Agar Art

With the conclusion of this semester, here is some additional pieces I have created! With all of the species I have in my collection, I decided to do one last push for creative art. I have learned a lot about bacteria in this time, including different growth rates, growth environments, how to plan, organize and perfectly time planting and harvesting of colonies, and refined my aseptic technique and fine motor skills. This journey has taught me a lot about patience as well, as bacteria and myself can only grow so fast… But now let’s get back to the science!

Using E. coli (White), Serratia marcescens (Red) and Rhodotorula glutinis (Pink), I created the classic “Cupid’s Arrow” art piece.

Using Pseudomonas aeruginosa I have created a four-leaf clover, representing good luck to whoever reads this.

The Spring/Easter line up features many different species, some of which I do not know. Some of the species I have used however are E. coli (White), Rhodotorula glutinis (Pink), and Staphylococcus aureus (Yellow-Gold). Here I have created an Easter Egg, the Easter Bunny, and of course, the fan favourite, the Cursed Bird.

The Summer collection features a Flamingo (Rhodotorula glutinis), a Prickly Pear Cactus (Pseudomonas aeruginosa) and a slice of Watermelon (Pseudomonas aeruginosa & Pseudomonas spp.).

Thank you for following my inquiry blog this semester, and good luck with your studies!

In the spirit of the up coming holidays, here is some festive agar art!

Here is an Escherichia coli (E. coli) snowflake on Tryptone Glucose Yeast Extract Agar with MUG Supplement. When digested, MUG produces a fluorescent molecule which glows under UV light!

Here is an infographic detailing UV positive and UV negative bacteria on TGEA w/ MUG Supplement.

Lastly here is some not-so-sweet candy canes using Serratia marcescens (Red) and unknown bacteria (White).

Thank you for following me along this inquiry journey!

Pseudomonas aeruginosa

Over the course of this inquiry, I have really enjoyed working with Pseudomonas aeruginosa, and it has become my favourite organism. Pseudomonas aeruginosa is an opportunistic pathogen, meaning that in most cases, it is harmless to the healthy individual. Cultures of Pseudomonas aeruginosa smell good. If you’ve ever stepped foot in a microbiology lab you’ll understand that good smells and bacteria are on the complete opposite sides of the spectrum, so working with a microorganism that smells good is a welcomed luxury. Pseudomonas aeruginosa grows fantastic green colonies that glow under ultraviolet light! Since Pseudomonas aeruginosa is my favourite microorganism, I figured it would be fitting to create some of my other favourite creatures, the dinosaurs!

For these dinos, I printed off a picture, taped it to the underside of the plate, and traced it using my inoculating loop with the help of a dissecting scope. I incubated them at 37^oC for 24hrs and they turned out better than I ever expected!

Today I’m going to shift focus to a organism that I am specialized in. This bacteria is called Legionella pneumophila, and it’s claim to fame occured in Philadelphia in 1976/77. Hundreds of goers from the Legionnaire Convention fell ill with symptoms of pneumonia, resulting in the deaths of 15% of those people who showed symptoms. The CDC was brought in to investigate and their extensive investigation came up empty. They couldn’t find presence of viral, fungal or bacterial agents in the hotel, and there were no toxins or heavy metals present in the autopsies. The CDC was stumped. Luckily the outbreak seemed to subside, and things returned to normal, but scientists and researchers were reluctant to give up on this mystery disease.

Joseph McDade, a microbiologist who specialized in intracellular bacteria, decided to use a culturing method for intracellular bacteria, a method that is not commonly used. He discovered bacteria growing which wasn’t present before. This bacteria was called Legionella pneumophila.

As a microbiologist, I am specialized in how to culture, identify, serotype and report out cases of Legionella pneumophila. I was a project leader and was responsible for testing Legionella all across the province. It was a fascinating experience and my knowledge in the scientific field grew exponentially. Working with Legionella was definitely not easy. It really allowed me to develop problem solving and critical thinking skills, made me become spatially and organizationally cognisant, and it really made me learn how to be patient.

Legionella cultured on specialized agar called BCYE in an low-oxygen environment presents itself as small, slow-growing colonies that have a slight blue undertone and an appearance of shattered glass.

Legionella species grow very slow, and they require specific conditions in order to grow. One instance of this, is Legionella requires the presence of the amino acid, cysteine, to grow. In the picture below, I had four cultures of suspected Legionella, and only one of the agar plates contained cysteine. The positive Legionella culture was unable to grow on the media without cysteine, leading to me to take further measures to determine whether or not it was truly Legionella.

The next step is to do serology to determine the species of Legionella. This is done through an antibody test. Some of the bacterial colony is scooped up and homogenized with saline, and it is then mixed with antibodies that target proteins on the outside of the bacteria. The antibodies will start to bind to Legionella, clumping them together and eventually precipitating it out of the saline. Here is an example of a positive serology test of Legionella pneumophila!

Next week I’ll return back to agar art!

I’m excited to showcase two agar art pieces in the spirit of Halloween!

Staphyl-o-lantern aureus:

This agar art was made on the orange coloured Vogel-Johnson Agar using Staphylococcus aureus. I had to carefully time this piece, as S. aureus ferments the sugar mannitol into an acidic byproduct. The acidity causes the pH indicator in the agar to turn yellow- meaning that shortly after i took this photo, the agar turned yellow. The agar also contains tellurite which is reduced by S. aureus into metallic tellurium, which causes the bacteria to turn black!

Now the scary part about this piece isn’t the mischievous grin or the spooky eyes, but the organism I used. In medicine, a strain of Staphylococcus aureus called “MRSA”is one of the emerging pathogens that have become resistant to an entire class of antibiotics and shows increased resistance to other classes of antibiotics. This poses a huge problem in hospitals, as healthcare professionals struggle to treat MRSA with their limited antimicrobial toolkit. Luckily for me, this is unlikely to be MRSA.

Pseudomona AHH-rugenosa

This agar art was made on Pseudomonas Isolation Agar using Pseudomonas aeruginosa. Glycerol is used as the primary carbohydrate source, which helps promote the production of a ghastly green pigment called phycocyanin. Antibiotics present in the agar prevent growth from other species outside of the Pseudomonas family.

Pseudomonas aeruginosa produces a metabolite that is fluorescent under ultraviolet light, causing it to glow a distinct aqua green. Pseudomonas is weird in a way where it gives off a pleasant fruity smell which I would describe as either pear or grape. I cannot confirm if it tastes any good, so maybe stick to Halloween candy this year!

Happy Halloween!

Working with living organisms is difficult because it must continually be cared for and renewed, or frozen in liquid nitrogen to preserve it for long periods of time. For ease of access, I keep an archived collection of bacterial colonies in a liquid nutrient-rich media and transfer them into new media on a weekly basis. Maintaining this can be tedious and time-consuming, but is necessary for bacterial survival, as they could potentially die off due to lack of nutrients and build-up of waste products if I don’t.

I transfer this bacterial suspension to nutrient agar plates as well, so I have “paint” to create art with as solid colonies is much easier than liquid suspension. This agar plate serves as my microbiological easel, a palette of colonies of different colours I can use to produce art.

Instead of subjecting everyone to another wall of text, I have created some graphics detailing the process of culturing and

So how does one start? Once you have your mature bacterial colonies and agar plates ready to use, it is time to get to work! The inoculating loop is a tool used in labs which are used to transfer bacteria between plates and media. In the context of art, the inoculating loop is the microbiologist’s paintbrush.

Due to the small size of the agar plates, I will be using a dissecting scope to help me make sure my edges are sharp and that I do not cross-contaminate bacterial species while I’m painting. Transferring bacteria onto an agar plate is an invisible process at first, as the colonies you paint on are virtually impossible to see and requires concentration to ensure you are not overlapping any areas or you may risk contamination.

Comparison of cross-contaminated versus isolated:

Complete agar art before incubation. Can you tell what it is?

Next week I will be working with Staphylococcus aureus and Pseudomonas aeruginosa to create some Halloween themed art pieces. I am super excited for next week when I can showcase some SPOOKY art!

In the spirit of a new NHL season starting, I created a few NHL team logos as my art pieces! I created 5 NHL team logos from different bacterial and yeast species, as well as different agar plates, and they all turned out in various degrees of success!

Before I showcase my art, I’d like to tie in the relationship between hockey and microbiology. There is a tool we use in microbiology called a cell spreader. It is commonly referred to as a “hockey stick” and it’s primary use is to evenly spread a volume of bacterial suspension across the agar plate. I provided an image of the cell spreader hockey stick in the header!

To

The first plate I created is my favourite team, the Vancouver Canucks. I’ve been a huge fan of the Canucks since as long as I can remember, and I get excited for the start of every season despite how much misery they bring me. I created the original Canucks on mFC agar using E. coli (Blue) and Enterobacter species (White).

On the second plate, I used Serratia marcescens (Red) and E. coli (White) on nutrient agar to make the New Jersey Devils.

For the third plate, I used Serratia marcescens (Red) and Micrococcus luteus (Yellow) on nutrient agar to make the Calgary Flames.

For the fourth plate, I used Klebsiella pneumoniae (White) on mFC agar to make the Tampa Bay Lightning.

On the final plate, I used Serratia marcescens (Red), Rhodotorula (Pink), and E. coli (White) on nutrient agar to make the Montreal Canadiens.

Let me know your favourite!

Next week, I’ll continue into the science of Agar Art and then finish of with a spooky surprise for Halloween!

Bacteria are living organisms that require specific nutrients and environments to grow. To facilitate this, in the lab we use a Jell-O-like substance called agar which is made from red seaweed. Agar is mixed with sugars, water, and nutrients to create a medium that contains the necessary metabolic needs to support microbial life. Molten agar is poured into petri dishes or tubes, and solidifies as it cools. To answer the question that all new microbiologists ask… no, it does not taste as good as Jell-O. Another thing I would like to address is that agar is commonly referred to as “agar plates” or “plates”, so I’ll be using these terms interchangeably.

The most common type of agar is called “nutrient agar”, and it contains a rich source of common nutrients that support a wide variety of bacteria. Some bacteria are picky however, and require more or less of certain nutrients, or special nutrients not found in nutrient agar in order to grow.

Two other classes of agar are called “selective agar” which often contains inhibitors that prevent growth of specific bacteria, and “differential agar” which uses dyes and indicators to differentiate the growth of specific bacteria. Often these two classes are combined into one, which we call selective-differential agar.

While selective-differential media produce vivid colours, the colours are a direct product of substrates in the agar. On these types of media, only a few colours can be produced.

Bacteria on nutrient agar however produce pigments naturally, but it is much more rare to see vivid colours. Most bacteria are white, cream or beige looking, but I have found red, pink, green, yellow, orange, gold, blue and purple! Unfortunately I wasn’t able to preserve all of the colours, as some really struggled to grow with subsequent subcultures.

If you are curious about agar, here is a link to a useful resource!

Next week I will showcase some Agar Art!