Unknowns Project 9

25 November 2012

On this day, I finally identified my last unknown. It came from the shower plates I have been isolating. The isolation on the water agar plate looked microscopically like Bipolaris sp., but not macroscopically.  On the water agar, growth was very dark and sparse (Fig. 1). Bipolaris sp. is supposed to be thick and cottony macroscopically. When observed under the microscope, this fungus looks very much like Bipolaris, which has brown conidiophores and conidia, and conidia that have many cells (Figs. 2-3). 

Fig. 1
Shower mold isolated on water agar

Fig. 2
Conidia of putative Bipolaris

Fig. 3
Conidiophore of putative Bipolaris

When viewing this fungus on PDA, however, the macroscopic characteristics are completely different (Fig. 4) while the microstructure remain the same (Figs. 5-7). To me, this looks very much like the pictures and drawings I have seen of Bipolaris, so I am sticking with that identification.

Fig. 4
Putative Bipolaris grown on PDA

Fig. 5
Bipolaris sp. conidia

Fig. 6
Bipolaris sp. conidiophores

Fig. 7
Bipolaris sp. conidiophores

The last thing I did on this day was make another round of isolations for all of my unknowns just to make sure that I have them in pure culture to turn in on Wednesday. All isolations were made on 1/2 strength PDA.

All for now,

C

Unknowns Project 8

21 November 2012

For this day, I worked only on making pure cultures of my Curvularia and Pestalotia sp. cultures. I cultured from both my original plate (Fig. 1), and from isolations I had already made (Fig. 2). All isolations were made on 1/2 strength PDA. 

Fig. 1
Original plate

Fig. 2
Previous isolation with the left side being Pestalotia and the right side being Curvularia sp.

 All for now,

C

Unknowns Project 7

3 November 2012

Introduction

On this day, I observed many of my cultures macroscopically and microscopically, and was able to identify two genera. 

Observations

I first examined my wasp nest plate (Fig. 1), which contained many different colonies (Fig. 2). However, when observing these colonies microscopically, most of them appeared to be of the Aspergillus genus (Figs. 3, 4).

Fig. 1
Wasp nest plate 

Fig. 2
A variety of different colonies on the wasp nest plate

Fig. 3
Taken from wasp nest colony, appears to be Aspergillus

Fig. 4
Another sample taken from wasp nest colony, appears to be Aspergillus

One seafoam green colony, however, did appear to be different from the others microscopically (Figs. 5-9). 

Fig. 5
Conidia from seafoam green colony on wasp nest plate

Fig. 6
Conidiophore from seafoam green colony on wasp nest plate

Fig. 7
Conidiophore from seafoam green colony on wasp nest plate

Fig. 8
Conidia from seafoam green colony on wasp nest plate

Fig. 9
Yet another strange-looking conidia from seafoam green colony on wasp nest plate

 I next observed the cricket isolation plate (Fig. 10). I'm not sure about the conidiophores in Figs. 11-13, but I'm pretty sure that Figs. 14-16 show a common Zygomycete and perhaps Aspergillus.

Fig. 10
Cricket isolation plate

Fig. 11
Conidiophores from cricket isolation

Fig. 12
Conidiophores from cricket isolation

Fig. 13
Conidiophores from cricket isolation

Fig. 14
Putative Zygomycete

Fig. 15
Aspergillus sp?

Fig. 16
Aspergillus sp?

I next observed the shower mold again. This time, I was only looking at the plate which contained the agar block transferred from the original plate. When observing this under the microscope, it was clear that chlamydospores were present (Figs. 17-19). However, I was not able to identify the genus at this time. 

Fig. 17
Chlamydospores from shower plate

Fig. 18
Chlamydospores from shower plate

Fig. 19
Chlamydospores from shower plate

 Aaaannnnddd finally.....
At the very end of the day, I was finally able to catch a break by identifying two very easily recognizable genera (thank gods!).

First, I took the plate containing the large black fungus I scraped from a tree in Longview. I had not looked at this plate since I made it, so I was curious to see what I would find. It turns out that both Curvularia (Fig. 20) and Pestalotia sp. (Fig. 21) were present on the plate on separate sides. I apologize for the extremely poor quality of the photographs; my camera died right before I identified these, so I had to use my iPhone (which is very hard to use for this purpose).

Fig. 20
Curvularia sp.

Fig. 21
Pestalotia sp.

 The very last thing I did on this day was plate fungus from a piece of decaying wood that had green mycelia growing inside (Fig. 22). It appeared as though ants were farming this fungus. I also plated fungus from a watermelon leaf (Fig. 23). Both were plated onto 1/2 strength PDA.

Fig. 22
Decaying wood with green mycelia inside

Fig. 23
Watermelon leaf with white fungus on the surface

Conclusion
I am very happy to have 2/3 unknowns identified. Hopefully I can get the last one soon!

All for now,

C

Unknowns Project 6

31 October 2012

On this day, I observed many of my cultures and tried to identify some using Barnett and Hunter (2010). First I observed my culture from the wasp nest, which had several different colonies growing (Fig. 1).

Fig. 1
Wasp nest plate

 Next, I observed the plate taken from my moldy crickets (Fig. 2).

Fig. 2
Colonies isolated from moldy cricket

I then observed my plates isolated from the shower mold (Figs. 3-5). Fig 3 is the original plate, while Figs. 4 and 5 were isolated on water agar from the first plate. One plate was inoculated via standard sterile technique with a probe (Fig. 4), while the other was inoculated by sterilely cutting a chunk of agar from the original plate and placing it in the middle of the new water agar plate (Fig. 5). Both plates exhibit very sparse growth. 

Fig. 3
Original shower mold plate

Fig. 4
Fungi isolated from original shower mold plate using standard inoculation technique

Fig. 5
Very sparse fungal growth on plate isolated from original
shower mold plate via transfer of agar block

 Lastly, I observed the cricket mold under the compound scope. I did not have time to look at the other cultures under the scope. There were a few different types of fungi on this plate, from what I could tell. From a tape mount, I could clearly see that there was a mass of hyphae (Fig. 6). Looking more closely at some of the conidiophores, I could tell that the conidia had an unusual shape that was bulbous at the base and more narrow at the tip (Fig. 7). The other structures I saw looked like they were Aspergillus sp. (Figs. 8, 9).

Fig. 6
Mass of hyphae at 10x magnification (tape mount).

Fig. 7
One lone conidiophore

Fig. 8
Possibly Aspergillus sp.

Fig. 9
Possibly Aspergillus sp. 

Conclusions

Looks like I am still getting a lot of very common fungi. Maybe some of my other cultures will prove to be interesting.

All for now,

C

Unknowns Project 5

28 October 2012

On this day, I basically checked all of the cultures I had so far and looked at them under the microscope. The first culture I looked at came from some dark green colonies on the pig isolations (Fig. 1). These were not the same colonies as shown in the previous blog posts. Under microscopic examination, and with the help of Barnett and Hunter (2010), I believe this is some type of Zygomycete, most likely Mucor or Rhizopus (Figs. 2, 3). Boo!

Fig. 1
One of the dark green colonies from a pig isolation.

Fig. 2
Microscopic examination of the conidiophore of a dark green fungus

Fig. 3
Conidia of dark green fungus, most likely a Zygomycete

Next, I observed my plate taken from the shower (Fig. 4). I also observed this fungus under the microscope, but was unable to make an ID (Figs. 5-7). This particular fungus exhibited clusters of pill-shaped spores that exhibited at most 4 divisions.

Fig. 4
Plate taken from shower

Fig. 5
Conidiophores of shower fungus

Fig. 6
Condiophores and spores of shower fungus

Fig. 7
Conidiophores and spores of shower fungus

 The next culture I observed was the light green fungus isolated from the pig plates (Fig. 8). The fungus grew very well on the 1/2 strength PDA. At first glance, I thought this was another Zygomycete (Fig. 9) , but then I noticed that there were septa in the hyphae (Fig. 10). When viewing the entire plate under the compound microscope (Fig. 11), the shape of the structure become more clear (Fig. 12). By utilizing a tape mount, I was also able to see the structures better (Fig. 13). From these observations, I believe that this is an Aspergillus species (which I also cannot use).

Fig. 8
Light green fungus isolated from pigs

Fig. 9
Conidiophore at 40x magnification

Fig. 10
Hypha showing septa 

Fig. 11
Observation of entire plate under the compound microscope

Fig. 12
Observation of entire plate under the compound microscope

Fig. 13
Utilization of a tape mount in order to better observe structures

 After observing all of the fungi that I had cultured, I next attempted to inoculate some 1/2 strength PDA with fungi from a dead cricket (Fig. 14) and an abandoned wasp nest (Fig. 15). The fungus from the cricket was transferred to the PDA plate via sterile technique with an inoculation loop. The wasp nest fungus, on the other hand, was gently touched to a plate (just to see if anything would grow). Before these two samples were brought into the lab, the were kept in separate, sealed brown paper bags (just FYI).

Fig. 14
Moldy cricket found in a ditch

Fig. 15
Abandoned wasp nest

 Conclusion
Looks like I need to spend an ample amount of time trying to figure out all of my samples!

All for now,

C