This investigation evaluated the diversity of aquatic bacteria samples from Island Beach State Park, NJ. Barnegat Bay and estuarian pond samples were evaluated for nitrate, SRP phosphate, dissolved oxygen, and pH. Aquatic samples were filtered through 1.2 µm and 0.2 µm filters, genomic DNA extracted, amplified through PCR, cloned, and 16S rDNA gene sequenced for all isolates. A transformation rate of 34% was achieved within the two filter sizes and habitat subgroups tested. Colonies (n=344) were screened through electrophoresis for the 16S rDNA insert, clones (n=109) were selected for amplification and sequencing and evaluated through BLAST (n=33). These clones represented clones from 12 major groups. A phylogenic tree was constructed with the results. Eight groups were identified from the Barnegat Bay samples and 4 groups were identified from the estuarian pond samples.

INTRODUCTION

Traditionally microbial ecology studies have been limited to culturable bacteria and the characterization of diversity by biochemical testing. Within the last decade, a surge in genomic techniques have allowed advanced study of the unculturable world. Modern analysis of 16S rDNA from isolated microbial clones has begun a new era of molecular microbial ecological studies. The development of appropriate vectors and PCR primers can be used to study community complexities, species diversity, and evolutionary changes allowing scientists to better understand complex ecological processes (Stackebrandt, et al. 1995).
· The modern study of microbial diversity began with the pioneering work of Pace et al. Phylogenetic analysis of 16S ribosomal DNA (rDNA) clones obtained by PCR from uncultured bacteria inhabiting a wide range of environments has increased our knowledge of bacterial diversity (Pace, et al, 1995). The taxon composition of a microbial community is determined through analysis of 16S rDNA fragments generated by isolation and cloning processes (Frette, et al. 2003).
· The 16S rDNA is the gene of interest because: 1) it is comprised of highly conserved and hypervariable regions; 2) it is found in all living organisms; 3) it acts as a molecular clock correlating to changes in evolutionary time (Lemke, 2003).

OBJECTIVES

This investigation evaluated the following questions:
1) Are there any differences in bacterial diversity due to habitat heterogeneity?
2) What is the relationship among the bacteria taxa isolated from the sample sites?

HYPOTHESIS
It is hypothesized that the estuarian pond site tested offers a greater heterogeneity in habitat and therefore greater species diversity when compared to the Barnegat Bay site.