Biofilms A Layer of life with so much

Biofilms A Layer of life with so much potential J. Adam Frederick and Jackie Takacs Maryland Sea Grant Extension Program Maryland Governor’s Academy 2008

Biofilms are composed of bacteria that create layers of material typically composed of sugar (polysaccharides) and fat (lipids) that adhere to surfaces in both dry and wet climates. These substances produced by the bacteria are know as EPS or extracellular polymeric substances. Biofilms are an important link to many nutrient cycles in natural communities. These organisms form the basis for food webs that nourish larger organisms such as insect larvae which are consumed by fish that are in turn consumed by birds, amphibians, etc.

Where do Biofilms Exist? Biofilms are found almost everywhere on a wide variety of substrate in both terrestrial and aquatic ecosystems under a wide range of abiotic conditions. Non-living substrate Plants Organisms (externally & internally)

Where are your Biofilms? We are perfect factories for biofilm development, growth, and expansion. We are highly symbiotic with a variety of bacteria, not all beneficial but not all bad either. Information about human flora can be found at: http: //textbookofbacteriology. net/normalflora. html

Cellular relationships Symbiosis A relationship that has developed between 2 systems (organisms, cells, etc) over time. Mutualism both organisms benefit from the relationship (lichen) Commensalism one organism benefits and the other is unaffected? Parasitism one organism is harmed by the relationship (tick)

Mechanisms of biofilm transfer External- symbionts introduced from the environment (bobtail squid) Horizontal- symbionts introduced by parents (E. coli in the gut) Vertical- symbionts carried by oocytes to next generation (some clams)

The good The human gut is host to many billions of microbes. Each gram (about a thimble -full) from the large intestine contains up to ten trillion (10, 000, 000) microbes! Microbes in the intestines perform many essential tasks. For example, microbes make several vitamins in the intestines. One of these is vitamin K, a vitamin that is otherwise lacking from human diets. Lactobacillus acidophillus This is a normal, helpful inhabitant of intestines. The billions of Lactobacilli in your intestines crowd out harmful microbes.

The good & bad E. coli This is the most well-studied organism. It is a normal resident of your lower intestine and provides you with vitamin K and some of the B vitamins. Bacteroides This is one of the most abundant microbes in the intestine, and is found in higher numbers in people who eat meat than a vegetarian diet. These species cause of a number of infections throughout the human body.

The ugly Clostridium botulinum “botox” Clostridium tetani

Biofilm formation in the Marine Environment The Sequence of Biofouling in the Sea R. E. Baier, Ph. D. IUCB, University of Buffalo, NY

Part I Spontaneous deposition of a primer coat of “humic acid of the sea”.

Part II Selective binding to the “primer” coat by rod shaped bacteria.

Part III Long-tailed microorganisms dominate the surface at 3 days and produce slimes as shown by the gray areas.

Part IV The extrusion of slime patches continues and the accumulation of metals may become evident.

Part V Long membranous tails are extended to accumulate nutrients. Other bacteria continue to attach to the layer.

Part VI Attachment of algae spores, diatoms, and larval forms of sessile inverts to filamentous appendages and slime.

Part VII After 1 week of exposure, entrapped and attached organisms begin the process of mineralization and binding.

A Focus on Marine biofilms

Molecular identification and localization of a filamentous symbiotic bacteria associated with the hydrothermal vent annelid, Alvinella pompejana The fuzzy white coat is not the typical setae of annelids but, symbiotic bacteria. S. C. Cary, M. T. Cottrell, J. L. Stein, F. Camacho and D. Desbruyeres. 1997, Appl. Environ. Microbiol. 63: 1124 -1130.

A. pompejana lives in an extreme temperature gradient unlike any other organism on earth. top of tube 2°C 80°C base of the tube

Charles R. Fisher PSU, Deep Sea Laboratory The bacterial symbionts of the tube worm make food for the worm through chemosynthesis of hydrogen sulfides.

The symbiosis exhibited by A. pompejana and the bacterium is still under investigation. The study of the eurythermal enzymes holds great promise for industry.

Charles R. Fischer Deep Sea Laboratory Penn State University Mussels located at a methane seep provide new insights into symbiosis at the deep sea level.

Charles R. Fisher PSU, Deep Sea Laboratory The gill tissue of the mussel is a prime location for the growth of bacterial symbionts and chemosynthesis.

Charles R. Fisher PSU, Deep Sea Laboratory SEM of bacterial symbionts from mussel gill tissue. Mussels have an in-house supply of food by consuming the bacteria and the products of chemosynthetic activity.

“Ice worms” that live on the methane hydrate mounds are under investigation. Charles R. Fisher PSU, Deep Sea Laboratory

Hawaiian reef squid or bobtailed squid, Euprymna scolopes Vibrio fischeri enjoys a “mutually beneficial” relationship with Euprymna. The reef squid makes a new bioluminescent culture each day and expels the old one to maintain a creative night-time camouflage.

Understanding how the Euprymna/Vibrio symbiosis works will provide scientists with information about how bacteria and other microbes -whether harmless or pathogenic are able to recognize and colonize specific tissues in its host.

The relationship between zooplankton and cholera bacteria has presented a problem for drinking water supplies in third world countries, especially during El Nino years causing enhanced disease outbreaks.

biofilm Research @ usm Dr. Ron Weiner (UM College Park) has been studying biofilm forming bacteria for years and described a species H. adherans that is responsible for most initial biofilm formation especially in the marine environment. In addition, he has successfully identified bacteria essential for the breakdown of cellulose and chitin. For more information go to; www. hyphomonas. com Hyphomonas adherans

Question: If H. adherans is cultured on slides or discs that are placed into an aquatic ecosystem, will that have an impact on the rate of biofilm development and diversity of organisms? American Type Culture Collection (www. atcc. org) has H. adherans for sale- the same species identified by Dr. Ron Weiner. Marine agar 2216 or marine broth 2216 Temperature: 26. 0°C.

biofilm Inspiration Biofilms and Biodiversity http: //www. mdsg. umd. edu/programs/education/interactive_lessons/biofilm/index. htm Biofilms and Biodiversity on the web is an interactive lesson on the use of biofilm communities as a learning tool for teachers and students.

biofilm Inspiration Winogradsky columns The Russian microbiologist Sergei Winogradsky was one of the first to isolate soil microorganisms responsible for the conversion of N compounds in the soil, obtaining pure cultures of bacteria capable of the conversion of ammonia to nitrate. Winogradsky also studied the consumption of hydrogen sulfide gas by sulfur-oxidizing bacteria directly in their natural habitat. He introduced the concept of autotrophy in 1887. The resulting modern Winogradsky columns make excellent projects for studying anaerobic microbial activity in soil and the concept of autotrophy in the microbial world. http: //helios. bto. ed. ac. uk/bto/microbes/winograd. htm http: //ecosystems. mbl. edu/SES/Microbial. Methods/Winogradsky/default. htm http: //www. woodrow. org/teachers/esi/1999/princeton/projects/microbe/win_col. html

biofilm Inspiration Kombucha Tea Kombucha tea is produced by a mixed culture of bacteria and yeast and has been claimed by many a popular tea for detoxification. Culturing kombucha is an easy way to test bacterial activity under a variety of abiotic conditions and nutrient supplement without an incubator or pathogenic concern. There is some good culture information at http: //helios. bto. ed. ac. uk/bto/microbes/biofilm. htm#Top http: //www. kombu. de/anleit-e. htm http: //www. kombucha. org/

Digital biofilms Capturing biofilms through digital microscopy can be done in a high quality affordable manner that will also give teachers an additional instructional tool for a life-time with almost limitless potential. • Martin Microscope- adapter for connecting a video camera to the microscope (MM-99). Cost: $300 • Digital Video (DV) Camera- records to tape or hard drive and captures images on a memory card (e. g. Sony HDR-HC 5 or HDR-HC 7). Cost: $600 and up.