Small World Initiative Jason Heintskill

Kayla Ramnarine

Sarah DeMello

Caroline Lacagnina

BSC1005L: Life Science Lab

4/15/18

Small World Initiative

SWI combines technology and science to further our studies and improve the

healthcare field.

● SWI was developed by Dr. Jo Handelsman at Yale University in 2012

● Discovery-based biology course

● Students around the world contribute to creating new antibiotics.

What are microbes?

● Microbes are the source of many diseases, but also provide many benefits.

● Microbes are in and on our bodies and play a huge role in our overall health.

● Helps recycle nutrients

● Bioremediation

● Research tools

● Industrial applications

● ANTIBIOTICS!

Antibiotic Crisis ● No new classes of antibiotics have been discovered

since the 1980s.

● Over 2 million drug resistant bacterial illnesses

exist.

● $35 billion economic losses per year.

ESKAPE pathogens are six organisms that cause

devastating illnesses and most of the antibiotic resistant

infections.

● Chose 2 ESKAPE safe relatives (1 gram- 1 gram +)

Sarah Kayla Caroline Jason

Gram + S.epi S.epi S.epi S.epi

Gram - A.bay E.coli A.bay P.put

● Week later, checked for zones of inhibition

Sarah Kayla Caroline Jason

Zone yes/no Yes No No No

Testing Safe Relatives

Finding our Zone of Inhibition

● If we had at least one zone of inhibition, we streaked for singles

- If we had no zone of inhibition, we switched to a TA plate

● We received our PCR reactions to load in to

the gel

● Cut up the agar covered in bacteria

● Put dry ice in then pumped 10 ml of water and

15 ml ethyl acetate

Sim Tube

● Take from the same colony stab a needle

into the sim tube.

Sarah Kayla Caroline Jason

Motility Yes Yes No No

Catalase Production ● Pick one colony from your master streak

plate

● Add one drop of 3% H2O2 to the colony

smear

● See if our bacteria produced Catalase

Sarah Kayla Caroline Jason

Catalase

Production

Yes Yes Yes No

● Flood smear with Crystal Violet and Safranin

Gram Staining

Results Purple: Gram Positive Pink: Gram Negative

Jason ✓

Kayla ✓

Caroline ✓

Sarah ✓

Organic Extraction

Picture from : Viltes

powerpoint

Observing MacConkey Plates and Sim Tube

● If the growth on MacConkey agar was red = Lactose fermentation

● Black/Brown color on sim tube = Hydrogen sulfide production

● If the reagent turns cherry red = Metabolizing tryptophan.

Sarah Kayla Caroline Jason

Lactose

Fermentation Yes Yes Yes Yes

Hydrogen Sulfide

Production No No Yes No

Metabolizing

Tryptophan No No No No

Table 1:

Table 3: Table 2:

Data

Student Own Soil GPS

Coordinates

CFU/g Single Colonies

Sarah Yes Latitude:

26.3685

Longitude

:-80.1030

1.85E+07 Yes

Kayla No 26.3988889, -

80.20111111111

112

3.20E+06 Yes

Jason Yes Latitude:26.3680

Longitude

:-80.1040

1.92E+07 Yes

Caroline No 26.405277,-

80.217896

1.52E+07 Yes

Antibiotic Activity Results

Student Medium Temperature Light or Dark Antibiotic

Activity Results

Kayla BHI 30 Dark Bacillus subtilis

Jason TSA 28 Dark Staphylococcus

epidermidis

Sarah BHI 28 Dark None

Caroline BHI 28 Dark Bacillus subtilis

DNA Sequence Results

Jason Caroline Kayla Sarah

PCR Product

& Sequence

No, 124-TMJK-SWI27FHT

Yes,

22-CRL-SWI27FHT

Yes,

8-KNR-SWI27FHT

Yes,

Organism with

Closest Match to

DNA Sequence

Pseudomonas

sp. strain SB904

16S ribosomal

RNA gene,

partial sequence

Bacillus

Aryabhattai

strain WBY-6

16S ribosomal

RNA gene,

partial sequence

Bacillus cereus strain

AIMST 1.Cit.6 16S

ribosomal RNA gene,

partial sequence

Bacillus

aryabhattai 16S

ribosomal RNA

gene, partial

sequence

Percent

Identity

99% 98% 99% 98%

Bacteria’s Future Plans...

● What we could do in the future with our bacteria is now that we know they are

resistant to ESKAPE pathogens, the next thing to test is if they are resistant

to real pathogens.

● Test same soil with different conditions to see results.