below Mars’ surface. Many people believe that due to there being a high content of water on Mars, especially in it’s Martian soil that planting a seed will turn into a plant, but there is more needed than water for plants to grow.

According to the U.S. Department of Agriculture’s Natural Resources Conservation Service, on Earth it takes 500 years to develop only one inch, or approximately two and a half centimetres of topsoil. This topsoil is filled with living and dead organisms – including plant matter, bacteria, rocks and nutrients such as nitrogen, all of these components aiding in ideal plant growth. As far as released research shows, Mars does not host these organisms to develop nutrient dense topsoil the same way that Earth does. In saying this, Martian soil has been found to contain specific nutrients that are beneficial for plants.

According to space.com Mars’ atmosphere is 100 times thinner than that on Earth, without that atmospheric thermal blanket retention of warmth is impossible. On average, Mars’ temperature is minus 60 degrees celsius, optimal temperatures for plant growth are around 25 degrees celsius, meaning that seeds will freeze before seedlings or plants even get the chance to grow. Even taking into account Mars’ temperature on a hot Summer’s day (closer to the equator) temperatures only rise to about 20 degrees celsius, while at night they can plummet to minus 100 degrees.

Sunlight, is also a factor that must be taken into account for plant growth, due to the process of photosynthesis. At the Earth’s surface, with the Sun directly overhead at local noon (clear dry atmosphere), the solar irradiance is about 1000 W/m2 (1000

watts per square metre). This value is highly variable depending upon such things as the amount of dust and water vapor in the atmosphere. At local noon on Mars, with Sun directly overhead, the solar irradiance is 590W/m2 (590 watts per square metre). This wattage means that noon in Mars the sunlight is the same level it would be at 2.2 degrees celsius on Earth. This means that sunlight levels on Mars are not sustainable for a natural photosynthesis process to occur within any plants that could hypothetically survive all of Mars’ other harsh conditions.

According to the main qualitative data we have surrounding both plant growth and Mars’, all of which previously aforementioned we can see that when they are compared plant growth on Mars is quite simply unsustainable.

Granting all this, Martian soil is nutrient dense and various plants have been grown using the soil. Martian soil contains macronutrients such as oxygen, nitrogen, potassium and phosphorus. It also contains micronutrients such as iron, manganese and boron. Although there may not be the right amount of nutrients depending on where astronauts land on Mars, so fertilisers may need to be added to the soil. If we were to grow plants on large sections of Mars, the amount of fertiliser required would be excessive and the cost of transferring it from Earth would be too. Real Martian soil is also perchlorate. Perchlorates abound in the uppermost layer of Martian regolith, potentially lowering the freezing temperature of water – enough to explain some fleeting signs of liquid water activity on Mars.

Perchlorate is toxic to humans, causing thyroid problems and even death. If we were to move to Mars, breathing it in from the dust that could infiltrate any develop habitats and growing food from this soil would be extremely dangerous. This is where the main protagonist of the 2015 film ‘The Martian’ would have died.

If we genuinely wanted to grow plants in Martian soil, whether that is on Earth or on Mars the removal of perchlorates would be essential to allow the edibility of the plants grown to be maintained. In small portions, this can simply be done by thoroughly rinsing the soil in water, as these perchlorates are water soluble. Although, if we were to realistically look at growing plants on Mars, such a large amount of soil would require something on a larger scale and definitely more intriguing – the introduction of perchlorate-eating bacteria of which produces oxygen as a metabolic byproduct. Yet, even the bacteria would have its own queries, such as if it was able to survive the harsh conditions of Mars, how much of this bacteria we would have to introduce and whether it is capable of removing this perchlorate from each layer of Martian soil. We could also question if the oxygen released by the bacteria would sustain within the Martian atmosphere to develop air that would be similar to that on Earth, especially due to Mars having 95% carbon dioxide in its atmosphere.

Although there is an abundance of water on Mars, it is in the form of ice. When developing the theoretical hydroponic greenhouse to maintain heat and grow crops after altering Martian soil, the ice could be melted and used to water these plants. Except, since the Martian soil is so dry, the watering of plants would be increased severely to maintain enough water to suffice for the plants to avoid drying out. For sustained growth, solutes must continuously be imported into growing cells so that the osmotic potential does not increase in the face of osmotic dilution by inflowing water. Solute import thus assures maintenance of the water potential gradient that will sustain inward water movement. Since Mars has a very thin and cold atmosphere, plants grown on the planet would need to be in a greenhouse. This isn’t inherently a bad thing — Guinan said in an interview with the American Geophysical Union that the greenhouse plants could be part of the atmospheric recycling process for the idea of a Martian colony.

In essence, when we compare qualitative and quantitative data supplied by articles and credible scientific sources – as well as logically looking at the current state of Mars we can come to the conclusion that healthy, natural plant growth on Mars is simply not possible. We also must take into account the credibility of our sources, as for a lot of us primary research of Mars is not possible. A portion of data that has been produced throughout this literary reflection comes from NASA. NASA is a credible source of information, NASA knows that if it is found to release information that is false whether they partner with a corporation or otherwise the media attention will strip their credibility and remove funding after their reputation would be tarnished as such. NASA is purely scientific and wants to aid the scientific world such as the ISS, moon landings, and other missions, as its purpose is to gather data for scientists and encourage the public to be active and conscious about science. Scientists are currently working on ways to alter Mars to make it more suitable, as well as altering the plant matter itself to suit Mars and of course one day there is always the possibility of making this happen although it would be years, possibly decades away and at what cost?

BIBLIOGRAPHY

1. Carlisle, C. M. (2018, January 17). Some Plants Grow Well in Martian Soil. Retrieved July 12, 2019, from https://www.skyandtelescope.com/astronomy-news/some-plants-grow-well-in-martian-soil/
2. Dunbar, B. (n.d.). Designer Plants on Mars. Retrieved June 29, 2019, from https://www.nasa.gov/centers/goddard/news/topstory/2005/mars_plants.html
3. Humphries, P. (2019, July 17). Micro-managing the Murray-Darling Basin: What’s in it for fish? Retrieved June 28, 2019, from http://theconversation.com/how-to-grow-crops-on-mars-if-we-are-to-live-on-the-red-planet-9994
4. Jordan, G. (2015, October 05). Can Plants Grow with Mars Soil? Retrieved July 25, 2019, from https://www.nasa.gov/feature/can-plants-grow-with-mars-soil
5. Plant Growth. (n.d.). Retrieved July 6, 2019, from https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/plant-growth
6. Tests Indicate Which Edible Plants Could Thrive on Mars. (2018, January 12). Retrieved July 22, 2019, from https://eos.org/articles/tests-indicate-which-edible-plants-could-thrive-on-mars
7. The experimentation of growing plants on Mars. (n.d.). Retrieved July 1, 2019, from https://www.jhunewsletter.com/article/2018/10/the-experimentation-of-growing-plants-on-mars

The higher the temperature the faster the particles move. This is because of the kinetic energy, the more kinetic energy there is, the greater the chance is for a collision that results in a reaction. More energetic particles have a higher chance of “COLLIDING SUCCESSFULLY because they will collide more often thus increasing the number of successful collisions PER SECOND, thus increasing the rate of reaction”.

Concentration

“As the concentration of a reactant/s increases, the rate of reaction increases”. This is due to the big amount of reactant particles, this also means that they are very close together, so the “frequency of collisions increases”. Thus, leading to an increased rate of reaction. Socratic. (2014, August 7)

The bigger the surface area of a reactant, the more the reaction rate occurs”. This is because of more particles being exposed, increasing the surface area available for collisions to take place. This also does ‘not’ affect the energy of the particle.

Catalysts

A catalyst is a special chemical that speeds up the rate of reaction but is not consumed by the reaction. This allows it to only use a small amount of energy to activate and comes out of the process unchanged. This means that more particles collide with enough energy to cause a collision; effectively increasing the collision rate. All catalysts will speed up the decomposition on H2O2. The most effective, based on the background research, will be the potassium iodine.

A chemical bond forms when the valence electron of two or more atoms interact. Thus, electrons are either shared or transferred between the atoms, making them more stable. The making of chemical bonds releases energy because of the bond making the atoms more stable (2020). However, breaking chemical bonds require energy because separating them makes them less stable. Chemical reaction occurs due to bonds between atoms either forming or breaking. This meaning that two or more molecules have interacted and caused the molecules to change. However, in some cases molecules can repel each other, this is caused by two molecules of the same charge interacting (2020). Energy plays a significant role in chemical reactions as chemical energy is stored in the bonds of a chemical compound, like atoms and molecules. Energy can be added or released from a molecule by the rearrangement of electrons. The releasing of a chemical energy can also cause the substance to turn into a completely new substance. If a chemical reaction released energy, either light would be seen, or the beaker would start to heat up (2020). The amount of heat released can range from slight warming of the beaker to an explosion.

Energy comes in many forms and is the ability to do work or cause change. The usage of energy is the process of transferring energy from one thing/place to another. There are two different forms of energy -kinetic energy and potential energy. Kinetic energy is associated with motion. As such all moving objects have kinetic energy however, the amount in each object differs (‘Kinetic particle theory – Kinetic particle theory and state changes – GCSE Physics (Single Science) Revision – Other – BBC Bitesize’, 2020). The amount of kinetic energy in an object is determined by the mass and its speed. The heavier and faster the object the more kinetic energy it has. On the other hand, potential energy is ‘stored energy’. “It is an energy that remains available until we choose to use it. There are a lot of different ways in which energy can be stored which makes it difficult to recognise potential energy”. An object has potential energy because of its position relative to another object. Another way for energy to be transferred is through heat or work. Heat is the term given to energy that is transferred from a hot object to a cooler object, however, it will stop when it has reached an equilibrium. Work, however, is energy transferred as a result of a force applied over a distance.

Some laws apply to energy, the law of thermodynamics being one. The first law is that energy cannot be created nor destroyed, but it can be transferred or transformed. The second law states that energy transfers and transformations increase the entropy of the universe. Entropy is the functions that describe the level of disorder in a thermodynamic system (‘Enthalpy and Entropy | A-Level Chemistry Revision Notes’, 2020).

There are two types of energy changes in a chemical reaction. One of them is called exothermic reaction and the other is Endothermic reaction. Endothermic is heat energy absorbed from the surroundings. The energy level in the product is higher than the energy level in the reactants. Endothermic is the releasing of heat energy into the surroundings (2020). Consequently, the energy level of the product is lower than the energy level of the reactants. The result of an exothermic reaction is having products with more stable bonds (lower energy), whilst endothermic reaction results in the product with a light energy (less stable) (‘Home’, 2020).

Discussion

The experiment, elephant’s toothpaste relates to the concentration of the reactants because the hydrogen peroxide is being diluted every time. The different concentrations in the solution has led to great findings and ideas for future investigations. A major finding was that the 3% solution, which contained 10ml of water, lasted for 3min and 35 seconds. The experiment consisted of decomposing Hydrogen Peroxide, Potassium Iodine and the water acted as a catalyst.

This experiment allowed the hypothesis to support it. The 3% solution was a diluted form, meaning that there were less hydrogen peroxide particles. Due to the little number of particles, the reaction was much slower. This had resulted in the long reaction rate. This was due to there being an insufficient number of hydrogen molecules (unstable) to decompose (‘Home’, 2020). Due to the small opening in the test tube oxygen is slowly being released into the surroundings, which can also cause the slow decomposition. Another reason can be the temperature, as the temperature in the testing room was neutral, of this was not the case then the temperature would have acted as a trigger for thermal decomposition (‘Chemical reactions | Chemistry of life | Biology (article) | Khan Academy’, 2020).

[bookmark: _Toc35179340][bookmark: _Toc35192628][bookmark: _Toc35258550][bookmark: _Toc35466003]Through the process a problem has been encountered, measurement of the solutions. When conducting the 12%-3% the amount of mixture within the tubes were different from one another. This has caused the results to be skewed, which had called for a redo of the entire produce. The second take of trial 1 and 2 has allowed the result to be more accurate and credible as a result of all the solutions being of the same amount. An improvement that would have been useful in this experiment would have been to record the temperature. This could have explained the reason for the chemicals to react in a certain manner. Further investigations can include comparing the reaction of the elephant’s toothpaste to another experiment similar to it. Or adding the detergent in and seeing if this can affect the reaction rate.

It has been stated that the use of inorganic catalyst such as manganese oxide or lead can cause the decomposition of the hydrogen peroxide to happen faster. However, would the concentration of the hydrogen peroxide affect the reaction rate? Other ideas could include using different temperatures of water as temperature affects the reaction rate. The experiment could also be tested in the sun, as exposure to the light would have decomposed it quicker (Socratic. (2014, August 7). However, the question is if whether having both the warm water and sun acting as catalyst with only 3% hydrogen peroxide can cause an entirely different reaction. This may cause a different reaction due energy that is either absorbed or released.

Energy is related to the experiment ‘Elephants toothpaste’ since the decomposition of hydrogen peroxide releases energy in the form of heat. As heat is released into the surrounding areas the breaker/tube the experiment was conducted in would start to heat up (reaction?,2020). Thus, causing an exothermic reaction (Foundation, 2020). The conduction of the experiment, elephant’s toothpaste has shown how the decomposition of hydrogen peroxide work. It has also allowed numerous questions to be asked, which could be possible ideas for future endeavours.

Conclusion

Overall, the elephant’s toothpaste experiment has allowed the hypothesis supports it. The results clearly show which concentration has the longest reaction rate. Many things that makes this possible is the less amount of hydrogen peroxide particles and the high amount of oxygen left. these variables have caused bubbly and heat as energy is transferred from the solution to the glass. Though there was an error which skewed the results, correct measures were taken to make the results once again credible. The task of designing and carrying out an experiment that related to the concentration of the reactants has been completed successfully.

Bibliography

1. reaction? W. (2020). What are the two types of energy changes that can occur in a chemical reaction? | Socratic. Retrieved 18 March 2020, from https://socratic.org/questions/what-are-the-two-types-of-energy-changes-that-can-occur-in-a-chemical-reaction
2. Chemical Energy – Knowledge Bank – Solar Schools. (2020). Retrieved 18 March 2020, from https://www.solarschools.net/knowledge-bank/energy/types/chemical
3. Foundation, C. (2020). Exothermic and Endothermic Processes. Retrieved 18 March 2020, from https://www.ck12.org/chemistry/exothermic-and-endothermic-processes/lesson/Conservation-of-Energy-in-Chemical-Reactions-MS-PS/
4. Chemical reactions | Chemistry of life | Biology (article) | Khan Academy. (2020). Retrieved 18 March 2020, from https://www.khanacademy.org/science/biology/chemistry–of-life/chemical-bonds-and-reactions/a/chemical-reactions-article
5. Movement of particles. (2020). Retrieved 18 March 2020, from https://www.education.vic.gov.au/school/teachers/teachingresources/discipline/science/continuum/Pages/particles.aspx
6. (2020). Retrieved 18 March 2020, from https://orise.orau.gov/stem/documents/k-12/lesson-plans/energy-from-chemical-reactions.pdf
7. (2020). [Image]. Retrieved from https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.slideserve.com%2Famy%2Felephant-toothpaste-physical-or-chemical&psig=AOvVaw14heJTRk6jolU1Z8wJ1XzA&ust=1584608851616000&source=images&cd=vfe&ved=0CAIQjRxqFwoTCMDNuJTWo-gCFQAAAAAdAAAAABAd
8. catalyst | Examples, Definition, & Facts. (2020). Retrieved 18 March 2020, from https://www.britannica.com/science/catalyst
9. (2020). Retrieved 18 March 2020, from https://orise.orau.gov/stem/documents/k-12/lesson-plans/energy-from-chemical-reactions.pdf
10. (2020). Retrieved 18 March 2020, from https://www.abc.net.au/science/surfingscientist/pdf/teachdemo26.pdf
11. (2020). Retrieved 18 March 2020, from https://www.bcsoh.org/cms/lib/OH01001261/Centricity/Domain/763/Chemical%20Energy%20tutorial%20KEY.pdf
12. Enthalpy and Entropy | A-Level Chemistry Revision Notes. (2020). Retrieved 18 March 2020, from https://alevelchemistry.co.uk/notes/enthalpy-and-entropy/
13. Entropy. (2020). Retrieved 18 March 2020, from https://simple.wikipedia.org/wiki/Entropy
14. (2020). Retrieved 18 March 2020, from https://study.com/academy/lesson/entropy-in-chemistry-definition-lesson.html
15. Kinetic particle theory – Kinetic particle theory and state changes – GCSE Physics (Single Science) Revision – Other – BBC Bitesize. (2020). Retrieved 18 March 2020, from https://www.bbc.co.uk/bitesize/guides/zq7t4j6/revision/1
16. What does hydrogen peroxide spontaneously decompose into?: Socratic. (2014, August 7). Retrieved from https://socratic.org/questions/what-does-hydrogen-peroxide-spontaneously-decompose-into
17. Home. (n.d.). Retrieved March 2, 2020, from http://scactivities.cikeys.com/elephant-toothpaste/background/

Abstract

The exploitation of shark species to satisfy the high market demand is now becoming a worldwide concern since the numbers for many species of these apex predators are now declining at an alarming rate. Shark DNA extracted from shark samples collected from the Australian fisheries management, amplified and used for identification. Although there are different methods of identifying a species, they proved to be insignificant at times of need. A dire need for species identification method that is both accurate and reliable was needed. It was established that the mitochondrial gene cytochrome oxidase I (COI) could provide a viable way for bio identification for animals. This study was performed with the aim to better understand the rate of illicit shark catches and more importantly stop illegal fishing of endangered shark species and secondly to check the reliability of the DNA barcode method using the COI gene. The result obtained showed that the samples were properly identified to the species level, with profound accuracy.

Introduction

The increase in the popularity of shark fin soup worldwide also increases the threats of illegal fishing of some endangered species of these cartilaginous fish. Two other factors can also be an indirect threat to the shark species which is, mislabelling of shark meat and unnecessary shark fishing. The overfishing of sharks is specifically tricky due to the fact that these predators present a key function in the marine ecosystem, and, therefore, their population dynamics can also have an effect on all local marine diversity (Bonfil, 1994).

And previous studies reveal that the captured shark is usually found without their fins and their head removed and the body is sold in the market without proper labelling or just as ‘Sharks’. This inhibits the tracing of the structure of the species and recognition of the sharks which are protected by the laws of fisheries. The great white, grey nurse & Bull Shark has been marked as a near threatened or endangered shark species by IUCN (International Union for Conservation of Nature) (Diaz-Jaimes et al., 2014). In numerous situations, sharks have also been the focus of risk reduction programs owing to attacks on humans (Dudley and Simpfendorfer, 2006). Although sharks have been used commercially for its skin, liver oil and flesh, at present its fins are the major product that increases the demand for this and many other species. To stop this illicit shark fishing for the fin and other shark by-product and save the near-threatened or endangered species the identification of species presents a vital opening in stopping these businesses (Bornatowski, Braga and Vitule, 2014).

It was necessary for a quick yet effective testing process to specifically identify shark species. And so, the development of DNA barcoding revolutionized the way of species identification. The cytochrome Oxidase I (COI) gene, of the mitochondrial DNA (mtDNA), is a consistent gene section that allowed researchers a quick and exact identification of species (Hebert, Cywinska, Ball and deWaard, 2003) (Holmes, Steinke and Ward, 2009). Earlier methods of testing included the use of polymerase chain reaction (PCR) assays and species-specific primer (Magnussen et al., 2007) (Clarke et al., 2006) (Shivji, Chapman, Pikitch and Raymond, 2005).

Materials & Methods

The unidentified shark tissue sample for this study was collected from the apprehended illegal catches, received from the Australian fisheries management, these samples were either seized from poachers’ boat or found as finless and headless shark carcasses by the sea-shore or at sea by the management. An approximate of 16 tissue samples were collected (Table-1). The tissues were properly labelled and stored in 90% ethanol at a temperature of -20oC. The DNA extraction for each sample was carried out using the salting out method and the extracted DNA was stored at a cool temperature until further use (Armani et al., 2015).

DNA extraction

For DNA extraction 2 mm3 of the tissue from every sample was used. The tissues were lysed in a 1.5 ml microcentrifuge tube using 20µl Proteinase K (Sigma Aldrich Australia) and 580µl TNES buffer and left at 55oC for 3 hours. The proteins were precipitated with 170 µl 5M NaCl and vortexed for 15 seconds before centrifuging for 10 minutes at 14000 rpm. The supernatant liquid containing the DNA was transferred to a new microcentrifuge tube containing 770 µl of ice cold 100% ethanol. The precipitated DNA were extracted, washed, air dried and diluted with 30μl sterile water & kept at a cold temperature prior to PCR (Aljanabi, 1997) (Sunnucks and Hales, 1996). The final product contained 30μl of resuspended DNA stocks from which, DNA fragments were evaluated by gel electrophoresis on 2% agarose gel and 10 µl of the DNA was stained with GelRed (Sigma Aldrich Australia) and DNA extraction success was examined (Figure-1). The gel was run at 110V for 30 minutes.

PCR amplification and sequencing

For amplifying the COI gene, instructions by the manufacturer were followed for PCR, where 5μL of the DNA stock was added to 95μL of sterile water in a 1.5 ml microcentrifuge tubes to get a 20-fold dilution. In a 0.2 ml PCR tube, 2μL of the diluted sample was added to the master mix. The content of the master mix includes 10µl qPCR mix Kappa SYBR fast (Kapa Biosystems, United States) containing dNTPs, MgCl2, and Taq polymerase and forward primer and reverse primer of 0.5µl each with 7µl of sterile water for PCR. The COI fragment would be 650 bp long. The cycling conditions were set to 35 cycles having 950C for 1 min, 940c for 30 sec, 520c for 30 sec, 720c for 1 min and a final extension of 720C for 10 min (Ward et al., 2005). The amplified COI genes were identified using Sanger Sequencing and the instrument used was Macrogen Inc. (South Korea). The chain-terminating inhibitors dideoxynucleotide (ddNTP’s; defective A, T, C, G) were used for the sequencing (Sanger, Nicklen and Coulson, 1977). Prior to PCR analysis 5μL of the PCR products were run into 2% agarose gel and compared with a 100bp DNA ladder to assess the presence of expected amplicons which were later amplified (Armani et al., 2015). The gels are viewed using the BIO-RAD Gel Doc EZ imager.

Data analysis

The sequence obtained from the PCR were aligned using ClustalW integrated into MEGA X (Kumar et al., 2018) (Tamura et al., 2013). The sequences were trimmed manually in the MEGA X after visual inspection and they were used to run a BLAST analysis and sequences that showed 100% perfect similarity on GenBank were selected for possible species identification (Barbuto et al., 2010). The sequences were deposited into the BOLD system (Barcode of Life Data System) for identification (RATNASINGHAM and HEBERT, 2007). A phylogenetic tree (Figure-4) was constructed using the MEGA X with neighbour-joining method using the bootstrap option to calculate node statistics (Zhang and Sun, 2008) (The neighbor-joining method: a new method for reconstructing phylogenetic trees., 1987).

Discussion and Conclusion

The use of the COI gene produces more reliable results and allowed the identification of all the shark samples at the specific species level. However, the success of this study is dependent on factor such as DNA quality, as a result, some samples that had poor DNA quality was reprocessed before PCR amplification and sequencing was performed.

There are other molecular methods for shark species identification. One of them includes the design of species-specific primers to identify species of sharks. Advantage of this method includes identifying multiple species even in a processed or degraded sample, but the shortcoming of this process is the insufficient collection of species-specific primers and the probability that the primers may also not be one hundred percent specific to the sample DNA (Abercrombie, Clarke and Shivji, 2005) (Hoelzel, 2001).

The results from the study suggest alarming outcomes about how demanding shark products are which is becoming the driving force for these illegal fishing businesses. These situations were already reported in other studies (Ward et al., 2005), and reported as shark carcasses being misidentified and incorrectly labelled. The results from table-1 indicate how the popularity of shark product globally causing fishers to extensively and illegally hunt sharks without proper controls and possibly bringing an upset to the marine ecosystem by

Identification using the distinct DNA barcodes can be a practical way to monitor mislabelled shark carcasses and prevent the extinction of these apex predators who play a major role in the marine ecosystem. The worldwide exhaustion of the shark population that has been documented universally (Ward et al., 2005) indicates that these species require critical attention and conservation. Additionally, proper monitoring of their genetic stock is particularly important to secure the chances of the full recovery and maintenance of the genetic diversity.

It is conclusive that DNA barcoding can be applied to identify shark species and is a reliable and rapid method that fisheries management authorities can utilize to collect data from legal and illegal shark fisheries and manage species conservation.

References

1. Bonfil, R., 1994. Overview Of World Elasmobranch Fisheries. Rome: Food and Agriculture Organization of the United Nations.
2. Diaz-Jaimes, P., Uribe-Alcocer, M., Hinojosa-Alvarez, S., Sandoval-Laurrabaquio, N., Adams, D. and García De León, F., 2014. The complete mitochondrial DNA of the bull shark (Carcharhinus leucas). Mitochondrial DNA, 27(1), pp.717-718.
3. Dudley, S. and Simpfendorfer, C., 2006. Population status of 14 shark species caught in the protective gillnets off KwaZulu – Natal beaches, South Africa, 1978 – 2003. Marine and Freshwater Research, 57(2), p.225.
4. Bornatowski, H., Braga, R. and Vitule, J., 2014. Threats to sharks in a developing country: The need for effective simple conservation measures. Natureza & Conservação, 12(1), pp.11-18.
5. Hebert, P., Cywinska, A., Ball, S. and deWaard, J., 2003. Biological identifications through DNA barcodes. Proceedings of the Royal Society of London. Series B: Biological Sciences, 270(1512), pp.313-321.
6. Holmes, B., Steinke, D. and Ward, R., 2009. Identification of shark and ray fins using DNA barcoding. Fisheries Research, [online] 95(2-3), pp.280-288. Available at: [Accessed 25 March 2020].
7. Magnussen, J., Pikitch, E., Clarke, S., Nicholson, C., Hoelzel, A. and Shivji, M., 2007. Genetic tracking of basking shark products in international trade. Animal Conservation, 10(2), pp.199-207.
8. Clarke, S., McAllister, M., Milner-Gulland, E., Kirkwood, G., Michielsens, C., Agnew, D., Pikitch, E., Nakano, H. and Shivji, M., 2006. Global estimates of shark catches using trade records from commercial markets. Ecology Letters, 9(10), pp.1115-1126.
9. Shivji, M., Chapman, D., Pikitch, E. and Raymond, P., 2005. Genetic profiling reveals illegal international trade in fins of the great white shark, Carcharodon carcharias. Conservation Genetics, 6(6), pp.1035-1039.
10. Armani, A., Guardone, L., Castigliego, L., D’Amico, P., Messina, A., Malandra, R., Gianfaldoni, D. and Guidi, A., 2015. DNA and Mini-DNA barcoding for the identification of Porgies species (family Sparidae) of commercial interest on the international market. Food Control, 50, pp.589-596.