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Biol. Bull. 203: 251-252. (October 2002)
© 2002 Marine Biological Laboratory

Bacterioplankton Community Composition in Flowing Waters of the Ipswich River Watershed

Uri Y. Levine and Byron C. Crump

Ecosystems Center, Marine Biological Laboratory, Woods Hole, Massachusetts 02543
1 Department of Biology, Kenyon College, Gambier, OH 43022.

The Ipswich River (Massachusetts) watershed is the largest (404 km2), and most developed watershed of the three that compose the Plum Island Sound Long-Term Ecological Research (LTER) site. Twenty-five percent of the watershed has been set aside for conservation, but development is rapidly occurring in the southern portion of the watershed, because the area is attractive to Boston commuters (1). The effects of development upon a watershed are pronounced because land use is the most important factor dictating water quality, runoff, and productivity, and also contributes to other environmental problems (2). Past watershed studies have not characterized the natural bacterial community, focusing instead on overall microbial processes or the presence of enteric bacteria (3).

We hypothesized that land use would change the bacterioplankton community composition in the first-order streams (the smallest permanently flowing streams) of the watershed; so, for example, streams with predominantly urban land use would yield a unique microbial community. We also hypothesized that the communities of the first-order streams were likely to affect the population of the main stem of the river.

To investigate these hypotheses, we sampled first-order streams from within the watershed representing different land-use areas: urban streams 102 (78% urban land use) (labeling is consistent with that of the Ipswich-Parker Suburban Watershed Channel [IPSWATCH] [4]), 103 (90%), 104 (69%), and 161 (73%); forested streams 143 (81%) and 167 (56% forest and 27% urban); and agricultural streams 170 (28% agriculture, 37% forest, 13% urban) and 172 (28% agriculture, 15% forest, and 35% urban). Unfortunately, other primarily agricultural and forested sites could not be sampled, because the streams were either dry or not flowing as a result of the dry weather. The main stem of the river was also sampled at five points: at its intersection with Woburn St., Boston St. Bridge, Route 97, Winthrop St., and at the mouth of the river at the Ipswich River Dam (IRD, Sylvania Dam). The sites are visually represented in Figure 1A.



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  		Figure 1. (A) Map of the Ipswich watershed (modified from (4)) with the approximate location of sample sites. Sites along the main stem are labeled as Wob (Woburn St.), Bos (Boston St.), R97 (Route 97), Wnt (Winthrop St.), and IRD (Ipswich River Dam). (B) Multi-dimensional scaling diagram representing a pairwise similarity matrix (Dice) based on the presence or absence of DGGE bands (each band is assumed to represent an individual bacteria population) of bacterioplankton 16S rRNA genes taken from water samples collected during base flow conditions. Samples were collected on June 27, 2002, with the exception of I.S. 103, I.S. 104, Woburn St. (July), Boston St., Winthrop St., and IRD (July) which were collected on July 11, 2002. Circled populations represent waters either from similar land-use areas or from the main stem of the river that clustered together.

 
Base-flow water samples were collected on June 27 and July 11, 2002, in two acid-washed 1-l Nalgene bottles and kept on ice until further processing within 4 h of collection. Bacterioplankton were preserved for DNA extraction by passing as much as 1 l through 0.2-µm pore-size Sterivex filter capsules (Millipore) and flooding with DNA extraction buffer. Once concentrated on the filter, the samples were immediately frozen. The DNA extraction procedure was adapted from Zhou et al. (5). Polymerase chain reaction (PCR) products for denaturing gradient gel electrophoresis analysis (DGGE) were produced using 50 ng of each of the general bacterial primers 357f(g + c) (5'-CGCCCGCCGCGCCCCGCGCCCGGCCCGCCGCCCCCGCCCC-CCTACGGGAGGCAGCAG-3') (6) and 519r (ACCGCGGCTGCTGGCAC), and conditions adapted from Muyzer et al. (6). Approximately 50 µl of product, with 6x loading dye, was run on an 8% polyacrylamide gradient gel (30%–50% denaturant, 0.5x TAE) at 70 V for 17 h. Images were analyzed using GelCompar II and Statistica software package (StatSoft, Inc., Tulsa, OK).

With the exception of 161, all of the urban sites had relatively similar bacterial populations (Fig. 1B). Two streams with predominantly forested land-use percentages (167 and 170) clustered together despite 170 being classified as an agricultural site. Streams 161 and 172 also clustered together, presumably because 172 has more urban than agricultural land use. Stream 143, the other forested site, did not cluster with any other sites (data not shown). It is also notable that the sites which clustered together are from similar areas of watershed—102, 103, and 104 are all sites in Burlington, while 170, 167, 161, and 172 are located in the northeast part of the watershed.

The final grouping was that of the main stem of the river. Samples collected the previous summer from the Ipswich River Dam as well as a sample from the nearby Parker River Dam were analyzed with the current samples (data not shown), and it was found that the data from this summer are consistent with previously observed seasonal variations in bacterial community composition (Crump, unpubl. data). Interestingly, the Woburn St. samples had the most unique community compositions of the main stem samples, presumably because this site had more in common with the sites that are immediately upstream (102, 103, and 104).

In addition to indicating that different land-use areas produce distinct communities, the banding patterns also suggest that the bacterioplankton population of the river is established in the first-order catchments and is maintained throughout the course of the river. Of the 38 DGGE bands in the July Ipswich River Dam sample near the mouth of the river, 30 were found in first-order streams throughout the watershed.

In conclusion, we found evidence that microbial communities of first-order streams correlate with land-use type and control the composition of the bacterioplankton of the river.

The work was supported through the NSF grant OCE 976921.

Literature Cited

  1. PIE LTER website (http://ecosystems.mbl.edu/pie/data/wat/WATDescription.htm). Accessed 14 August 2002.
  2. Schneider, L. C., and R. G. Pontius. 2001. Agric. Ecosyst. Environ. 85: 83–94.
  3. Mallin, M. A., K. E. Williams, E. C. Esham, and R. P. Lowe. 2000. Ecol. Appl. 10: 1047–1056.
  4. IPSWATCH website (http://www.ipswatch.sr.unh.edu/). Accessed 14 August 2002.
  5. Zhou, J., M. A. Burns, and J. M. Tiedje. 1996. Appl. Environ. Microbiol. 62: 316–322.[Abstract]
  6. Muyzer, G., E. C de Waal., and A. G. Uitterlinden. 1993. Appl. Environ. Microbiol. 59: 695–700.[Abstract/Free Full Text]




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