The Journey

Zachary Bartow, Noah Quick, Alli Sanzari, Shereeka Brutis BSC 1005L-040 Life Science Lab April 18, 2018

http://www.smallworldinitiative.org Small World Initiative and Soil Microbes ● Find potential in soil for antibiotic crisis ● at an alarming rate becoming antibiotic resistant due overuse / resistant strains ● 14 countries, 38 states and Puerto Rico ● Mission : engage the youth in science and combat the antibiotic crisis

● Soil microbes - study of organisms in soil, their functions and http://birdcage3.blogspot.com/2013/11/a- how they affect soil properties misunderstood-form-of-art-agar-art.html ○ Ex : bacteria, fungi, algae, protozoa

https://www.ien.com/supply-chain/news/20988510/us-craft-beer-industry-is-exploding http://www.lucyswhey.com/cateringplatters3/deluxe-cheese-platter http://www.cwwiebemedical Antibiotic Crisis and ESKAPE .ca/-truly-allergic/

● Antibiotics - chemical that kills bacteria, inhibits the growth of microorganism ● Bacteria is becoming resistant to antibiotics due to overuse ● Pathogens are disease causing microorganism such as bacteria, virus, fungi, and protozoa ● ESKAPE pathogens - organism that are considered a major threat, make up the the majority of antibiotic resistant infections ○ ESKAPE pathogens ■ , , , , , and species ○ Safe relatives used in lab ■ Staphylococcus epidermidis, Escherichia coli, Erwinia carotovora, Acinetobacter baylyi, subtilis, Enterobacter aerogenes, and Pseudomonas putida Bacteria Basics

media Temp. Light or Isolate Gram Dark # stain (+/-)

Zach BHI 25 Light AEA26 NA

Noah BHI 25 Dark ADT6 +

Alli LB 30 Dark MEE26 -

Shereeka LB 30 Dark LRB9 + Results

Student Own GPS CFU/G Gram Single Catalase Tryptophan Motility Lactose Hydrogen soil coordinates + or Colonies Production Metabolism (tail) Ferment Sulfide (bubble) ation Gram Production -

Zach No 26.3832, - 4.9x10^7 NA Yes Yes No Yes Yes No 80.11464

Noah No 26 28’11’’N, 1.14x10^7 Yes Yes No Yes Yes No 80 8’56”W +

Alli No 26.3832, - 3.24x10^7 Yes Yes No Yes Yes No 80.11464 -

Sher No 26.370753, - 2.0x10^7 Yes Yes No Yes Yes No 80.101544 eeka + Macconkey Agar

MacConkey growth Ferment lactose Match Gram Staining results?

Zach yes yes No

Noah yes yes Yes

Alli yes yes Yes

Shereeka yes yes Yes Antibiotic Activity Results

Kills Resistant to

Zach P. put Gram, Tri, Rif, Tet, P/S

Noah B. sub, Gram, Tri, Rif, Tet, P/S P. put,

Alli B. sub, Gram, Tri, Rif, tet, P/S P. put

Shereeka N/A Gram, Tri, Rif, Tet, P/S DNA Sequence Results PCR Product and Organism with closest Percent Identity Sequence match to DNA Sequence

Zach EEE-SWI27FHT Bacillus megaterium strain 99% Edgemont-009 16S ribosomal RNA gene, partial sequence

Noah 10-ZBS-SWI27FHT Bacillus megaterium strain 98% Edgemont-009 16S ribosomal RNA gene, partial sequence

Alli 37-UBB-SWI27FHT Bacillus thuringiensis strain Bt 99% 7 16S ribosomal RNA gene, partial sequence

Shereeka MUH26S-P17- Pseudomonas sp. PONIH3 99% SWI27FHT chromosome, complete genome

Conclusion Future Direction ●More interested in bacteria ● Bacteria is everywhere experiments; looking to keep up with ● Can grow from the smallest thing Yale bacteria studies ○ Streaking for single colonies ●Testing the ESKAPE pathogens in a secure lab

Discussion

● Tested more single colonies (if any) ● Better original soil samples

https://www.123rf.com/photo_21699425_scientist-or-professor-looking-around- a-blank-sign-with-advice.html