BACTERIA MORPHOLOGY CHARACTERISTICS TO STUDY BACTERIAL DIVERSITY.

Purvi kakrani Dr. Harish kakrani Dr. Bhanu Kakrani
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Bacteria…?
Leptospira, causes serious disease in livestock
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Bacteria are often viewed as the cause of diseases in humans and animals. Some bacteria are useful, for example certain bacteria aids in digestion. Bacteria make up the base of the food web in many environments. Bacteria are of such immense importance because of their extreme flexibility, capacity for rapid growth and reproduction, and great age. They can be photosynthetic, using light, or chemosynthetic, using inorganic chemicals as the source of energy, but most are heterotrophic, absorbing nutrients from the environment.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Background Information
Prokaryotes

Prokaryotes represent two domains, bacteria and archaea. Archaea live in Earth’s extreme environments. Bacteria are the most abundant and diversified organisms on Earth.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Bacterial Structure

Biochemical processes that normally occur in a chloroplast or mitochondrian of eukaryotes will take place in the inner membrane of prokaryotes.

Bacterial DNA is circular and arrayed in a region of the cell known as the nucleotide .
Scattered within bacteria’s inner membrane are numerous small loops of DNA known as plasmids .

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Structure

Some bacteria have flagella with a different microtubule structure than the flagella of eukaryotes.. Ribosomes are the structures in cells where proteins are assembled. Bacterial ribosomes have different sized ribosomal subunits than do eukaryotes.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Bacteria Have One of Three Cellular Shapes

Rods (bacilli)

Coccoid-Shaped

Spirilla

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Reproduction

Prokaryotic cell division is binary fission.
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Single DNA molecule that first replicates. Attaches each copy to a different part of the cell membrane. Cell begins to pull apart. Following cytokinesis, there are then two cells of identical genetic composition.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Now…On to our experiment...

Purpose: Identify varieties of bacterial colonies and investigate bacterial species diversity, by isolating, culturing, and analyzing bacterial colonies, or species, that inhabit:
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Air Pond Water Raw Chicken Washed/Unwashed hands Keyboard Soil Sample

Hypothesis: Knowing that bacteria can thrive in almost anywhere on our planet, we reason that all of the environments tested will grow bacterial species. We further hypothesize that the thumb print of the washed hand with the anti-bacterial soap, should house less species than any others tested, because the anti-bacterial soap should kill off all bacteria.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Methods

A .


B.

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C.


Figure 1.

Streak Plate Method. (a) Streak the plate back and forth across top half of plate. (b) Rotate plate a quarter turn counter clockwise and streak top right quarter of plate. (c) Rotate plate a quarter turn counter clockwise and streak top right quarter of plate again. 4/13/2012

For chicken, soil, pond water, and keyboard samples, streak the plate using the streak plate method to isolate bacterial colonies. Leave agar plate open for air sample. For the unwashed hand gently press thumb against agar. Take washed hand and gently press thumb against agar. Wrap in Parafilm and incubate the cultures for about one week at 22 C. Observe and Interpret Data

Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Results: Soil
SIZE
4 mm 3 mm 5 mm

SHAPE
Irregular Irregular Filamentous

MARGIN Lobate Lobate Filamentous

SURFACE Wavy Wrinkled Wrinkled

COLOR Yellow/white Brown/yellow Green/white

1

2

3

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Results: Pond Water
#
1 2 3

Size Shape
2 mm round 2 mm round 1 mm round

Margin
smooth lobate Smooth

Surface Color
Smooth contoured Smooth grey beige clear

2. 1. 3.

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Results: Raw Chicken
# 1 2 3 4 Size 2mm 3mm 1mm 2mm Shape Irregular irregular round Irregular Margin lobate lobate Smooth wavy Surface contoured wrinkled Smooth contoured Color yellow/green clear/white Yellow/green brown

1. 2. 4. 3.

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Results: Air
# 1 2 3 4 5 6 7
1 1 7 6

Size 5 mm 5 mm 1 mm 3 mm 3mm 5 mm 8 mm

Shape Irregular Round Irregular Irregular Irregular round irregular

Margin Surface Color Smooth Smooth Yellow/orange smooth contoured Yellow/orange Wavy contoured Yellow/white Lobate wrinkled Yellow/brown Lobate smooth White Smooth Smooth White/yellow lobate contoured White/yellow

2

3 4 5 5

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Results: Washed Hand

1. 2.
Size # 1 4 mm 2 1 mm Shape Irregular filamentous Margin Surface Color yellow white

lobate Smooth filamentous smooth

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Results: Unwashed Hand
Size # 1 2 mm 2 2 mm 3 1 mm Shape Irregular round round Margin lobate Smooth Smooth Surface Color smooth smooth Smooth yellow yellow white

1 2 3

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Results: Keyboard
Size Shape Margin Surface Color yellow white # 1 4 mm Irregular 2 1 mm filamentous
# Size 1 2 mm 2 1 mm Shape Irregular round Margin lobate smooth Surface Color wrinkled yellow smooth greenish

lobate Smooth filamentous smooth

2.

1.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Control

An unopened agar nutrient plate, which ruled out agar contamination, had no bacteria species present.

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Species vs. Environments
Number of Bacterial Species on Agar Plate
8 6 # of Species 4 2 0 Series1 Air Keyb Unw Was Pond Chic Soil 7 2 2 3 3 4 3

Environments

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Conclusions/Observations
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The results supported our hypothesis since bacteria grew in all of our samples. The results did not support our hypothesis concerning the hand washed with anti-bacterial soap since it did not house less species than the other environments tested. We were surprised to learn that the air not only housed the most bacteria, but housed the most bacterial diversity of species as well.
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Further Investigations…

Further studies can be conducted by using TEM microscopy, SEM microscopy, and gram staining, to specifically identify what type of bacterial species were present in each environment. Research can also be conducted to figure out as to why the unwashed hand contained more bacteria than the washed hand. Further research can be done to determine if any of the bacteria found in our samples are harmful to humans.
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Questions to Ponder…
Do all bacteria grow at the same rate, and what factors in the environment contribute to determining their “carrying capacity?”  What research can be done to determine whether bacterial species and fungus compete with each other for nutrients and space in selected environments?

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References
Coccoid-shaped Bacterium (causes skin infections), Enterococcus faecium (SEM x33,370). This image is copyright Dennis Kunkel at www.davidkunkel.com, used with permission.  Morgan, I.G. and Brown Carter, M. E., Investigating Biology: A Laboratory Manual for Biology. California: Benjamin/Cummings Publishing Co., Inc. 1993.  Rod-Shaped Bacterium, hemorrhagic E. coli, strain 0157:H7 (division) (SEM x22,810). This image is copyright Dennis Kunkel at www.davidkunkel.com, used with permission.  Spirilla- shaped Bacterium (SEM x33,370). This image is copyright Dennis Kunkel at www.davidkunkel.com , used with permission.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Serratia marcescens

Gram stain Motility Habitat Pathogenicity:
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Negative. Motile. Occurs naturally in soil and water as well as the intestine. Associated with urinary and respiratory tract infections, endocarditis, wound infections, and eye infections.
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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

Gram-positive and gram-negative bacteria

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Difference Between Gram-Negative and Gram-Positive Bacteria

Gram-Negative Bacteria

Gram-Positive Bacteria

More complex cell wall.

Simple cell wall.

Thin peptidoglycan celll wall layer.

Thick peptidoglycan celll wall layer.

Outer lipopolysaccharide wall layer.

No outer lipopolysaccharide wall layer.

Retain safranin.

Retain crystal violet/iodine.

Appear pink/red.

Appear blue/purple.

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Antibiotic Sensitivity Test

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Purvi kakrani, Dr. Harish Kakrani & Dr. Bhanu Kakrani

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Antibiotic Sensitivity Test

P10 TE30

Ø10

E15

C30

Ø14

Ø20

K30 NA30 Ø14 Ø8

S10

NB30

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Hypothesis: Kanamycin is one of the most sensitive antibiotics because infections treated include respiratory tract, urinary tract, skin, soft- tissue and abdominal infections. Prediction: The size of the zone of inhibition is the largest. Results: The size of the zone of inhibition is the 2nd largest
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Antibiotic Resistance

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Some bacteria have developed resistance to antibiotics naturally. Bacteria can become resistant to drugs in a number of ways. - Mutation. - Exchange genes with other bacteria. - Resistant traits spread to future generations quickly because of rapid reproducing.
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Mechanism

Antibiotics kill or stop the growth of harmful bacteria.

Limitations
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Reason unknown why S. marcescens is sensitive to certain medications. Further research needed.

Future Work

Develop new drugs to confront bacteria resistance.
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