The Skin of The Sea: where the sky touches the Ocean

Everything in nature serves a purpose, even if we do not understand it or fail to identify it. Some natural processes are so subtle it can be easy to think that they simply do not exist. Look at the sky and then at the ocean: one over the other, so distinguishable, and never seeming to touch. But they do. There is a thin layer of ocean surface in contact with the air, and in it, many chemical and biological processes take place.”  schmidtocean.org

A culture of the bioluminescent bacterium Photobacterium phosphoreum in a tea cup.The complex fluctuations in bioluminescence are generated by the bacteria as they respond to the highly dynamic film that is the Sea Surface Microlayer. 

Exploring the Invisible 2009-2011 was a Wellcome Trust funded collaborative project between artist Anne Brodie, myself, and writer and researcher Caterina Albano. The work explored the bioluminescent bacterium, Photobacterium phosphoreum, a light-emitting marine lifeform commonly found in sea water.

Through enquiry and experimentation, that transcended the traditional boundaries of art and science, the project developed a large body of photographic and moving image works and a number of live installations that reimagined our encounter with these beguiling light emitting bacteria. Please follow the link below for more information on the project

http://www.annebrodie.com/#/exploringtheinvisible/

Anne and I would spend many hours in the dark room interacting with these bacteria,  and under the influence of their beguiling cold blue light. When we were working with artificial seawater liquid cultures of them, we noticed that when we had turned our backs on the large culture flasks, and allowed them to become still for a time, the bioluminescence began to form astonishing and complex patterns at the air/liquid interface. We documented this process in a number of works. Please see the videos below.

A culture of the bioluminescent bacterium Photobacterium phosphoreum in a Petri dish. The complex fluctuations in bioluminescence are generated by the bacteria as they respond to the highly dynamic film that is the Sea Surface Microlayer.

A culture of the bioluminescent bacterium Photobacterium phosphoreum in a gravy boat. The complex fluctuations in bioluminescence are generated by the bacteria as they respond to the highly dynamic film that is the Sea Surface Microlayer.

 

A culture of the bioluminescent bacterium Photobacterium phosphoreum in a wine glass. The complex fluctuations in bioluminescence are generated by the bacteria as they respond to the highly dynamic film that is the Sea Surface Microlayer.

We didn’t know it then, but what we were observing was the activity of the sea surface microlayer (SML) or the skin of the sea. This microlayer is the top 1000 micrometres of the ocean surface, and the boundary layer where all exchange occurs between the atmosphere and the ocean. This “skin” develops at the surface of the ocean (and also on lakes and ponds) where organic compounds come into contact with the atmosphere. It affects how quickly gasses can exchange between the atmosphere and the ocean, and is critical in carbon dioxide exchanges and climate change modelling. Moreover, much research has shown that the SML contains elevated concentrations of bacteria, viruses, toxic metals and organic anthropogenic  pollutants as compared to the sub-surface water. The complex fluctuations in bioluminescence are generated by the bacteria responding to this highly dynamic layer.

In the videos above the phenomenon is viewed from above. Below is a video of a small cell with the activity of the SML being viewed side on.

The Ubiquity of Possibility

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A purple microbial ecology on the glass roof of Guildford Station

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A purple microbial ecology on the glass roof of Guildford Station

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A purple microbial ecology on the glass roof of Guildford Station

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A purple microbial ecology on the glass roof of Guildford Station

Glass is an anthropogenic material that is amongst the most inimical to microbial growth. Yet, here on the glass roofs of Guildford Station a thriving microbial  ecology has managed to establish itself (images above).  I’m not sure what it is but it looks  like the photosynthetic purple sulphur bacteria that you would usually find illuminated anoxic zones of lakes and other aquatic habitats or in Winogradsky columns. It’s as if the air fizzes with all manner of latent biological potential, in the form of its bacteria, that are just waiting to find an appropriate niche in which to thrive. Whatever this roof top ecology is made up from, it must be an exquisitely matched to the environmental niches because just a couple of yards away on the other side of the platform there is a very different one (images below). A fine example of my concept of microgeography.

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On the other side of the platform, just a few yards away, a very different microbial ecology

BactoGrams

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BactoGrams. Made by differentially exposing bacteria to short wavelength UV light. Those exposed die (too many thymine dimers in their DNA to repair), those shielded grow. Not as sophisticated as those by Zachary Copfer yet but getting there! Made with red, yellow, pigmented,  and bioluminescent,  bacteria.

Clot Magazine

” Park’s explorations and research intend to put out there the hidden wonders of bacteria for the public to enjoy and be delighted with. He blends art and microbiology with such beauty and meaning that some of us would have never guessed possible. A lovely and eloquent write up of my purpose and work here.

Article here

Thank you Clot Magazine!

 

 

For Christmas, a bacterial love story: The Incredible Journey

Figure 1. The Cyanophyte Oscillatoria animalis growing on agar (left) and viewed under a low-power  microscope (right)

I’m exploring the use of the photosynthetic and auxotrophic Cyanophyte (Cyanobacterium) Oscillatoria animalis  (Fig. 1) as novel BioMaterial that can be fashioned from little more that sunlight, water and air.

When this filamentous bacterium grows in shallow culture media it self-organises into a floating bladders (Fig.2) that are reminiscent of silk moth cocoons. In fact, the bladders here are also woven from millions of microscopic filaments.

Figure 2. Floating bladders of Oscillatoria animalis, each one is woven  millions of microscopic filaments

I discovered the remarkable properties of the structures formed by Oscillatoria  when I accidentally knocked a cultured onto the floor. Fortunately, the bottle was plastic and so didn’t break, but my Oscillatoria structure that had taken 3 weeks to grow  had turned into a green soup. However, when I returned from a coffee break about 15 minutes later the Cyanophyte structure had miraculously repaired itself. The video below is a recreation of the accident, but here I deliberately shook the culture. The time-lapse represents 10 minutes of real time

 

Now to the love story. To grow the Oscillatoria structures, I clone them, that is I cut them into two, and transfer each cut-half into a new culture dish, where they grow into larger bladders again. The process is then repeated again and again. On one occasion that I did this I’d run out of fresh media so I left the two freshly separated halves to separate and float apart across the distance of the Petri dish. The uncut Oscillatoria bladder can be seen in Fig 3. below.

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Figure 3.The uncut Oscillatoria bladder

The image (Fig 4) below shows the cut bladder and freshly separated halves.

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Figure 4. The two newly split daughter forms of the Oscillatoria bladder

 

Now this is where the love story really begins. When I returned the lab, the following morning, after having cruelly separated a union, the two halves had reached out to touch each other across the distance of the culture dish (Fig 5). For the microscopic cells of Oscillatoria,  the distance over which this re-uniting grasp occurred, must be the equivalent of  reaching out over massive planetary  distances for us. You can see this first recontact in the image below (Fig 5)

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Figure 5. Reconnection of the two Oscillatoria  forms across 10 cm of liquid culture

 

Over the next few days and in the images that follow  (Figs 6-9), the two separated halve slowly pulled each other until over vast distance for them, they were finally reunited.

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Figure 6 . The two Oscillatoria forms slowly pulling themselves together fivesix

Figures 8-9. The two Oscillatoria forms slowly pulling themselves together

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Figure 9: Finally the two Oscillatoria structures fuse and reunite having once been separated by vast distances. The end of an Incredible Bacterial Journey 

Skin Flora:the accidental pathogen, the golden globe, and pink swans

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Staphylococcus epidermidis (left), Kocuria rhizophila (middle) and Kocuria rosea (right)

 

Here are some new additions to C-MOULD, a unique culture collection specifically for microorganisms for use in art and design. 

Staphylococcus epidermidis: the accidental pathogen (left) is a very close friend of Staphylococcus aureus and MRSA. Whilst it isn’t as pathogenic as these bacteria has is a very important opportunistic pathogen, not able to cause illness in otherwise healthy people, but capable of infecting the compromised human host. In fact, it is now the most frequent cause of nosocomial infections, at a rate almost as high as that due S. aureus, its more virulent cousin Staphylococcus.  It also the most frequently isolated bacterium from human skin and is especially common in damp areas like the groin and arm pits. Skin is a surprisingly harsh environment for bacteria, and analysis of the S. epidermidis genome shows that this bacterium is well equipped with mechanisms that protect it here, and especially, allow it to tolerate extremes of salt concentration in the form of dried human sweat.

Kocuria rhizophila, formerly known as Micrococcus luteus (middle): sunscreens and the golden globe is also another common human skin inhabitant and is another bacterium that has adapted to be able to survive in this harsh environment. Like human skin is, bacteria are susceptible to the damaging effects of Ultraviolet Light (UV) and so K. rhizophilia synthesises a pigment that absorbs wavelengths of light from 350 to 475 nm. These wavelengths of UV, commonly referred to as UVA, have been correlated with an increased incidence of skin cancer, and it is possible that this pigment could be used to make a sunscreen that can protect humans against UVA. K. rhizophilia is also an important bacterium in the context of the history of microbiology and medicine, as it played a key role in Alexander Fleming’s discovery of lysozyme, to which it shows exquisite sensitivity (Fleming, 1922a,b). In fact, it is readily killed by the lysozyme present in human tears irrespective whether they are generated by happiness or grief. Finally, variants of K. rhizophila can precipitate gold by concentrating and crystallizing it on their surface.

Kocuria rosea: fake tans and pink swans: again is a commonly isolated bacterium from human skin. Its adaptation to this environment is its ability to breakdown and thus utilise keratin, the key structural protein making up the outer layer of human skin. The name of the species rosea, gives a clue to the colour of its colonies which are pink. It also produces a variety of different pigments one of which is canthaxanthin. This is a rust coloured pigment that is approved as a food additive in a number of countries. It is also the active component of tanning pills. When taken at these large doses, many times greater than the amount normally ingested in food, this substance is deposited in various parts of the body, including the skin, where it imparts a golden orange hue, because it accumulates in the  panniculus. However, when consumed in these high doses it causes canthaxanthin retinopathy, which can lead to loss of vision, because the pigment accumulates in the macular (the central part of the retina). Finally, by virtue of its colour and keratin digesting activity K. rosea may also be involved in a mysterious condition in swans called Pink Feather Syndrome, in which the birds develop a pink coloration on their feathers.  Overtime, the pink feathers become degraded, and the swans may die.

 

 

 

 

 

 

 

 

 

 

 

pDENIM: towards a sustainable denim grown entirely from bacteria

 

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A sheet of bacterial nano cellulose observed under the microscope, 200x magnification

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A sheet of bacterial nano cellulose observed under the microscope, 200x magnification

The over arching aim of the pDENIM project (porkayoticDENIM) is to make a sustainable form of denim that is made and grown (both textile and dye), entirely from bacteria.

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An agar plate with a culture of the hyper cellulose producing bacterium GXCELL

The textile for pDENIM will be made using GXCELL a specialised hyper-nanocellulose producing bacterial strain isolated from a kombucha scoby. When this bacterium is grown in a liquid  mixture of vegetable extracts,  it forms mat of cellulose that are far thicker than many other strains of cellulose producing bacteria (see figures below)

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A comparison of cellulose production by GXCELL (left) and a normal cellulose producing bacterium (right). After 5 days incubation

 

 

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A disc of bacterial cellulose produced by GXCELL

 

When the bacterial cellulose is dried it forms a thin and plastic-like film, which is brittle (see below) and clearly unsuitable for use as a  flexible textile such as denim.

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The dried form of bacterial cellulose

 

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The dried form of bacterial cellulose

 

However, I have recently developed a process that converts the dried form of cellulose into a highly flexible textile, somewhere between paper and cotton (see below to see a refolded swatch of this material), and which will now become the bacterial fabric component pDENIM.

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The new flexible form of bacterial nanocellulose

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The new flexible form of bacterial nanocellulose

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The new flexible form of bacterial nanocellulose

 

The other avenue of investigation for the pDENIM project seeks to find a sustainable and bacterially derived pigment to replace indigo, the dye used to stain denim blue. This dye  was traditionally extracted from plants of the genus Indigofera. Today, however, the several thousand tons of indigo used each year is synthetic and is produced by industrial processes with obvious consequences for the environment. Whilst many bacteria are pigmented  (e.g. red, purple, yellow and pink), those that produce strong blue pigments are rare. The first blue pigment bacterium that I investigated was Vogesella indigofera, a rare blue naturally pigmented bacterium that was originally isolated from a pond that had been used as a dump for highly toxic chemical waste (see below).

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A single colony of the blue pigmented bacterium Vogesella indigofera.

 

I find it intriguing that a life form so beautiful could arise from such a polluted environment. However, despite its beauty this bacterium was not a suitable replacement for indigo because its blue colour was too faint.

The second blue-pigmented bacterium that I characterised was Arthrobacter polychromogenes, which produces the water-soluble blue pigment, indochrome, together with the insoluble blue pigment, indigoidine (see below).

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Colonies of the blue pigmented bacterium Arthrobacter polychromogenes.

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Colonies of the blue pigmented bacterium Arthrobacter polychromogenes.

 

In the images below Arthrobacter polychromogenes has been grown on cotton and dyes it a deep blue. The next step will be mix the bacterially grown textile with the bacterially generated blue dye to make pDENIM. Watch this space……………..

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A swatch of cotton dyed blue by the growth of the bacterium Arthrobacter polychromogenes.

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A swatch of cotton dyed blue by the growth of the bacterium Arthrobacter polychromogenes.

 

Mutator: a unique radiation detecting BioInk

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Bacteria are essentially haploid, that is they only contain one copy of their genome. Consequently, unlike higher organisms which are usually diploid, and which containing two copies of the genome, changes in the DNA sequence of the genome (mutations) usually result directly in measurable phenotypes. There is no blending or filtering of the genome’s informational content as is the case with higher organisms and mutations or glitches in the genetic code can become readily visible.

The bacterium Serratia marcescens usually grows as a distinctive and red pigmented colony. However, if it is exposed to agents which cause DNA mutation, and these genetic changes occur in pathway that gives rise to the red pigment, then the bacterium will become white.

In this process here, I have dramatically increased the natural mutation rate of the genome of this bacterium by exposing it to radiation. The damage to its DNA can clearly been seen in the emergence of many non-pigmented white mutants, amongst the red-pigment parent strain.

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It strikes me that this system could be used as a living biological ink which would record exposure to radiation, and perhaps form the basis of a biological radiation detector. Maybe it’s already sniffing out and detecting the legacies of Chernobyl and Fukushima.

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Resistant Materials: an aesthetic exploration of antibiotics and antimicrobial resistance

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The disc diffusion test is a commonly used, and important, method for determining antibiotic sensitivity of bacteria in the clinical and research laboratory.  In this test, paper discs impregnated with different antibiotics  are placed onto an agar plate in which bacteria have been spread all over the surface, and the culture is then incubated. If an antibiotic is effective against a particular antibiotic, it will prevent the bacteria from growing, or kill them, so that there will be an area around the disc  where the bacteria have not grown. This is called a zone of inhibition, and its appearance indicates sensitivity of the bacterium to the antibiotic. When a zone of inhibition does not appear, or is very small, then the bacterium is resistant to the antibiotic.

To make this aesthetic interpretation of the disc diffusion assay, I chose to use two naturally pigmented bacteria, Serratia marcescens (Sm), red,  and Chromobacterium violaceum (Cv), purple and to test their sensitivity to three antibiotics, namely Cloxacillin (C), Kanamycin (K) and Trimethoprim (T). Moreover, instead of using the conventional circular discs for the assay and chose to use antibiotic impregnated papers that had been cut into letters.

In the first run of this process,  the sensitivity of Sm and Cv to C and were investigated. After 12 hours incubation it became apparent that CV was resistant C, as it grew right up to the antibiotic impregnated paper, and in fact,  even grew though it.  On the other hand Sm was sensitive to C, as evidenced by the large and clear zone of inhibition (see below).

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Sm left and Cv right. The reversed L shape on the lefthand side of the agar plates is impregnated with K, and the L on the right with C.

 

After 16 hours of incubation, mutant  colonies of Sm that were resistant to C had begun to emerge. A direct visualisation of evolution and the emergence of  antimicrobial resistance (see below).

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A close up photograph of the zone inhibition in Sm caused by C reveals small red C resistant mutants.

 

After 24 hours incubation the C resistant mutants of Sm are clearly visible but neither bacterial strain, Sm or Cv, displays any resistance to K (see below).

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Sm left and Cv right. The reversed L shape on the lefthand side of the agar plates is impregnated with K, and the L on the right with C.

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A close up image of the C resistant mutants of Sm

 

In the second run of this process, the same bacteria were used, but the antibiotic Trimethoprim (T) replaced C and K. In the image below is the first run of this process (top) and run with T (bottom) but before incubation.

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Top, first run of the process. Bottom, second run, left Sm with T, and bottom right Cv with T.

After 12 hours of incubation both Sm and Cv exhibit sensitivity to T, as evidenced by large and clear zones of inhibition(see below).

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Top, first run of the process. Bottom, second run, left Sm with T, and bottom right Cv with T. Note also how the fungal colonies top left are resistant to both C and  K. 

 

In a very unexpected development, after 24 hours of incubation Sm, but not Cv, had become resistant to T. However, Sm in doing so,  had lost its ability to produce its red pigment and thus become white in the process (see below).

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Left Sm with T, and right Cv with T. The T resistant form of Sm is white. There might also be a white T resistant form of Cv emerging at the bottom of the T.

The Animalcule Drawings I: Black Carbon (BC)

” Unique conflicting BioArt made with life and Anthropogenic Materials”

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Antonie van Leeuwenhoek is widely regarded as one of the founding father’s of microbiology, as he was one of the first individuals to observe protozoa (1674) using a microscope that he had made himself. In a letter written in September, 1674, Leeuwenhoek describes a spiral microorganism, possibly what we know of today as Spirogyra, in his observations on lake water,

“Passing just lately over this lake, . . . and examining this water next day, I found floating therein divers earthy particles, and some green streaks, spirally wound serpent-wise, and orderly arranged, after the manner of the copper or tin worms, which distillers use to cool their liquors as they distil over”

Van Leeuwenhoek originally referred to these microscopic creatures as Animalcules (from the Latin animalculum, meaning tiny animal). Such microscopic life forms are also called  Infusoria from the Latin infundere (infuse) and so named because they were originally found in infusions of decaying organic matter. Today, these micro-organisms referred to as unicellular organisms or Protozoa (meaning first animals). As the principal hunters and grazers of the microbial world, protozoa play a key role in maintaining the balance of bacterial, algal, and other microbial life in natural and urban ecosystems. They also are themselves, an important food source for larger creatures and form the basis of many food chains. Protozoa have been found in almost every kind of soil environment, from peat bogs to arid desert sands, and vast numbers microscopically animate, the deep seas,  near the surface waters, form the tropic to  frigid Arctic and Antarctic waters.

Protozoa, have evolved many different strategies for feeding some are sessile and acquire nutrition through filter-feeding,  whilst others are motile and actively seek out their prey. In both, cases these activities are powered by organelles called cilia or flagella which beat  wave, allowing which allow the protozoa to move and to generate powerful micro-currents, to filter feed.

Humans pollute the environment with huge quantities of soot every year and it is believed  to be the second most important human-made agent of climate change. It is generated by burning of forests and savannah, by diesel engines, and coal burning. A major constituent of soot, is Black Carbon (BC) which can absorb one million times more energy than CO2. BC is the second largest contributor to climate change after CO2. However, unlike CO2, which can stay in the atmosphere for thousands of years, BC, because it is particulate, remains in the atmosphere only for days to weeks before it returns to earth with rain or snow. When it falls to earth with this precipitation, not only does it damage ecosystems and reduce agricultural productivity, but BC also darkens the surface of snow and ice, reducing their albedos (the reflecting power of a surface), warming the ice, and hastening melting.

Air pollution also  causes millions of premature deaths each year, these being mainly caused by the inhalation of microscopic particulate matter. BC is especially dangerous to human health because of its tiny size. In fact, the WHO reported in 2012,  that there was sufficient evidence to link exposure to BC to cardiopulmonary morbidity and mortality.  The review of the toxicological studies suggested that BC may not be a major directly toxic component of fine particulate matter, but that it may operate as a universal carrier for a wide variety of toxic chemicals into the human body.

India ink, commonly used in drawing and in art, has an intriguing link with Black Carbon (BC) in that a form of BC called Lampblack is its main constituent, other than water.  Usually no binder material is necessary in order to stabilise the ink as the BC particles are in colloidal suspension, and they form a waterproof layer after drying. Lampblack is made from burning oily or resinous materials and collecting the resulting soot containing the BC.

To make the Animalcule Drawings, I mixed various animalcules with India Ink and observed them under a Differential Interference Contrast microscope at 200x magnification. This  processes simultaneously reveals the complexity of these lifeforms and the complex fluid dynamics that their activity generates, and also the insidious nature through which a widespread and anthropogenically generated pollutant, Black Carbon,  infiltrates living organisms, and the enters into the bottom of the food chain.

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Blepharisma

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Unidentified fast moving animalcules

 

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