Hixon 1 Concentrations and identities of fecal coliform bacteria from ...

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Hixon 6 NAME OF SAMPLE AREA LB1 DR1 JU2 JU1 ARL7 ARL3 LOCATION Long Branch at Wigmore Street Deer Creek at Talleyrand Avenue JU boathouse dock Creek/pipe runoff at JU dock inlet Unnamed Stream at Ferber Road Red Bay Branch at Lone Star Road Figure 1: Locations of Sample sites. COJ does not monitor JU1 and JU2. http://www.coj.net/NR/rdonlyres/ejld2jez3myhqoq37l2wi4c2rsf2paanfw5s4tqsa4zyg4ws2wkrhy q6w5bmj27lvb7aihpyupfxmve6kd3om3yslkc/Fecal+map+1st+quarter+2004.pdf At each sample site, an 800mL water sample was collected in a sterile plastic container just prior to low tide. Water temperature, salinity, and dissolved oxygen were measured in a small separate sample collected in a 50mL beaker using Exstik DO600 Dissolved Oxygen meter and Exstik pH/ Conductivity/ TDS/ Salinity/ Temperature meter. All samples were taken back to the Jacksonville University microbiology lab for testing no later than 4 hours after collection.
Hixon 7 Testing began with using the membrane filtration technique: triplicates of 50mL of each sample was filtered on 0.45µm filters and then placed onto M-FC agar (Fisher Scientific Catalog Number DF0677173), which is selectable for fecal coliform bacterial growth, in 50x9mm Petri dishes. The incubation period for the 1/29/11 collection was 24 hours and 37 minutes and 18 hours and 35 minutes for 3/12/11 collection at 46 ºC. Fecal coliform CFUs were visually counted using a Leica Quebec Darkfield Colony Counter and these numbers were multiplied by two to calculate fecal coliform CFUs per 100mL (table 1). Only blue colonies were counted, all brown or off white colonies were considered not to be fecal coliform bacteria CFUs. Water pH was measured using a Corning pH meter 125, which was calibrated prior to use. To calculate total bacterial counts, triplicates of serial dilutions using 0.85% NaCl in 100mL dilution containers were completed in tenfold increments from tenfold to one-thousand fold, followed by plating a 0.1mL sample onto TSA and incubating for 42 hours and 5 minutes at 37 ºC prior to counting. IDENTIFICATION OF FECAL COLIFORM BACTERIA Morphological, physiological, and metabolic tests were used to identify and differentiate fecal coliform bacteria from each of the sampling locations. Three random and separate fecal coliform bacteria colonies, one from each of the three triplicate plates per sample site (totaling 18 isolates), were chosen and successively quadrant streaked six times on m-endo agar (Fisher Scientific Catalog Number DF0677173), in order to promote only fecal coliform bacteria growth. Single colonies were used for testing and each sample was quadrant streaked on a new m-endo plate weekly in order to have living bacteria available for all tests. Before individual fecal coliform bacteria testing for differentiation among fecal coliform bacteria began, m-endo slants were streaked, incubated for 11 hours and 20 minutes, sealed tight, and store in a refrigerator with a temperature ranging from 0 to 5ºC for preservation. The methods for identification were
Page 1 and 2: Hixon 1 Concentrations and identiti
Page 3 and 4: Hixon 3 Fecal coliform bacteria are
Page 5: Hixon 5 levels, fecal coliform bact
Page 9 and 10: Hixon 9 All samples were collected
Page 11 and 12: Hixon 11 573,300 LB1 1/29/2011 TS 1
Page 13 and 14: Hixon 13 bacteria concentrations. A
Page 15 and 16: Hixon 15 Figure 4: Micrograph of Es
Page 17 and 18: Hixon 17 SAMPLE / GENUS DR1 BACTERI
Page 19 and 20: Hixon 19 SAMPLE / GENUS SIZE (µm)
Page 21: Hixon 21 Servias P, Passerat J. 200

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