Biofilm Research at COMB PowerPoint


Feb 21, 2013 (4 years and 5 months ago)


Exploring the Effects of

Antimicrobial Material on the Process
of Biofouling in an Aquatic Environment

Ms. Kathleen Tunney

Burleigh Manor Middle School, Howard County, MD

UMCES~Sea Grant Summer Research Fellow 2002

Mr. J. Adam Frederick

MD Sea Grant Extension/ COMB

Baltimore, MD

I worked with J. Adam Frederick of the MD Sea Grant Extension
Program and Frank Vavra of Maxxam Industries, Baltimore, MD, as
part of the Summer Research Fellowship at the Center of Marine
Biotechnology (COMB). The research project concerned the
effectiveness of an industrial anti
microbial material on the
prevention of biofouling.

Research Project

Classroom Project

One of my tasks during the summer research fellowship was to
develop an application for the classroom based upon my research
project. The activity I developed is entitled,

Culturing Biofilms and Exploring Biodiversity in Classroom Aquaria

Research Project

Mr. J. Adam Frederick

Ms. Kathleen Tunney

Exploring the Effects of

Antimicrobial Material on the Process
of Biofouling in an Aquatic Environment

Adapted from an educational outreach project by, J. Adam Frederick, D. Jacobs,
and W.R. Jones, Journal of Industrial Microbiology and Biotechnology (2000) 24,

I worked with J. Adam Frederick at the Center of Marine
Biotechnology as part of the Summer Research Fellowship in

Mr. Frederick’s work in the Inner Harbor in Baltimore,
MD, concerns the development of interactive educational tools
on the web related to biofouling and biodiversity.

COMB is situated on Pier 6 of the
Baltimore Inner Harbor and
provides easy access to the water
for the study of biofilms and the
process of biofouling.

A biofilm is a “coating or covering on the surface of a living or
nonliving substrate. It is composed of organisms like bacteria,
protozoa, algae, and invertebrate animals.” (Maryland Sea
Grant, Biofilms and Biodiversity lesson,

Biodiversity is the number of different species of organisms in
a particular environment and can change over time as
conditions change.

The research project during the summer of 2002 involved the
study of biofilms occurring on a variety of materials
submerged in Baltimore’s Inner Harbor. Biofilm development
and biodiversity on these materials were studied for two
consecutive 21
day periods: June 27 through July 17 and July
18 through August 7, 2002. Water quality tests and weather
conditions were monitored throughout each period.

Experimental Set

Test materials were in the form of 10cm diameter discs. The
materials were provided by Maxxam Industries in Baltimore, MD,
and were manufactured with a patented EPA approved anti
material. The material was either coated on the 10 cm discs or
embedded in the discs.

Test materials included control material (no anti
compound), 2% and 4% concentrations of anti
material applied as coatings on discs, and 2% and 4% embedded
in the discs.

Six discs of identical test material were stacked on a PVC post
or rack including controls. Four PVC racks of various test
material were hung vertically from a PVC pipe.

Four of these racks were suspended from Pier 6 in the
brackish water of Baltimore’s Inner Harbor.

Control and Experimental Samples

Collection of Sample Test Material

Racks were removed from the water every Monday and Wednesday
over a 21
day period. Two discs of each type (control and
experimental) were removed from the racks during each sampling
day and taken to the COMB lab for analysis. Collected discs were
viewed in the lab for biofilm development and biodiversity with
stereomicroscopes and compound microscopes.

Racks were returned to the water after discs were removed for
sampling. Water quality and weather conditions were recorded each

Analysis of Sample Test Material

Discs were observed in the lab for types of species and
numbers of individuals. Biofilms from each disc were
scraped with a razor blade and weighed for wet biomass
and then dried to measure dry biomass for comparison.

Biodiversity was calculated using the Maryland Sea Grant’s
online worksheet for calculating biodiversity after random
sampling procedures were followed for enumerating species
on each disc.

Go to the following for more reading:

Types of Organisms Identified

Whip mud worms







Clutches of fish eggs



Colonial ciliates

Stentor and other ciliate protists



Photos by Kathleen Tunney with Olympus America 35 mm microscope system

In addition to various lab skills developed over the course of the
week program, the opportunity existed to photograph
microscopic organisms while viewing them on the microscope.

This version of 35mm photography was not as easy as it sounds and
a greater appreciation for this area of photography occurred as a
result. Only one out of ever fifty so photographs taken represented
the organisms well. Digital photographs were taken in addition to
35mm pictures.

Microscopy & Photography




Representative Data Set

Dry Biomass of Biofouling Samples from Second Three Week Period.

Representative Data Set

Dry Biomass of Biofouling Samples from Second Three Week Period

The representative data set on the previous slide illustrates the
means of a large amount of raw data. Each data point on the graph
for a particular sample is composed of a larger number of smaller
samples which were analyzed and reduced to the mean value. The
data illustrates that no particular sample had a greater amount of
biofouling than another and that all samples followed a similar
pattern of development related to total biomass. Additionally, the
total amount of biomass was relatively low and resembled samples
from the acrylic control discs.

Acrylic controls are known to be highly “attractive” to biofilm
forming microorganisms and are used as a standard of comparison

Research Project Conclusions

Overall, much more work needs to be done in the field of biofouling
prevention since there are few options other than toxic marine
coatings available for the shipping industry, research vessels and
recreational boating. However, proper licensing is required to use
marine coatings.

The work we performed this summer also teaches that there are so
many environmental variables that relate to biofouling that the
process is very difficult to prevent without using toxics.


Ms. Kathleen Tunney, Burleigh Manor Middle School

Dr. Dan Jacobs, Maryland Sea Grant

Mr. Frank Vavra, Maxxam Industries, Baltimore, MD

Mr. J. Adam Frederick, Maryland Sea Grant


American Society for Microbiology

Microbial Slime,

Biofilms and Biodiversity web lesson, Maryland Sea Grant,

Frederick, J. Adam, D. Jacobs, and W.R. Jones,
Biofilms and Biodiversity:
an interactive exploration of aquatic microbial biotechnology and ecology
Journal of Industrial Microbiology and Biotechnology (2000) 24, 334

Wood, Timothy S.,

Aquarium Culture of Freshwater Invertebrates,
American Biology Teacher

vol. 58, January 1996.