Planococcus sp. Isolated from sub-artic decomposing wood

Planococcus sp. Isolated from sub-artic decomposing wood.

Morgen Southwood April 26, 2017



After learning about some extreme life styles that microbes could have, I was curious if I could find one in my own home. I sampled two places, a piece of wood from my fire wood stack (which is exposed to the extreme cold temperatures of Alaskan winters) and the ashes from my stove. If any microbes grew from the firewood it would mean that they were able to withstand temperatures below -50 degrees Celsius and be psycrophiles, if any microbes grew in my wood stove they would have had to be thermophiles. I was unable to grow any microbes from the wood stove but the growth from the previously frozen wood was abundant. Each of those colonies represented a microbe that had the physiological capability of surviving those conditions, and I was interested in observing them in the lab.  The identification of this microbe would add to the depth of knowledge of what bacteria are and can be found on decomposing wood in the sub-artic.

The firewood pile was slowly decomposing, and it is likely that the microbe that was isolated from it was a part of the community that was utilizing the wood as an energy source. Decomposing communities frequently have a mix of fungi and bacteria. These communities emit extracellular enzymes to decay the wood, this results in an environment with high acidity and enzymes that are producing free radical oxygen species7. To thrive in this community it would be helpful to have mechanisms to process radical oxygen species. One such mechanism is the use of Catalase to catalyze the transformation of the reactive species hydrogen peroxide into O2. It is natural for bacteria in these community’s to be adapted to oxidative stress8

Throughout the course of this study I isolated one colony and performed many physiological and genomic tests to identify what species my isolate was. Considering the environment in which it was found, there were a few expected results: such as the microbe’s oxygen class and catalase test results. Once identified, I could then research the species and compare it’s physiology and genomic data to current literature to confirm the taxonomic assignment.



Collecting sample and isolation process

The sample was collected from a dry piece of wood. Sterile water was used to moisten the wood and a sterile swab was rolled along the wood’s surface.     The sample was streaked using the quadratic method to grow pure isolated colonies onto a tryptic soy agar (TSA) plate. To control growth rates the plates were grown in incubators for 2-4 days at 37 degrees Celsius then moved to a refrigerator to ensure colonies did not grow large enough to touch. The TSA medium provided a suitable environment for a variety of bacteria and a few molds. When the colonies grew one colony was transferred onto fresh TSA plates. The quadratic streak was used 4 times to ensure a pure culture. At this time the culture was transferred into an agar slant. Tests were performed on the isolate by obtaining cells from either the TSA plates or from the slant agar.


Physiological Characteristics

The gram stain was performed to assess the thickness of the peptidoglycan wall. The sample was stained using the provided protocol, gram-positives and negative controls were compared to the isolate to determine its gram-state (lab 4). While the cells were prepped in slides some characteristics were observed with the microscope, such as cell alignment shape and length. Colony morphology including color, size, margin and elevation was observed from growth on the TSA agar.

Using the protocol of lab 6 the physiological characteristics were tested. A suspension of the microbe in broth was tested simultaneously in all 21 of the tests included in the API 20E test strip. Additionally the isolate underwent the thioglycolate test- for oxygen class, oxidase test- for the presence of cytochrome c-oxidase, and lastly the catalase test- for the presence of catalase enzyme.

The cells inoculated onto Eosin Methylene Blue (EMB) and MacConkey Agar (MAC) plates. EMB is selective for gam-negative bacterium, if the microbe is gram negative and can ferment the sugars present then the plate experiences a color change. MAC plates are also selective for Gram-negative microbes and differentiates between lactose fermenters (agar turns pink) and non-lactose-fermenters (agar turns colorless).

The susceptibility of the isolate to 8 different antibiotics was tested using the Kirby-Bauer Method agar and the disk diffusion test per the protocol of Lab 9. The antibiotics tested were Amikacin, Cefazolin, Cefoperazone, Gentamicin, Piperacillin, Tobramycin, Trimethoprim and Vancomycin. After a two-day waiting period while the microbe was given the opportunity to develop where it could, the diameter of prohibited growth was measured and compared to standard antibiotic specific diameters and then classified as either susceptible, intermediate or resistant.



                      Dna was extracted and purified from the isolates cells using protocol from lab 5. Cell lysis was accomplished through adding sodium dodecyl sulfate (SDS), which is a surfactant that chemically weakened the cell walls, and the use of PowerBeads and the centrifuge to mechanically break the cell walls. A series of patented solutions C1-4 were added following the protocol to purify and remove inhibitors and proteins until all that remained was a pure solution of DNA. The DNA was then sent to the UAF’s DNA Core Lab sequenced using the Illumina MiSeq DNA sequencer which utilized the next-generation sequencing methodology.

Using Illumina’s BaseSpace dashboard to handle the data, the isolate’s genome was analyzed and compared to an existing database to search for similar sequences of DNA. BaseSpace has many apps that provide different information about the genomic data. The SPAdes Genome Assembler provided the overall result of the genomes assembly. Of this information, only the number of contigs that were greater than 1000 base pairs long [# contigs (>=1000bp)], The total length of the assembled genome, the longest contig and the guanine and cytosine percentage in the genome (GC%) were observed. The next app that was used was the Kraken Metagenomics, which provided taxonomic assignment. This app provided sample information such as the total number of reads that were used for classification and the percent of reads that were classified. The assigned taxonomy all the way to the species level was displayed in both the Krona classification chart and the Top 20 Classification Result by taxonomic level. Both sources have specific classification percentages that confer certainty. The final app Prokka Genome Annotation assigned gene regions that likely code for either tRNAs, rRNAs, CRISPRs and coding genes (CDs) and the abundance of them. The APP provides an extensive list of regions that were specifically identified as the above elements.

Further analysis of the genomic information was necessary. The contig.fasta file from the SPAdes Genome Assembly produced by Illumina’s BaseSpace was analyzed by BLAST. BLAST had a different database and could possibly have the genome of the isolates species if base space did not.     BLAST suggested Genus and species with associated query cover percentage and identity percentage.



Physiological Characteristics

The Isolate appeared to be a pure culture by the third quadratic streak; a fourth was performed for certainty. The Microbes transfer to the agar slant was successful: Image 1, and was successfully re-cultured from the slant when necessary.

Image 1: Microbe growing on TSA agar

Image 2: Gram-stained microbe

The gram-stain was repeated three times before a successful stain was accomplished. The gram-stain revealed that the isolate was a gram-positive coccid: Image 2. While under the microscope it was observed that the cells could be aligned linearly, in pairs or quads but not in a longer strep chain. The purity of the isolate was confirmed with the microscopic observation of a mono-culture. The cells were measured to be 1 micrometer in diameter.  Colony morphology on the TSA agar revealed a yellow colony that’s size increased with time, a smooth round margin and the colony was an elevated domed.

Image 3: API 20E test results

The API 20e tests had negative results for all for the 21 tests: Image 3. Considering that the API 20e test is designed for enteric gram negative microbes these results are confirmation that the microbe is gram-positive. The thioglycolate test had growth throughout the agar with thicker growth at the surface, which revealed that the microbe’s oxygen class was facultative aerobe. The catalase test produced bubbles and the oxidase test produced a blue color change on the strip indicating that the microbe was positive in both tests. These results reveal that the cells contained both cytochrome c-oxidase and the catalase enzyme respectively. The isolate failed to grow on EMB or MAC plates, considering that both plates are selective for gram negative, these results confirm the results of the gram positive test: Image 4.



Physiological Tests Results
Gram Positive
Cell morphology 1 micrometer diameter, coccus, cells divide linearly
Colony Morphology Yellow colonies that’s size increases with time, a smooth round margin, elevated dome.
API 20e All negative results
Fluid thyioglycolate Facultative anaerobe
Oxidase Positive result
Catalase Positive result
MAC No growth
EMB No growth

Table 1: Physiological Test Results

The microbe was susceptible to all eight antibiotics tested. The Inhibition Zones exceed the antibiotic specific threshold diameter for susceptibility see table 2 for details.

Antibiotic Name Minimum Inhibition zone for susceptibility (mm) Microbe exceed inhibition zone


Amakicin >=17 Yes
Cefazolin >=18 Yes
Cefoperazone >=21 Yes
Gentamicin >=15 Yes


    S. pneumoniae







Tobramycin >=15 Yes
Trimethoprim >=16 Yes
Vancomycin >=17 Yes

Table 2: Antibiotic Susceptibility Results


The analysis of the DNA with BaseSpace revealed unreliable data. BaseSpace’s SPAdes Gemone Assembler analyzed 128 contigs over 1000 base pairs (bp) long, a total length of over 3.7 million bp, the longest contig  was 196 thousand bp, and the GC% was 44.4%.     Kraken Metagenomics app was only able to classify 2.67% of reads, and of those reads only 17% of those reads identified the microbe as Lysinibacillus sphaericus. The Prokka Genome Annotation app identified 64 tRNAs, 0 rRNAs, 1 CRISPR, and 3739 CDS.     Image 5 and 6 are from the Kraken app.

Image 5: Krona Classification Chart from BaseSpace displays the large portion of unclassified reads.

Image 6 To 20 Classification Results by Taxonomic Level from BaseSpace

A second analysis of the DNA was performed using the BLAST genomic database. BLAST classified the organism as Planoccus donghaensis with a query cover of 77% and a 78% identity. Several other species had similar query cover, including Planococcus antarticus: Image 7.

Image 7: BLAST Genomic Results


The genotypic test results for this microbe were vague, the results of BaseSpace had a low number of reads that could be classified and of those reads there was little consensus. The results of the SPAdes Genome Assembler were above required values; # contigs >=1000 bp should be in the hundreds, total length should be at least a million bp and the longest contig should be at least 100000 bp. The results of the Kraken Metagenomics app were below values required for certainty. The percent of contigs read needed to be greater than 80% and the percent of those reads that needed to agree upon a classification to trust that classification to the genus level was 80% and to the genus level was 60%. The results did not meet these thresholds, and this is the reason that the BaseSpace indentified taxanomy of Lysinibacillus sphaericus was rejected.

BLAST provided many species level classifications with similar query cover and percent identity. The species classification with the highest query cover is poorly documented in literature. The first publication of a species under the genus Planococcus was P. citreus. It was identified in 1894, and was only approved in 19806. This genus is relatively young and is currently growing with many of the species being described within the last ten years. Of the few documents that relate to Planococcus donhaensis, there is one that notes the origin of the sample, the South Korean Sea. There are some qualities that are consistent between my sample and P. donghaensis, they are both, gram-positive, aerobic, coccus and oxidase positive 1. However these similarities are not enough to convince me that they are the same species. A hallmark of the genus Planococcus is that they are usually halo-tolerant gram-positive bacteria that frequently inhabit Antartica4. The isolates holotolerance was not tested but the similar cellular membrane composition and habitat conditions indicate that this genus is likely correct.

The negative results and lack of growth in the API 20e, MAC, and EMB are all consistent with the microbe’s gram-positive cell wall. These tests reveal nothing more than a conformation that it is indeed a gram-positive microbe. The oxygen class determined by the fluid thioglycolate test and the positive results of the Oxidase and Catalase test are both consistent with the oxygenic environment that the microbe was isolated from. These tests proved that the microbe could thrive in an oxygenated as well as an anoxic environment, that it contained cytochrome c oxidase which is a part of the electron transport chain found in microbes that utilize oxygen, and that it contained the catalase mechanism for dealing with oxidative damage, respectively.

At the genus level, morphology can vary greatly. Many of the morphological and physiological analyses made on the isolate are not held by every species in the Planoccocus genus. The morphological characteristics that are consistent with the isolate and across the genus are: gram-positive membranes, cocci cell shape, colonies are yellow orange in color, catalase positive and an aerobic oxygen class5. My isolate was not just aerobic but a facultative aerobe, which is inconsistent with two articles that state the genus is strictly aerobic5. The isolate was found to be susceptible to all antibiotics tested with the disk diffusion test. This is consistent with a study that found, to the level of the genus, that Planococcus’ were susceptible to all antibiotics tested3.

The second most likely species suggested by was P. antarticus, at least this species shares a similar environment. P. antarticus thrived in an Antarctica, my sample would likely have similar mechanisms for surviving temperatures around -45 degrees Celsius in Faribanks winters. P. donghaensis may have had the ability to deal with these temperatures in its genome, but it is not certain since it’s environment doesn’t select for such characteristics.

The literature on the Planococcus species like donghaensis, kocurii, halocryophilus and antarticus often notes how closely related the species are and how further analysis like G-C content, fatty acid strains, DNA-DNA hybridization etc1,2 are needed to differentiate the species. To conclusively Identify the taxonomy of this isolated microbe, it would be advisable to repeat DNA isolation and genomic analysis that was preformed in this project, and to additionally perform other genotypic analyses like, DNA-DNA hybridization and 16s rRNA analysis and fatty acid identity. Considering the limit of species identified to this date, there is the potential that this microbe could be a new species.




Work Cited

  1. Jeong-Hwa C, Wan-Taek I, Qing-Mei L, Jae-Soo Y, Jae-Ho S, Sung-Keun R, Dong-Hyun R. Planococcus donghaensis nov, a starch-degrading bacterium isolated from the East Sea, South Korea. Int J Syst Evol Microbiol. 2007;57:2645—2650. doi: 10.1099/ijs.0.65036-0.


  1. Reddy G, Prakash J, Vairamani M, Prabhakar S, Matsumoto G. Shivaji S. Planococcus antarcticus and Planococcus psychrophilus nov. isolated from cyanobacterial mat samples collected from ponds in Antarctica. Extremophiles. 2002;6:253—261.


  1. Tuncer, I. (2016). Antibiotic resistance of bacterial isolates from sediments of eastern Mediterranean Sea in association with environmental parameters. Journal of Bacteriology and Parasitology, 7(6), 51.


  1. See-Too, W. S., Ee, R., Lim, Y.-L., Convey, P., Pearce, D. A., Yin, W.-F., & Chan, K.-G. (2017). AidP, a novel N-Acyl homoserine lactonase gene from Antarctic Planococcus Scientific Reports, 7, 42968.


5.         Fackrell, H. (n.d.). Planococcus. Retrieved April 11, 2017, from website:

  1. Euzeby, J. P., & Parte, A. C. (2017, April 1). Genus planococcus. Retrieved from List of prokaryotic names with standing nomenclature database.


  1. Valášková V., de Boer W., Gunnewiek P. J. K., Pospíšek M., Baldrian P. (2009).  Phylogenetic composition and properties of bacteria coexisting with the fungus  Hypholoma fascicularein decaying wood.  ISME J.  3  1218—1221. 10.1038/ismej.2009.64
  2. de Boer W., van der Wal A. (2008).  “Interactions between saprotrophic basidiomycetes and bacteria,’ in  British Mycological Society Symposia Series 8  eds Lynne Boddy J. C. F., van Pieter W., editors. (Cambridge, MA: Academic Press; )  28  143—153.





Edible Microbed

My art is entirely edible, though there may be some of you out there that hate fondant as much as me, and would not consider it very edible…

My cupcakes are all decorated with microbe related items, some display  different morphologies, like coccus or bacillus, or different features like flagellum or slime.  I got my inspiration by looking for microbial cupcakes online, there were some real masterpieces out there, and they made it look easy.  My cupcakes turned out to be less than masterpieces, the icing I used to do things like the T-4 bacteriophage, and the super coiled DNA melted… in the FREEZER.  I definitely wont use that recipe again. My finished project may not be as attractive as I had hoped, but it is just as tasty.

This first set of microbes was taken before  they melted.  I will probably make more cupcakes closer to the potluck so that they wont have a chance to melt

A2: Microbes in the News

Article title: Scientists turn food poisoning microbe into powerful cancer fighter

Date: Feb 8, 2017

Author :Michael Price

Source: Science



Assignment author: Morgen Southwood



Cancer cells are not attacked by our immune system because they are considered to be ‘self.’ There has been some successful research into marking cancer cells for immune system attacks through the use of bacteria. One such treatment for bladder cancer is already approved by the FDA. Bacteria can target cancer cells because they tend to be necrotic and oxygen depleted, which is the kind of weekend tissue pathogens naturally target.     This article focuses on a study on the use of Salmonella. In mouse studies it has been found to eliminate/reduce tumors, and even help prevent secondary tumor formation from human colon cancer metastases injected into mice. The Salmonella used in the studies was altered to produce FlaB which is the protein the spurs the elevated immune responses.


This article strongly related to the lectures we are having on immunity. One element of the lectures that is particularly relevant is the way that the immune system has an amplified response the second time it encounters an antigen, and the specificity of antigen identification. From what I understand, this treatment would only deal with tumors that are present as long as the altered Salmonella is present. Since the Immune system is attacking Salmonella, and hasn’t been taught that the cancer cells were the danger all along, this treatment can not cure or prevent future cancers.

Critical analysis-

I really enjoyed the level of detail that this article went into, it described relevant methods and results in a clear and concise manner. I think the terminology used, the explanations provided made this article accessible and informative for the public. What I learned from this article is the targeting mechanism involved in bacterial cancer treatment. I find it very interesting that bacteria can be so relied upon to target the necrotic oxygen depleted cancer cells, and not healthy ones.  One thing I noticed in this treatment is that it has a narrow application. This is not the cure for cancer, it is only one more weapon. I do not this this treatment could be used on an immune compromised person, or on cancers that have morphological of physiological characteristics that cant be targeted by bacteria.


They have performed an experiment where they injected colon cancer cells into 20 mice. They used the treatment on half of them and observed them for 120 days to see if the cancer remained. In the next experiment they injected metastasizing colon cancer cells into six mice and after 27 days the six treatment mice and 7 control mice were examined for tumors. Briefly, describe what the next experiment on this treatment should entail.

A2: Microbes in the news

Article title: Behind the iron curtain: How the methan-making microbes kept early Earth warm Behind the iron curtain: How methane-making microbes kept the early Earth warm


Date: Arpil 17, 2017

Article author was not listed but the research’s author was: m S. Bray. The article was provided by the Georgia institute of Technology.



Assignment author: Morgen Southwood


Marcy Bray and his team simulated early earth conditions to try and explain why the oceans could be liquid in the first two billion years. The prevailing theory is that methanogens provided enough methane for the green house effect to maintain liquid oceans. The problem with this theory is that, as we learned in class, methanogenesis is an inefficient system, and can be out competed when alternatives are possible. One major competitor in this time period was the rust-breathing microbes, they would dominate any environment when iron was available. The term iron curtain, refers to the potential for rust-breathing microbes to repress methane emissions when rust is plentiful. If methane was completely suppressed then the planet would likely have cooled. The microbiologists simulated early earth to study microbial diversity and methane emissions in varying conditions. They found that in iron free pockets of the oceans, methanogens could have thrived and been enough of a source of methane for keeping early Earth warm.


This article strongly related to our lectures on the methane cycle It also related to some exam 1 material, when we learned about the ferrous and ferric iron signatures that signaled changes in early earth microbial diversity.

Critical analysis-

This article could have used some more explanations. I understood the conclusions it drew, but I wouldn’t have been able to without material I learned in this class. I would have needed someone to fill in the blanks for me. It was important to understand why rust-breathing microbes would have outcompeted methanogens, and the significance of shifts in microbial diversity with different conditions etc. The article assumed/required the reader to know this supplementary information, and therefore it was not accessible to the general public.  I think the article was scientifically accurate in the way it described the idea proposed by the results of the study, however the title is misleading. The title seems portray that the study was a confirmation, when it was only supportive of the idea.

Since I did have some background information, this article was very interesting to me. When I thought about the major shifts in microbial diversity of the planet , I always thought about microbes relating to oxygen. These rust breathing microbes and methanogens were just as important stepping stones in shaping the Earth.


The conclusion of the article is that methane emissions could have come from microbial communities that were in rust free patches of the ocean. I thought that the ocean was well mixed. How could there be sections of the early ocean that were so poorly mixed that they lacked iron, while other areas had high levels of iron?


A2: Microbes in the news

Article Title: Deepest Life on Earth May Be Lurking 6 Miles Beneath Ocean Floor

Author: Thea Ghose

Date: April 11, 2017

Source: Live Science


Assignment author: Morgen Southwood


There are mud volcanoes under the sea floor and they may be inhabited. Biological signatures in material that has risen to the surface could possibly be coming from microbial life 32,800 feet under the surface of the ocean floor. The organic matter are good indicators of the presence of life, but could also have been produced by abiotic processes.


Reading this article reminded me of earlier in the semester when we discussed the origins of life and more recently in the semester when we discussed Bas-Becking’s idea. When we discussed the origins of life near under water vents we discussed the kinds of organic chemicals that could have been precursors for the first life forms, those same compounds were found in these mud deposits. When we discussed that life could/would be everywhere, I considered the presence of life beyond our atmosphere, but not beneath the crust of the earth. This article made me wonder what microbial super power could survive in those conditions.

Critical analysis. —

This article only spends one half of it’s very short article discussing the new discovery. I wish that there had been more details on the discoveries methods. Apparently the compounds came from rocks that were “spewed’ onto the surface, there was no explanation on how the scientists could be sure that any signs of life originated in the mud volcano, and wasn’t the product of contamination as the mud progressed to the surface.

My favorite sentence in this paper isn’t referring to the recent discovery; it’s within a paragraph summarizing other research on deep sub surface microbial life. The sentence reads, “ the deeper that scientists have looked the deeper life has seemed to go.’     I wonder if there is a limit to this, if scientist will one day conclusively say: no more life past this point. The researchers of the under sea mud volcano seem to think so. They made an estimate for the maximum depth that could support life. Considering a maximum temperature of 122 degrees Celsius and 1000x atmospheric pressure, the deepest Achaean environment would be about 32,800 feet below the surface.

This article was presented both scientifically, and in a way that could be digested by an interested member of the general public. Its lack of depth was compensated by links to relevant background information and relevant studies.


Will scientists ever be able to definitively state that an environment/ location is completely free of life without having to clarify “that we know of’?


Morgen Southwood Microbial Worlds Assignment


Artist: Sarah Tabbert

Piece, Printmaking and woodcarving

The pieces are inspired by photos taken with the magnification of a microscope. Some of them are her photos and some are not. I found the images visually appealing; I liked the use of color to liven the microscopic images. What made the pieces most attractive to me was her connection of the sensation of looking through a microscope and goings snorkeling. I hadn’t ever considered that before, but once she mentioned it I realized that I felt the same way.



Artist: Nancy Hausle-Johnson

Piece: Emergence: The waroming Climate is waking up sleeping microbes.

This piece is probably my favorite. The vibrant photos demand attention. The three panels are from different stages of thawing permafrost. Permafrost provides the ability to travel through time to the point at which these microbes were preserved. Many of the microbes can be revived, which means that with thawing permafrost, there is be a massive immigration of time traveling microbes that will have the potential to alter the fluxes and sinks of nutrients and chemicals in our artic and global environment. This analysis of microbes at different stages of the thawing process can give estimates on what the future ecology of artic microbes will be with a changing artic. This piece is not only beautiful, but a fascinating peak at the future and the past. I wouldn’t change a thing about this exhibit.



Artist: Ree Nancarrow

Piece: Deceptive Beauty

This piece uses the quilting as the medium to show several elements relating to methane in the artic. The central column sows bubbles rising from what I assume is an anoxic population of methanogens. The methane swirls in the central pane, and is burned in the top (which may represent methane literally burning or the warming greenhouse effect methane has). There are panels on the left and right that look like methane bubbles in liquid water, and some that look like truly realistic frozen bubbles. I really enjoy the top panels of the black spruce, it makes this piece feel like its locally relevant, not just a depiction of any old methane source, but an Alaskan one.


4) If I was going to present an art piece at this exhibit I think I would have worked with something related to the human micro biome. I really like the image that is associated with Dr. Drown’s micro biome class. Maybe I could have isolated really microbes from my body and then collaged them together to make a model human.

Eric Collins extra credit

Eric Collins discussed aquatic samples that were gathered near artic ice and the micro biome that was within it. He discussed the origins of the water and the chemicals in the artic ocean, what land masses they melted from, what current brought the water there and took water away. Collings discussed the affect that melted sea ice has a chemical finerprint on the water it melts into, and how that relates to the micro biome in the artic. He also examined the ways that artic climate change could cause microbial extinctions.


He mentioned that the climate change in artic oceans could cause microbial extinctions that couldn’t/wouldn’t happen in other biomes. This made me think about a quote we heard in lecture, about how microbes are everywhere the environment selects. I think that this quote doesn’t mean that every species will be present everywhere just that if it could be there it would be. Back to the melting artic, any microbe that requires a saprophytic environment, and or the specific chemical composition of water associated with melted sea ice will loose its niche if the ice melted. These microbes have no where to go, they are already at an extreme environment, that simply wont exist in that future. Microbes in other environments will be able to migrate across latitudes and altitudes, but not these microbes, they wont have anywhere to be. It’s a bit sad to think about.


Morgen Southwood

painting with microbes

Morgen Southwood

I don’t have much in the way of artistic skill, I thought that doing some swirls would be fun in my other plates but they ended up quite ugly, so I’m only showing my TSB. Considering that the microbes were all displayed on tsb plates it was very easy to know what the colors the colonies’ pigments would produce. My image is supposed to be an artsy take on the basic idea of a cell, The membrane is pretty pink and wavy encircling the cell. The nucleus is unrealistically shaped like a flower (but circles are boring) There are swirls and swoops about the central flower, they are kind of like the ER. I like the happy accident of my lighter white streaks getting some of the bolder white colonies to grow in them- it’s kind of like rough ER now.