Characterizaon of the Microbial Fauna found in four local waterways of East Bridgewater, MA

Antonio Alves*, Windsor Lindor*, Que Chi Truong-Bolduc†, and Gilles Bolduc*

*Massasoit Community College, Brockton, MA 02302

INTRODUCTION Microbial Content RESULTS Watersheds are geographically disnct areas of land where surface water converges into a single point. As ground water is funneled through (Table 1) Nutrient, pH, and bacterial count data: the watershed it collects and spreads nutrients and pollutants across the Watershed chemical and bacteria results (June 2013 - August 2013) In order to characterize the microbial fauna found in our Nitrate Phosphate Salinity Total Bacteria Count Total Coliform Count Total Coliform Count Total E.coli Count watershed area. This study focuses on four bodies of water, in East Ph (ppm) (ppm) (ppt) (cfu/ml) (cfu/ml) EMB Plates (cfu/ml) MacConkey Plates (cfu/ml) EMB Plates water samples we performed the following procedures: Matfield River 0 Bridgewater , which are part of the Maield River sub-basin of the 0.2 1.25 6.74 2783 - 930 $ 117 5 45 watershed. The sub-basin contains some of the most (Farm) Poor Meadow Brook 0 0 - 0.4 0.275 - 0.75 6.87 - 7.15 160 0 - 30 2210 *** - 1680 $ densely developed areas in Massachuses, and has a recent history of ( Power Lines) 440 Total Colony & Robbins Pond site 1 0 - 0 0 - 0.6 0.475 - 1 5.84 - 6.47 100 0 - 0 125$ (<20) 45 not meeng EPA water quality standards. We sampled from Robbins (boat launch) Coliform Counts: Robbins Pond site 2 0 0.6 0.25 6.47 3740$ NA 210 0 Pond, Poor Meadow Brook, and Salisbury River to ( bridge) Salisbury river 2 0.2 1 6.88 5143 - 1,590 $ 702 30 characterize the microbial fauna, and determine their chemical and (Mattsfield Rd) 145 0 0.2 0.25 6.22 1933 - 755 $ 108 0 nutrient contents. We compared our findings with prisne waters -1 -2 Sawtuket River 70 We spread 0.1mL of water samples diluted 10 , and 10 , and Taunton River 0 - 0 0 - .06 0.275 - 1.25 6.75 - 7.13 875 - 105 - 3050$ 95 0 - 75 (Robbins Pond and Poor Meadow Brook) with those known to suffer undiluted on Trypc Soy Agar plates to determine the total (Rt 27) 455 from poor water quality (Salisbury and Maield Rivers). * EMB plate (gram negative), **10-2 dilution not included in average, *** undiluted sample not included in average, $- unfiltered number of bacteria /1 mL. We similarly plated samples on Eosin-Methylene Blue and MacConkey agar plates to Collection of Water Samples: determine the amount of coliforms and Escherichia coli/1ml. Dichotomous keys based on gram staining and biochemical test:

Isolaon of Bacterial GPS coordinates were recorded using an iPhone app, and water pH was Colonies: measured using standard test strips on site. Water was collected in three sterile 50 ml tubes. Addional water and mud was collected at Robbins Pond, Maield River and Poor Meadow Brook for Winogradsky columns. We isolated 51 bacterial isolates based on colony morphology. We recorded images of each isolate and performed a gram stain. Nutrient & pH test: Biochemical Test:

We performed the following biochemical test to aid in the Once in the lab we tested our water samples for pH Salinity, Nitrate, and classificaon of the 51 isolates: Phosphate levels. On-site pH readings were compared with a Meler pH meter. For the nutrient tests we used a LaMoe Direct Reading Titrator Citrate, Gelan Hydrolysis, Glucose fermentaon, Indole , Salinity Test Kit, LaMoe Low Range Comparator Phosphate Kit, and Lactose fermentaon, Methyl Red, NaCl, Nitrate Reducon, LaMoe Nitrate Nitrogen Tablet Kit. All our results fell within the Oxidase, Sucrose, Triple Sugar Iron, Voges – Proskauer acceptable ranges based on EPA criteria. The results were used to generate two dichotomous keys. CONCLUSION

ACKNOWLEDGEMENTS # of Bacteria Colonies This study was supported by: Matfield 4 • NSF-STEP Summer Research Program Winogradsky Poor MeadowBrook ( Power Lines) 11 Naonal Science Foundaon (NSF 06-502) Column and Wet Poor MeadowBrook ( Rt 27) 10 • n conjunction with i Mount: Robbins Pond 4 Students participated in the Massasoit Community College Undergraduate Research Experience SalisburyRiver 10 (SURE) program. Winogradsky Columns were constructed by filling 1 Liter Satucket 7 We would like to thank Prof. Michael Bankson for guiding us to the sites for water collection and boles with water and mud collected from Robbins Pond, Poor Retention pond (cranberry Bog) 5 providing us with information to the waterways. Meadow Brook, and Maield River. Wet mounts were diatoms algae protozoa desmids Matfield 2 0 6 6 We would also like to thank Prof. Marc Simmons for his help in identifying the eukaryotic examined under the microscope to idenfy non-bacterial Poor MeadowBrook ( Power Lines) 4 1 1 1 microorganisms. microorganisms present in the waterways. Robbins Pond 0 1 2 13