Attached Algae as an Indicator of Water Quality: A Study of the Viability of Genomic Taxonomic Methods

Transcription

1 Attached Algae as an Indicatr f Water Quality: A Study f the Viability f Genmic Taxnmic Methds Allisn Wd, Senir, University f New Hampshire Alisn Watts, PhD, University f New Hampshire

2 Special Thanks: Paul Stacey, Great Bay Natinal Estuarine Research Reserve Jeff Hall, Kelley Thmas and the Hubbard Genme Center US EPA Greater Research Opprtunities Fellwship Prgram

3 Outline: Prject Gals Backgrund Sampling Methdlgy Data Analysis & Results Next Steps

4 Prject Gals #1 Verify that algae cnstitute a viable ecsystem indicatr fr Great Bay #2 Cmpare traditinal micrscpic methds t genmic analysis Micrscpe $450/sample (100-1,000 species) Genmic $50/sample (100,000 species) #3 Further explre genmic methds

5 Why Algae? Sensitive t nutrient enrichment Shrt life cycles, react quickly Variety f species Dr. Ruth Patrick rudcenter.r g/newsletters /2013/issue6/ img/ruthpatrick.jpg Ppulatin density Estuarine & freshwater Easy t cllect hhttp://static01.nyt.cm/images/2013/09/24/us/patric k-bit2/patrick-bit2-master675.jpg

6 Current Applicatins: United States Gelgical Survey (USGS) Prter et al., 2008 Natinal database f algal - metric indicatrs Including reginal indicatrs Categries include: Trphic cnditin, Salinity, Chlride, Organic enrichment, ph, Calcium, etc.

7 Current Applicatins: State mnitring prgrams Maine Dept. f Envirnmental Prtectin Bimnitring Prgram Develped mdel fr assessment using specific indicatrs fr Maine ecsystems with success TP, TN, % Impervius cver, etc. Danielsn et al., 2011

8 Sampling Methd Periphytmeter Maine DEP: Surface Cllectin Cntrlled fr substrate, time, light, flw, depth Glass slides submerged fr 2-week intervals Great Bay: Periphytmeter

9 Sample Sites Exeter, Squamsctt and Lamprey Rivers; Great Bay Attached algae Water chemistry Nutrient cntent Lamprey River Great Bay Squamsctt River Exeter River

10 A N A L Y S I S

11 Gal #1: Traditinal Micrscpic Analysis Algae were identified and % abundance f each species was btained frm each sample site Prter et. al, USGS, Algae classificatins (mstly diatms) Species classified by attributes; Nutrients, ph, salinity, etc. Analyzed in cmbinatin with site data Principal cmpnents analysis in JMP

12 Sites 3 & 7 had HIGH Phsphrus Levels Principal cmpnents analysis in JMP Pink sites had greater than 75 µg/l TP

13 Gal # 2: Genmic Analysis Cmparisn Micrscpe methds Searching fr knwn target Genmic methds Can identify hundreds f thusands f individual rganisms, even in small samples Unless we can match that DNA/RNA t a knwn database, can t identify species Full DNA sequencing Technlgy available, very expensive / time cnsuming Barcding Identifies species based n nly a small segment f DNA sequence

14 Methd: Next-Generatin Sequencing Shrt pieces f DNA/RNA are washed acrss a flw cell with selected primers Thse that stick are amplified, frming clusters Strands are tagged with indicatrs ne base at a time Indicatrs are hit with a laser, activating a crrespnding flurescent clr and read with a camera Prduces millins f highly accurate reads

15 Challenges thus far (Gal # 2) Algae database are incmplete Results nt as specific as micrscpe Unable t reslve mst taxa past family, we need species t relate t indicatrs Nt sure if database issue r methd issue DNA/RNA segment might nt prvide enugh detail NO DIRECT COMPARISON POSSIBLE CURRENTLY BUT we have thusands f ther micrrganisms, such as BACTERIA...

16 Gal # 3: Further explre genmic analysis Using the genmic data Trying t identify ther ptential indicatrs frm the micrrganism infrmatin we d have Such as BACTERIA

17 Bacterial Results Pink triangles are bth upstream sites w/ very high Phsphrus Blue circles are sites dwnstream in the tidal rivers Black squares are sites in Great Bay

18 Next Steps Research Determine if there are additinal reference database fr algae samples/decide whether t build ur wn Identify additinal indicatr species frm genmic data Regulatin Determine if this methd hlds value fr NH stream mnitring prgrams Early indicatin f changes in water quality Targeted management effrts

19 Questins?