The microorganisms in the biosphere are limited by nutrient availability, environmental conditions, energy, and carbon sources. Despite these limitations their role within a functional microbial community are perpetuated and distributed globally. In this study, we constructed anaerobic microcosms, enrichments, and purifications using serial dilutions from different environments to test the diversity and distribution of microorganisms that were able to use collagen as sole source of energy and carbon. The diverseenvironments included in this study are municipal waste water, rumen fluid content, UPRM cafeteria oil trap, pig food, sediments from the bottom of a water reservoir, sediments from an oxidation lagoon in a coffee plantation, sediments from a puddle with decaying grass, and sediments from a rice paddy. The collagen was added as a solid sponge and incubated at 37°C; its degradation was determined by the solubilization of the sponge compared to the control while the enrichment process was determined microscopically. Our results show that all environments except one (rice paddy) contain anaerobic microorganisms that liquefy and degrade collagen. Each environment responded to the stimulus at different time intervals with lag phases that ranged from days to weeks. Furthermore, microscopic observations of positive tubes from the different enrichments showed the same microbial morphotype present in all of the serial dilutions with increase abundance in the more diluted samples suggesting that an enrichment process occurred. In conclusion, it appears that the same organism based on microscopic morphological evidence is responsible for the degradation of collagen in the different environments tested.

Diversity and distribution of anaerobic collagen de graders in Tropical Environments.Luis A. Piñero García* and Luis A. Ríos Hernández

University of Puerto Rico at Mayagüez, Biology Depa rtment

In our research we want to test the hypothesis that “everything is everywhere, the environment selects” (M. Beijerinck). This implies that every environment has the potential to harvest microorganisms that will respond to a particular stimuli. Could we enrich different environments and obtain the same microorganism (species) or we will find the same functionality but different microorganisms? How do the environments selects for anaerobic collagen degraders? It is essential to know if there is any difference between the morphotype in those anaerobic collagen enrichments as function of the microbial community composition responsible of the degradation. In this study we made enrichments and utilize the serial dilution technique to enrich and purify the anaerobic collagen degraders from a diverse set of natural environments. In addition, the tubes that showed liquefaction of the collagen were analyzedmicroscopically to describe and compare the Prokaryotes that areinvolved in the degradation process.

H0: All environments contain the same anaerobic microorganisms that liquefy and degrade collagen.

H1: All environments contain different anaerobic microorganisms that liquefy and degrade collagen.

Lowry, OH., Rosebrough, NJ, Farr AJ, and RJ Randall (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265-275.

Stadtman, E. R. (1993). Oxidation of free Amino Acid residues in proteins by radiolosys and by metal-catalyzed reactions. Annual Reviews , 797-821.

Tang, Y., T. Shigematsu, S. Morimura, and K. Kida. (2005). Microbial community analysis of mesophilic anaerobic protein degradation process using bovine serum albumina (BSA)-fed continuous cultivation. J. Biosci. Bioeng. 99(2):150-164.

Negative

•Based in microscopic observations from the different positive enrichments, we found the same two microbial morphotypes present in all of the tubes that degraded collagen.

•We found that each environment responded to the collagen degradation differently; with respect to time intervals and lag phases that ranged from days to weeks.

•Only one environment tested did not degraded the collagen: Rice Paddy.

Positive

Enrichments

Serial Dilutions

Grass Rumen Fluid

Oxidation Lagoon (Coffee residues)

Residual Waters (Treatment Plant)

Environments Anaerobic collagen degraders

Kitchen oil trap 10 days Yes

Pig food 11 days Yes

Lucketti 18 days Yes

(coffee residue) 12 days Yes

Grass 16 days Yes

Rumen Fluid 15 days Yes

Residual water 13 days(Treatment plant)

Yes

Rice Paddy 55 days No