I would first like to introduce myself. My name is Brandon Tompkins and I am currently going into my senior year at Pace University (Pleasantville campus) as a biology major. My goal this summer is to continue learning techniques and critical skills that will benefit me in my future medical career. Additionally the goal is to investigate the remarkable symbiotic relationship between bacteria and the Hawaiian bobtail squid. I would like to thank Dr. Andrew Wier for allowing me to work under his guidance in his lab. I would also like to thank Pace University for allowing me the opportunity to grow as a student. I wish everyone doing summer research the best of luck!!

 

 

The Euprymna Scolopes better known, as the Hawaiian bobtail Squid is part of the Sepiolidae family. The Hawaiian bobtail squid is native to the central Pacific Ocean, where it is found in shallow coastal waters off the Hawaiian Islands, hence the name! Though its beauty is recognized for its ability to light up the coastal waters much like a “fire fly” does the summer night sky. The reasoning for it bioluminescent is much more intriguing. The Hawaiian bobtail squid has become the backbone for the definition of a symbiotic relationship. That is a close prolonged association between two or more different organisms of different species that may, but not necessarily, benefit each other. In our case we know for sure from prior research that the squid is benefiting.  The benefit is the squid being colonized by the bacteria Vibrio fisheri (V. fisheri). V. fisheri has been carefully studied for it’s  association with the Hawaiian bobtail squid allowing the squid the ability to cloak itself from predators by matching the light intensity of the moon. In return the squid feeds the bacteria. [1][2].  Though this symbiotic relationship has been well studied there are many other relationships that are occurring in the squid that haven’t been.  Dr. Wier and I are focusing on the bacteria that colonize the Accessory Nidamental Gland (ANG) shown in Figure 1.  The ANG is found in female Hawaiian bobtail squid and is responsible for the secretion of eggs. Prior to the squid being sexually mature the ANG is a white color but once mature the ANG changes to red. This change is because of the bacteria colonizing the ANG at the time.  Our over arching question is what competitive nature may be exhibited amongst bacteria when colonizing the ANG? Our goal is to reproduce these possible competitive behaviors in a lab setting that can help us better understand bacteria’s role in vying for a spot on the ANG.

Figure 1:

Figure 1:  Image provided by The Nyholm Lab at The University of Connecticut. Image shows both the light organ where V. fisheri is found, and the Accessory Nidamental Gland (ANG) is shown next, which houses the bacteria of our interest.

Currently in the lab I have been working with bacteria Dr. Wier has previously collected from the ANG. I have grown these samples on media to ensure their purity.  I am working on running the bacteria through PCR so the DNA can be amplified enough for a clean up kit and sent out for sequencing.  I also have been working on examining the bacteria growth and inhibition when plating alone and with other bacteria that have been inoculated from the ANG. Dr. Wier and I have even gone as far as researching any possible inhibition when growing our unknown bacteria with some known ones like Escherichia coli (E. coli), Micrococcus luteus (M. luteus), and Bacillus cereus (B. cereus).

 

Figure 2:

 

Figure 2: ANG RO7 shows one of the first inoculated samples streaked out. The next step was to separate these potentially different bacteria from each other. When inoculating these different pigmented bacteria from each other it was surprising to observe the orange-pigmented bacteria disability to grow as well alone, as the white- pigmented bacteria. Figure 3: ANG RO7 white inoculated alone and Figure 4: ANG R07 orange inoculated alone shows difference in growth over the same period of time. Though this potentially could be a good find it must be replicated many more times.

Figure 3:                                                                                                 Figure 4:

                                                         

Figure 5:

Figure 5: shows the first PCR ran this summer. It was a success. Though it was a success it does not guarantee pure samples. Currently I am working on running a PCR and Gel Electrophoresis on potentially pure samples so it can be used for DNA cleanup and then sent out for sequencing.

 

Figure 6:

Last but not least Figure 6: shows myself figuring it out. It’s amazing how one over arching question can lead to a bunch of smaller questions. It never ends. I wish all other summer grant research receipts best of luck and may science be on your side!

 

 

References

 

  1. 1.    Reid, A. & P. Jereb 2005. Family Sepiolidae. In: P. Jereb & C.F.E. Roper, eds. Cephalopods of the world. An annotated and illustrated catalogue of species known to date. Volume 1. Chambered nautiluses and sepioids (Nautilidae, Sepiidae, Sepiolidae, Sepiadariidae, Idiosepiidae and Spirulidae). FAO Species Catalogue for Fishery Purposes. No. 4, Vol. 1. Rome, FAO. pp. 153–203.
  2. 2.    Lemus, J.D. & M.J. McFall-Ngai 2000. Alterations in the protoeme of the Euprymna scolopes light organ in response to symbiotic Aliivibrio fischeri. Applied and Environmental Microbiology