A Broad-spectrum mer Operon in a Multi-drug Resistant Strain of the Fish Pathogen, Aeromonas salmonicida.

Aeromonas salmonicida AS03, a potential fish pathogen, was isolated from Atlantic salmon, Salmo salar, in 2003. This strain was found to be resistant to ≥1000 mM HgCl2 and ≥32 mM phenylmercuric acetate as well as multiple antimicrobials. Mercury (Hg) and antibiotic resistance genes are often located on the same mobile genetic elements, so the genetic determinants of both resistances and the possibility of horizontal gene transfer were examined. Specific PCR primers were used to amplify and sequence distinctive regions of the mer operon. A. salmonicida AS03 was found to have a pDU1358-like broad-spectrum mer operon, containing merB as well as merA, merD, merP, merR and merT, most similar to Klebsiella pneumonaie plasmid pRMH760. To our knowledge, the mer operon has never before been documented in Aeromonas spp. PCR and gene sequencing were used to identify class 1 integron associated antibiotic resistance determinants and the Tet A tetracycline resistance gene. The transposase and resolvase genes of Tn1696 were identified through PCR and sequencing with Tn21 specific PCR primers. We provide phenotypic and genotypic evidence that the mer operon, the aforementioned antibiotic resistances, and the Tn1696 transposition module are located on a single plasmid or conjugative transposon that can be transferred to E. coli DH5α through conjugation in the presence of low level Hg and absence of any antibiotic selective pressure. Additionally, the presence of low-level Hg or chloramphenicol in the mating media was found to stimulate conjugation, significantly increasing the transfer frequency of conjugation above the transfer frequency measured with mating media lacking both antibiotics and Hg. This research demonstrates that mercury indirectly selects for the dissemination of the antibiotic resistance genes of A. salmonicida AS03.

A broad-spectrum mer operon in a multi-drug resistant strain of the fish pathogen, Aeromonas salmonicida

Aeromonas salmonicida AS03, a potential fish pathogen, was isolated from Atlantic salmon, Salmo salar, in 2003. This strain was found to be resistant to ≥1000 mM HgCl2 and ≥32 mM phenylmercuric acetate as well as multiple antimicrobials. Mercury (Hg) and antibiotic resistance genes are often located on the same mobile genetic elements, so the genetic determinants of both resistances and the possibility of horizontal gene transfer were examined. Specific PCR primers were used to amplify and sequence distinctive regions of the mer operon. A. salmonicida AS03 was found to have a pDU1358-like broad-spectrum mer operon, containing merB as well as merA, merD, merP, merR and merT, most similar to Klebsiella pneumonaie plasmid pRMH760. To our knowledge, the mer operon has never before been documented in Aeromonas spp. PCR and gene sequencing were used to identify class 1 integron associated antibiotic resistance determinants and the Tet A tetracycline resistance gene. The transposase and resolvase genes of Tn1696 were identified through PCR and sequencing with Tn21 specific PCR primers. We provide phenotypic and genotypic evidence that the mer operon, the aforementioned antibiotic resistances, and the Tn1696 transposition module are located on a single plasmid or conjugative transposon that can be transferred to E. coli DH5α through conjugation in the presence of low level Hg and absence of any antibiotic selective pressure. Additionally, the presence of low-level Hg or chloramphenicol in the mating media was found to stimulate conjugation, significantly increasing the transfer frequency of conjugation above the transfer frequency measured with mating media lacking both antibiotics and Hg. This research demonstrates that mercury indirectly selects for the dissemination of the antibiotic resistance genes of A. salmonicida AS03.

Sculptural abstraction of landscape

The purpose of my Senior Scholar project was to create a series of sculptures that are based on the interaction of natural forms within a selected landscape setting. I hope to convey a sense of how I view and interpret the landscape and to create works that stimulate a sense of wonder in the mind of the viewer. This fascination, perhaps even obsession, with the power of the landscape has kept me going throughout the year. As a source of ideas and imagery, the landscape can never be depleted. There will always be new combinations of landscape elements, different light conditions, and changing seasons to provide me with fresh ideas. Research for the project took me to many different places, be it my trip to Monhegan Island or driving to New York City to study the cityscape. I began the year working in steel and plaster, combining the two in works that explore the interaction between living tree roots and inanimate rocks. This led to a series entitled Landscape Recollections, comprised of welded steel forms housed in protective wooden boxes and lit from inside. After visiting New York City, architecture began to playa role in my work, as seen in the two Roadcut pieces and the Cathedral Woods series. Roadcut #1 and Roadcut #2 explore the relationship between a man-made road and the landscape that lies beneath and around it. The Cathedral Woods pieces incorporate architecture in a more abstract manner, using imagery derived from Gothic cathedrals to convey a sense of quiet peacefulness. The soaring verticality of Gothic architecture integrates with the tall tree forms in each piece, enabling me to intertwine these two elements into one another and create a harmonious relationship between architecture and nature. Throughout the year I have kept a sketchbook in which I draw from life, jot down ideas, and take notes on how the project is progressing. I have also completed several large charcoal drawings of my sculptures in which I explore different ways of viewing the steel forms. I am grateful to have had the opportunity to undertake this project, as it has been a very difficult but rewarding process of observing, interpreting, and manifesting the landscape according to my personal vision and experiences.