Pelican and Fish Mortality Report Final

City of Saint Petersburg
TECHNICAL MOMRANDUM
Water Quality Sampling
Fish and Pelican Mortality Incident of January 2017
March 2017
TECHNICAL MEMORANDUM
TECHNICAL
MOMORANDUM
Water Quality Sampling Fish/Pelican
Kill of January 2017
Prepared for:
Carlos Frey PE
Scott Lehman Senior Engineer
Project Manager
One 4th Street North
Saint Petersburg FL, 33704
Prepared by:
Arcadis U.S., Inc.

3109 West Dr. Martin Luther King Jr.
Boulevard
Stephen Rice Suite 350
Senior Ecologist Tampa
Florida 33607
Tel 813 903 3100
Fax 813 903 9115
Our Ref.:
01814066.0000
Date:
March 2017
This document is intended only for the use of

the individual or entity for which it was
prepared and may contain information that is
privileged, confidential and exempt from
disclosure under applicable law. Any
dissemination, distribution or copying of this
document is strictly prohibited.
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CONTENTS
1 Executive Summary .............................................................................................................................1-1
2 Background ..........................................................................................................................................2-1
3 Scope ...................................................................................................................................................3-2
4 Sample Collection and Analysis...........................................................................................................4-2
Sample Collection ................................................................................................................................4-2

January Analysis ..................................................................................................................................4-3
March Analysis .....................................................................................................................................4-3
5 Conclusions and Recommendations ...................................................................................................5-4
Conclusions..........................................................................................................................................5-4
Graphic 1: AccuWeather Temperatures for Saint Petersburg .........................................5-4
Recommendations ...............................................................................................................................5-6
6 References ...........................................................................................................................................6-8
TABLES
Table 1. Water Quality Parameters January 19, 2017 .............................. Error! Bookmark not defined.
Table 2. Analaytical Results January 19, 2017 ......................................... Error! Bookmark not defined.
Table 3. Water Quality Parameters March 6, 2017 ................................... Error! Bookmark not defined.
Table 4. Analaytical Results March 6, 2017 .............................................. Error! Bookmark not defined.
FIGURES
Figure 1. Location Map ................................................................................. Error! Bookmark not defined.
Figure 2. FWC Mortality Locations
Figure 3. January Sampling Locations
Figure 4. March Sampling Locations
APPENDICES
Appencix A - Theory Graphics
Appendix B FWC Report
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Appendix C AEL January Sample Report

Appendix D Source Molecular Sample Report
Appendix E AEL March Sample Report
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1 EXECUTIVE SUMMARY
On January 10, 2017, the City of Saint Petersburg (City) received a report of a fish kill at the residential
stormwater lake, Riviera Lake #1 adjacent to Riviera Bay (Figure 1). Additionally, during the period from
January 11th through 30th, 2017, reports of 70 dead or ill pelicans were received by the Florida Fish and
Wildlife Conservation Commission (FWC) at locations in and around Saint Petersburg. Arcadis was
retained by the City of St. Petersburg to investigate the potential causes of the pelican deaths and fish kill.
The original scope of the investigation, as determined by Arcadis, was to collect surface water samples at
Riviera Lake #1 and Coffee Pot Bayou and collect other records that may be pertinent to the
investigation. Surface water samples were submitted for laboratory analysis for fecal coliform, fecal
speciation, nitrogen, phosphorus, and other nutrients. An additional surface water sampling event at
Riviera Lake #1 was deemed necessary. Additional water samples were collected on March 6, 2017 to
confirm the January 19, 2017 analytical results and suspected thermal stratification of the Lake.
Based on the surface water analytical results and water quality parameter data collected on January 19
and March 6, 2017, the most probable explanation of the pelican deaths and fish kill was the
eutrophication of Riviera Lake #1 brought on by a thermal inversion of the stratified layers of the Lake
coupled with an outbreak of Type E botulism toxin. A sudden drop in atmospheric temperature and cool
rain occurred between January 8th and January 10, 2017, which disturbed the thermal stratification of the
Lake and provided for mixing of surface and deep water layers. Eutrophication is caused by excessive
amounts of nitrates and phosphates and stimulates an explosive growth of algae which depletes the
water of oxygen. Algae became infected by a rare bacteria, Clostridium botulinum, produced under the
low-oxygen conditions at Rivera Lake #1. Clostridium botulinum produces Botulinum toxins thought to
have further contributed to the fish kill. Pelicans ingested the infected fish at Riviera Lake #1 and became
ill or died. See Appendix A for a graphical illustration of the thermal inversion scenario offered as the
impetus for the zoonotic infection (botulism) outbreak. Pelican deaths noted in other locations likely
ingested infected fish at Riviera Lake #1.
Arcadis offers recommendations in Section 5.0 to reduce the potential for another fish kill event at Riviera
Lake #1.
2 BACKGROUND
On January 10, 2017, the City of Saint Petersburg (City) received report from a homeowner located
adjacent to Riviera Lake #1 indicating thousands of dead fish were attracting vultures. The Citys
Stormwater Maintenance staff responded to the incident and removed approximately 5,000 pounds of
dead fish from Riviera Lake #1. On January 11, 2017, City employees and private citizens began
reporting dead or ill pelicans in Riviera Lake #1 and surrounding areas. Reports were received by the
Florida Fish and Wildlife Conservation Commission (FWC). During the period from January 11th through
30th, 2017, reports of 70 dead or ill pelicans were received by the FWC (Appendix B). The approximate
locations of these reports were included in their report (Figure 2).
As of the issuance of this Technical Memorandum, a final report with the total number of dead pelicans
and results of pelican necropsies attributed to this incident are not yet available from FWC. The following
sections detail surface water sample collection events, analytical results, and offer conclusions and
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recommendations developed by Arcadis. This Technical Memorandum offers an explanation of the fish
kill and pelican deaths, based on the best available data and science.
3 SCOPE
On January 16, 2017, Arcadis was retained by the City to investigate the pelican deaths and fish kill. The
City broadly defined the scope of work as a science-based investigation and development of a theory as
to the cause(s) of the pelican deaths and fish kill. In response, Arcadis developed the following scope of
work based on the Citys request:
Collect surface water samples and water quality readings at Coffee Pot Bayou and Riviera Lake
#1.
Analysis of surface water samples for fecal coliform, fecal speciation (human, dog or bird),
nitrogen, phosphorous, and other nutrients.
Obtain pertinent information to aid in the investigation. Requested information included:
o Historic fish kill incident reports (if any)
o Geographic Information System (GIS) data depicting the locations of the reclaimed water
pipes in Riviera Lake #1
o Residual fecal coliform concentrations from daily samples collected at the Citys reclaim
Waste Water Treatment System systems
o Data collected from FWC investigations of the incidents.
The scope was broadened to allow for an additional surface water sample collection event and water
column investigation at Riviera Lake #1 on March 6, 2017. The additional sampling event was
deemed necessary, following the receipt of January 19, 2017 analytical results, in order to confirm
elevated lake nutrient levels and suspected thermal stratification. The following section summarizes
the sample collection efforts and laboratory analytical results.
4 SAMPLE COLLECTION AND ANALYSIS
Sample Collection
On January 19, 2017, Arcadis mobilized to collect surface water samples and water quality data at three
locations at Riviera Lake #1 and seven locations at Coffee Pot Bayou. Figure 3 depicts the January 19,
2017 sample locations. Surface water samples were collected at a depth of six inches below the water
surface. Water quality measurements collected at the surface on January 19, 2017 included: conductivity,
dissolved oxygen, pH, and temperature.
The surface water samples from Riviera Lake #1 and Coffee Pot Bayou were submitted to Advanced
Environmental Laboratory Inc. (AEL) in Tampa, FL for analysis of: ammonia, chlorophyll a, chlorophyll b,
chlorophyll c, coliform fecal, coliform total, nitrate, nitrite, pheophytin, total dissolved solids, total kjeldahl
nitrogen, total phosphorus, and total suspended solids. Surface water samples were also sent to Source
Molecular laboratory in Miami, FL where they were prepared and held for future fecal coliform speciation
to determine the coliform contribution (human, dog or bird). Three samples with the highest concentration
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of fecal coliform were sent for speciation: T1701119lk-01 from Riviera Lake #1, and T170119-01 and
T170119-6 from Coffee Pot Bayou (Figure 3).
January Analysis
Analytical results received from the January 19, 2017 sampling event indicated eutrophic conditions at
Riviera Lake #1. Eutrophication is caused by excessive amounts of nitrates and phosphates and
stimulates an explosive growth of algae and depletes the water of oxygen. Algae became consumed by
bacteria at Rivera Lake #1. Table 1 presents the water quality parameters collected at Riviera Lake #1
on January 19, 2017. Table 2 presents the analytical data collected at Riviera Lake #1 on January 19,
2017. Table 2, shows elevated algal concentrations as evidenced by increased chlorophyll a, chlorophyll
b, chlorophyll c, pheophytin a, coliform fecal, and coliform total. The presence of chlorophyll a is linked to
the presence of cyanobacteria, and at the reported concentrations indicate algae bloom conditions.
Eutrophic conditions are also evidenced by increased ammonia, nitrate, nitrite, total kjeldahl nitrogen, and
total phosphorus.
In the sample submitted for coliform speciation (T170119lk-01), concentrations of human fecal coliform
were low, bird fecal coliform was detected in quantities below the limit of quantification, and dog fecal
coliform was not detected. Appendix C includes the January 19, 2017 analytical results as reported by
AEL. Appendix D includes the fecal contribution (human, dog or bird) as reported by Source Molecular.
Analytical results received from the January 19, 2017 sampling event at Coffeepot Bayou indicated typical
tidal inlet conditions, with the exception of one sample location (T170119-01) (Figure 3). The value
reported for dissolved oxygen at location T170119-01 (6.3 milligrams per liter [mg/L]) was lower than the
typical range (7.5 9.6 mg/L). This location corresponds with the highest values for nitrogen,
phosphorous, fecal coliform, and total coliform. The sample was submitted for coliform speciation,
concentrations of human fecal coliform were low, bird fecal coliform was detected in quantities below the
limit of quantification and dog fecal coliform was not detected. A second sample (T170119-06) from
Coffee Pot Bayou was submitted for coliform speciation. Concentrations of human and dog fecal coliform
were not detected, bird fecal coliform was detected in quantities below the limit of quantification. Appendix
C includes the January 19, 2017 analytical results as reported by AEL. Appendix D includes the fecal
contribution (human, dog or bird) as reported by Source Molecular.
March Analysis
Based on the results of the January 19, 2017 sample collection event, additional water quality profiling
was performed at Riviera Lake #1 to confirm the eutrophic conditions and a suspected thermocline. A
thermocline is a steep temperature gradient in a body of water, marked by a distinct layer between the
mixed layer at the surface and the deep water layer. On March 6, 2017 Arcadis mobilized to collect four
surface water samples and record water quality parameters to the 26 foot depth at five water column
locations. Figure 4 depicts the March 6, 2017 sample locations at Riviera Lake #1. The analytical results
and water column profile data confirmed eutrophic conditions as evidenced by elevated nutrient levels
and confirmed a thermocline as evidenced by increased salinity and decreased dissolved oxygen,
temperature, and pH below the 6 to 8 feet water level layer at Riviera Lake #1. On March 6, 2017,
salinity/density stratification was noted in water quality parameters collected at Riviera Lake #1, beginning
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at the 6 to 8 ft depth interval. Dissolved oxygen levels were observed to be low (anoxic) below the 6 to 8
ft depth (the hypolimnion). The upper 6 ft of the water column (the epilimnion) is the primary aerobic zone
where fish and pelicans feed.
Table 3 presents the water quality parameters at two foot intervals to the lake bottom (26 feet) and Table
4 presents the analytical data collected at Riviera Lake #1 on March 6. 2017. The surface water samples
were submitted to AEL for the same analysis as was completed on April 19, 2017. Appendix E includes
the March 6, 2017 analytical results as reported by AEL. The analytical results from the March 6, 2017
sampling event confirm the high nutrients levels noted in January 19, 2017 analytical data.
5 CONCLUSIONS AND RECOMMENDATIONS
Conclusions
Based on the surface water analytical results and water quality parameter data collected on January 19
and March 6, 2017, the following conclusions are offered for the most probable explanation of the pelican
deaths and fish kill incidents.
Depth to base measurements indicate Riviera Lake #1 is a deep costal storm water lake, with the deepest
point measured at 26 ft below surface. The Lake has brackish water quality and bottom sediments
consisting of a thick anaerobic organic layer over sand. Nutrient loads in this urban lake are high, likely
due to reclaim water in puts, fecal sources and potential runoff of lawn fertilizers. Lake water
temperatures are relatively high year-round with weak thermal stratification present during the winter
months.
Predominant warm weather conditions were interrupted on January 8, 2017 when temperatures dropped
from an average high of 70 degrees Fahrenheit (F) to a day high of 52 degrees F and then rebounded
back to 70 degrees by January 10, 2017 as shown in the Graphic 1 below
(http://www.accuweather.com/en/us/st-petersburg-fl/33712/january-weather/332287). Additionally, a cool
rain event on January 10, 2017 ended an extended period of no rain.
Graphic 1: AccuWeather Temperatures for Saint Petersburg
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Cool, infrequent rains and air temperatures experienced prior to January 10, 2017, disturbed the
thermocline at Riviera Lake #1 providing for mixing of hypolimnion and epilimnion. This was evidenced
by brown color of algae suspended in water column in documented in photographs and video on January
19, 2017.
Eutrophication of Riviera Lake #1 caused by excessive amounts of nitrates and phosphates and
stimulated an explosive growth of algae that depleted the water of oxygen. The algae were infected by
bacteria, including Clostridium botulinum, produced under the low-oxygen conditions. Botulinum toxins
produced by the bacteria are the suspected cause of the zoonotic infection (botulism) outbreak. As anoxic
conditions mix through the water column, following a thermal inversion, fish begin to ingest infected algae
and die from both the anoxic water and the toxic effects of the bacteria. The dead fish provide a
continuous source of protein and the bacteria continue to reproduce, further contributing to fish kills. The
Citys response to the fish kill was prompt; however pelicans began ingesting infected fish shortly after
they died and floated to the surface at Riviera Lake #1. Pelicans died or were symptomatic after
ingesting botulism-infected fish. Avian symptoms reported were consistent with those due to toxin
ingestion; degrading motor functions of the head, neck and wings but retained ability to use legs to walk
or paddle. See Appendix A for a graphical illustration of the thermal inversion scenario offered as the
impetus for the zoonotic infection (botulism) outbreak.
Appendix A graphically illustrates Arcadiss theory of lake eutrophication and zoonotic infection (botulism)
as the likely source of the January 2017 fish kill incident at Riviera Lake # 1, and subsequent pelican
deaths.
Zoonotic infection (Clostridium botulinum), present in hypolimnion, reproduces in systems with low
dissolved oxygen and high algal detritus. Botulism in coastal waters (higher saline) generally consists of
two types: C & E.
Type C is the most common serum affecting avian waterfowl because it is generally disseminated
through the invertebrate population based food web. Invertebrates are immune and therefore
bioconcentrate the Type C toxins. Affected ecological populations would consist of small to large
birds who primarily feed on invertebrates, fish or other carcasses with large invertebrate (i.e, fly
maggots).
Type E is a more common in higher saline (coastal) waters and generally is transmitted in an
algae/planktonic sourced food web. Thus, larger planktonic and algae feeding fish would be a
vector to primarily larger fish eating birds, at the onset. If dead fish are not removed from
waterbody then a Type C scenario can develop.
Salinity levels at Riviera Lake #1 are brackish and the bacteria would more closely align with Type E
Botulinum toxin under these conditions. Type E botulism affect larger algae eating fish with toxic doses of
bacteria. The references cited in this memorandum conclude that this plankton- based and type of
botulism is reported in the gut contents of tilapia. Species of fish recovered were reported by the City as
mullet and tilapia. Mullet primarily feed on invertebrates, fish and macroalgae. Tilapia feed on planktonic
and macroalgae.
Smaller, invertebrate-eating birds, are unaffected due to lack of time of the Type E Botulinum toxin to
enter and bioconcentrate in the invertebrate food web. However, larger birds, like pelicans, die or are
symptomatic after eating infected fish. Avian symptoms reported during January 2017 were consistent
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with toxin ingestion including: degrading motor functions of the head, neck and wings but retained abilities
to use legs to walk or paddle.
Planktonic algae appear to be the vector with tilapia, concentrating Botulism toxin (Type E) in their gut.
However, it should be noted that it has not been confirmed that the bacteria exist in Riviera Lake #1, due
to the lack of available live tissue to test. Dead pelicans were sent for necropsy; however as of the
issuance of this memorandum, the necropsy results are not available from FWC and the University of
Florida. Available research does not pinpoint all conditions needed to produce blooms of clostridium but
relatively high temperature, low dissolved oxygen and available decaying matter from plant or animal are
all necessary. If a botulism outbreak did occur as suspected, it should be noted that as thermal
stratification is re-established at Riviera Lake # 1 and algae growth again increases, a change in water
color from brown to green will be observed. Subsequent algae blooms supersaturate the epilimnion with
oxygen and halt bacterial toxin production in the epilimnion. However, production of bacterial clostridium
spores permits survival in a dormant state until the spores are exposed to favorable conditions again (i.e.,
low dissolved oxygen, decaying protein sources etc.).
Anecdotal observations of previous fish kills at Riviera Lake #1 were made know to Arcadis through this
investigation. Arcadis offers that previous fish kills have likely resulted as part of yearly increased diatom
and planktonic algal bloom and subsequent collapses at Riviera Lake #1.
Pelicans removed from Coffee Pot Bayou may not be related to event at Riviera Lake #1; however some
pelicans removed from this area were sent to labs for pathology/necropsy. It is conceivable that pelicans
who fed on fish at Riviera Bay Lake #1 flew back to rookery at Coffee Pot Bayou prior to becoming
symptomatic and dying. All avian and mammalian (one dog recovered in Coffee Pot Bayou) specimens
were removed and have been sent to lab for pathology. As of the issuance of this memorandum,
necropsy results are not yet available from FWC and the University of Florida.
Recommendations
Arcadis offers the following recommendations to eliminate or reduce the potential for another fish kill
event at Riviera Lake #1:
Eutrophic conditions should be studied, and phosphorous levels reduced at Riviera Lake #1.
Investigate the reclaim water utility inputs to Riviera Lake #1 and eliminate any entry points of the
reclaim water into the Lake.
Aerators should be installed at Riviera Lake #1 to increase dissolved oxygen throughout the
water column, especially during times of mixing with hypolimnion (winter rain events), as the
Clostridium botulinum bacteria only reproduce in anaerobic conditions.
Riviera Lake #1 has likely experienced a Clostridium botulium bacterial outbreak, therefore
remnant bacterial spores exist at the Lake and are available to reproduce during future favorable
conditions. As such incident response coordination between the City of St. Petersburg and FWC
should be formalized to define responsibilities for impaired live fish collection efforts and water
sampling. Predetermined actions and responsibilities for live fish collection, water sample
handling, time frames, shipping, and contracted laboratories should be established. For example,
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impaired live fish, invertebrates or other wildlife should be collected and tested immediately for
Clostridium botulium.
Incident response and removal of dead fish should continue to be prompt due to the risk of
contributing to a Type C botulism outbreak. A Type C botulism outbreak would affect many more
species and likely result in thousands of birds deaths within a relatively short time frame.
Formal record keeping, documenting incident response to fish kills within the City of St.
Petersburg should exist in a centralized web platform to allow for greater public access.
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6 REFERENCES
Lafrancois, B.M., Riley, S.C., Blehert, D.S., and Ballmann, A.E. 2011. Links between type E
botulism outbreaks, lake levels, and surface water temperatures in Lake Michigan, 1963-
2008. Journal of Great Lakes Research. 37. pp. 86-91.
Lalitha, K. V. and Gopakumar, K. Sensitivity of Tilapia (Oreochromis mossambicus) to
Clostridium botulinum toxins. Aquaculture Research. 2001. 32. pp. 761-764.
Nol, P.; Rocke, T. E. ; Gross, K.; and Yuill, T. M. Prevalence Of Neurotoxic Clostridium
Botulinum Type C In The Gastrointestinal Tracts Of Tilapia (Oreochromis Mossambicus) In
The Salton Sea. Journal of Wildlife Diseases, 40(3), 2004, pp. 414419.
Nol, P.; Williamson, J.L.; Rocke, T. E. ; and Yuill, T. M. Prevalence Of Neurotoxic Clostridium
Botulinum Type C In The Gastrointestinal Tracts Of Tilapia (Oreochromis Mossambicus) In
The Salton Sea. Journal of Wildlife Diseases, 40(3), 2004, pp. 414419.
U.S. Environmental Protection Agency. Type E Botulism Outbreaks: A Manual for Beach
Managers and the Public. Great Lakes National Program Office (GLNPO)
U.S. Geological Survey (USGS). 1999. Avian Botulism, In Field Manual of Wildlife
Diseases: General Field Procedures and Diseases of Birds, pp. 271-282.
http://www.nwhc.usgs.gov/publications/field_manual/chapter_38.pdf.
USGS National Wildlife Health Center. 2006. Disease Information, Avian Botulism.
November 7. http://www.nwhc.usgs.gov/disease_information/avian_botulism/index.jsp.
USGS National Wildlife Health Center. 2007. Avian Botulism. Disease Information.
http://www.nwhc.usgs.gov/disease_information/avian_botulism/index.
Yule, A. M.; Barker, I. K.; Austin, J. W.; and Moccia, R. D. Toxicity of Clostridium Botulinum
Type E Neurotoxin To Great Lakes Fish: Implications For Avian Botulism. Wildlife Disease
Association 2006. Journal of Wildlife Diseases, 42(3), 2006, pp. 479493.
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TABLES
Table1WaterQualityParametersJanuary2017
CoffeePotBayou
(January19th2017)
2
Location TotalDepth(ft) AnalysisDepth pH Temp(C) Conductivity(S/cm ) DisolvedOxygen(mg/L)
T17011901 1.10 Surface 7.07 21.98 37,465 6.33
T17011902 7.00 Surface 7.98 22.1 37,693 9.62
T17011903 9.20 Surface 8.1 21 37,352 8.63
T17011904 8.14 Surface 8.14 21.28 37,520 9.06
T17011905 9.10 Surface 8.13 21.9 37,901 8.66
T17011906 8.10 Surface 8.11 21.77 38,175 8.08
T17011907 6.90 Surface 8.09 20.88 39,023 7.54
RivieraLake#1
(January19th2017)
2
Location TotalDepth(ft) AnalysisDepth pH Temp(C) Conductivity(S/cm ) DisolvedOxygen(mg/L)
T170119LK01 >2 Surface 8.06 21.8 17,533 10.83
T170119LK02 >2 Surface 8.25 23.02 17,583 12.33
T170119LK03 >2 Surface 8.45 25.26 17,740 15.15
Table2January2017LaboratoryResults
LaboratoryResults
January19th2017SamplingEvent
Ammonia(N) ChlorophyllA ChlorophyllB ChlorophyllC Coliform Coliform NitrateasN NitriteasN PheophytinA Total TotalKjeldahl Total Total
SAMPLEID (mg/L) (mg/m3) (mg/m3) (mg/m3) Fecal(#/100 Total(#/100 (mg/L) (mg/L) (mg/m3) Dissolved Nitrogen Phosphorus Suspended
T17011901 0.02U 9.1 1U 1U 918B 1320B 0.18U 0.18U 1U 26000 0.46 0.109 15
T17011902 0.02U 5.1 1U 1U 1U 20B 0.18U 0.18U 1.6 26000 0.3 0.089 14
T17011903 0.02U 8 1U 1U 10B 20B 0.18U 0.18U 1.7 26000 0.26 0.008I 15
T17011904 0.02U 6.9 1U 1.5 20B 30B 0.18U 0.18U 2.1 27000 0.22 0.084 14
T17011905 0.02U 10 1U 1.5 1U 10B 0.18U 0.18U 1U 27000 0.3 0.094 15
T17011906 0.02U 9.6 1.2 1.2 1U 30B 0.18U 0.18U 1U 27000 0.5 0.097 13
T17011907 0.02U 5.7 1U 1U 1U 1U 0.18U 0.18U 1U 28000 0.4 0.092 13
T170119lk01 0.71 67 12 4.4 64B 118B 0.18U 0.18U 3.2 12000 3.1 0.416 24
T170119lk02 0.3 85 15 7 1U 118B 0.18U 0.18U 1.3 12000 3.1 0.381 34
T170119lk03 0.1 76 13 11 1U 60B 0.18U 0.18U 1U 12000 3.1 0.394 34
Qualifiers
UThecompoundwasanalyzedforbutnotdetected
IThereportedvalueisbetweenthelaboratorymethoddetectionlimitandlaboratoryp
BResultsbaseduponcolonycountsoutsidetheacceptablerange.
Table3WaterQualityParametersMarch2017
RivieraLake#1Location(T70306LK1)
(March6th2017)
AnalysisDepth pH Temp(C) Conductivity(S/cm2) DisolvedOxygen Turbidity OxidationReductionPotential(g/mv)
Surface 7.45 20.25 25,096 2.00 21.75 149.5
2ft 4.42 20.21 25,095 1.75 19.13 161.3
4ft 7.38 20.2 25,105 1.56 19.36 177.4
6ft 7.06 20.27 25,203 0.51 54.14 226.9
8ft 6.91 20.11 28,200 0.06 8.74 302.6
10ft 6.91 19.83 28,380 0.07 5.96 300.5
12ft 6.92 19.65 28,354 0.07 6.68 297.2
14ft 6.93 19.65 28,375 0.08 9.38 292.3
RivieraLake#1Location(T70306LK2)
(March6th2017)
Surface 7.5 20.4 25,171 2.39 23.99 114.2
2ft 7.49 20.37 25,126 2.34 24.55 123.1
4ft 7.46 20.31 25,127 2.09 26.17 151.5
6ft 7.42 20.3 25,123 2.08 27.43 178.4
8ft 6.91 20.26 25,189 1.59 29.3 235.5
10ft 6.89 19.73 28,202 0.07 10.7 311.5
12ft 6.89 19.67 28,263 0.07 10.01 309.6
14ft 6.89 19.67 28,253 0.07 10.8 306.2
16ft 6.9 19.71 28,337 0.07 95.13 301.3
RivieraLake#1Location3(T70306LK3)
(March6th2017)
Surface 7.52 20.68 25,179 3.09 33.46 110.9
2ft 7.53 20.63 25,135 2.62 36.66 122.4
4ft 7.51 20.56 15,145 2.41 40.98 137.8
6ft 7.47 20.51 25,165 2.33 43.12 170.4
8ft 6.95 20.34 27,470 0.47 38.93 279.8
10ft 6.91 19.8 28,193 0.07 19.58 295.9
12ft 6.92 19.75 28,209 0.07 17.41 289.8
Table4March2017LaboratoryResults
LaboratoryResults
March6thSamplingEvent
Ammonia(N) ChlorophyllA ChlorophyllB ChlorophyllC Fecal(#/100 Total(#/100 NitrateasN NitriteasN PheophytinA Dissolved Nitrogen Phosphorus Suspended
SAMPLEID (mg/L) (mg/m3) (mg/m3) (mg/m3) mL) mL) (mg/L) (mg/L) (mg/m3) Solids(mg/L) (mg/L) (asP)(mg/L) Solids(mg/L)
T170306LK1 0.16 81 90 9.3 20B 200 0.18U 0.18U 27 12000 2 0.349 19
T170306LK2 0.12 80 64 8.6 1U 82B 0.18U 0.18U 14 12000 2 0.354 14
T170306LK3 0.13 76 67 7.6 50B 120B 0.18U 0.18U 21 12000 2.3 0.357 15
T170306LK4 0.12 97 55 16 1U 127B 0.18U 0.18U 18 12000 2.1 0.372 17
Qualifiers
UThecompoundwasanalyzedforbutnotdetected
IThereportedvalueisbetweenthelaboratorymethoddetectionlimitandlaboratoryp
BResultsbaseduponcolonycountsoutsidetheacceptablerange.
FIGURES

Riviera Lake #1
Coffee Pot Bayou
0 1 2 4 6 8
Miles 1 inch = 20,833 feet

USGS The National Map: National Boundaries Dataset, National Elevation Dataset, Geographic Names Information System, National Hydrography Dataset,
Copyright:
National Land Cover Database, National Structures Dataset, and National Transportation Dataset; U.S. Census 2013
Bureau National Geographic
- TIGER/Line; HERE RoadSociety,
Datai-cubed
Water Sampling for Pelican January 2017

Location Map
St. Petersburg, Florida FIGURE 1
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Legend 27 !
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FWC Mortality Reports

29
.
!
Miles 1 inch = 4 miles

USGS The National Map: National Boundaries Dataset, National Elevation Dataset, Geographic Names Information System, National Hydrography Dataset,
Copyright:
National Land Cover Database, National Structures Dataset, and National Transportation Dataset; U.S. Census 2013
Bureau National Geographic
- TIGER/Line; HERE RoadSociety,
Datai-cubed
Pelican Mortality Study FWC Wildlife Mortality Reports March 2017

St. Petersburg, Florida (January 10-30) FIGURE 2

Legend
A
! Water Sample Locations (January)
0 250 500 1,000 1,500 2,000
Feet 1 inch = 1,000 feet
Water Sampling for Pelican March 2017

Sample Locations (January)
St. Petersburg, Florida FIGURE 3
Riviera Bay

T170306LK-3 T170306LK-1
!
( !
(
Riviera Lake #1
#
*
!
(T170306LK-4
T170306LK-2
!
(
th
N or
e
Legend Av
rd
83
#
* Water Quality Location
Sample and Water Quality Location

!
( Riviera Lake #2
0 120 240 480

Feet
Source: Esri, DigitalGlobe, GeoEye, Earthstar Geographics, CNES/Airbus

DS, USDA, USGS, AeroGRID, IGN, and the GIS User Community
Riviera Lake #1 Figure X

March 6, 2017 Sample Locations March 2017
APPENDIX A
Possible
Sources of
Nutrient
Loading
There are a variety of
sources of nutrients and
coliform to lakes:
1 Stormwater runoff from

1
streets, driveways, & lawns 2
2 Pet waste and other wildlife 3
3 Overspray from reclaim
irrigation systems
4 Runoff into lakes from
irrigation and rain events
carries the nutrients into
the lake.
Sample Results: To Tampa Bay

Sampling showed increased 4
nutrients in Riviera Lake #1.
Nutrient-enriched sediment
at bottom of lake
1
Inversion
There was unseasonably warm
weather in Saint Petersburg
this winter. We then had three
days of cold weather and a rain
storm.
1 The cold air and rain caused

an inversion where cold water
fell to the bottom of the lake
and pushed warmer water up to
the surface.
2 The movement of the water

during the inversion disturbed
the nutrient-rich sediments and
naturally low oxygenated water
at the bottom of the lake mixing
throughout the water column.
2
To Tampa Bay
Organism-enriched
sediment at bottom of lake
What caused
the fish kill?
1 An increased number of
micro-organisms mixed with the
nutrients rich water. This
caused an algae bloom that
further consumed oxygen in the
water column.
2 The lack of oxygen

suffocated the fish, causing a
fish kill.
2
To Tampa Bay
1
Organism-enriched
sediment at bottom of lake
Dormant
Our theory on 3
Clostridium 4
botulinum
what caused the 2a Pelican with
pelican deaths? infected fish
The bacteria 1 Clostridium Botulinum

botulinum is naturally present toxin
1
micro-organism in our water 2b
and soil. It thrives in oxygen
depleted environments, so Reanimated
when it senses more oxygen Clostridium
than it likes, 2a it goes botulinum
dormant (and can remain
dormant for a long time). Clostridium
When oxygen levels drop botulinum
and conditions are right, 2b
the Clostridium botulinum will 2b Conditions present in the lake, perfect
reanimate, or wake up, and for generation of Botulinum toxin: Fish with
generate spores 3 that Dissolved Oxygen = 0.32 mg/L botulism
produce the Botulinum toxin Temperature = 21.4 C (71 F)
one of the most potent pH = 6.90
Salinity = 11.33
neurotoxins around. This
toxin can cause botulism in
fish, and if the 4 infected To Tampa Bay
fish are eaten by birds, the
birds will ingest botulism, get
sick and possibly die.
APPENDIX B
FWC Report
Pelican Mortality Update 1/31/2017
Since January 11, 2017, FWC has received 35 reports about dead or ill pelicans across southern Pinellas
County, including eight requests for information. Initial reports were received from Gulfport and Pass-A-
Grille. FWC staff investigated subsequent reports from Riviera Bay Lake #1, Coffee Pot Bayou, and
Bayou Grande and observed dead and sick pelicans.
To date, we have confirmed reports of at least 70 dead or ill brown pelicans and at least one white
pelican. (See map with locations of reports to date below.)
FWC has been working with the City of St. Petersburg, Busch Gardens, Seaside Seabird Sanctuary,
Owls Nest Sanctuary for Wildlife, and other local rehabbers to respond to this event. At least 24
birds have been successfully treated in local wildlife rehabilitation centers and have been released
or are ready to be released.
FWC and partners have collected 23 pelicans for necropsy, although some are not useful due to
the state of decomposition. FWC-FWRI staff at the Wildlife Lab in Gainesville and in St. Petersburg
have necropsied five pelicans; veterinarians at SCWDS (the Southeastern Cooperative Wildlife
Disease Study) in Georgia have necropsied four pelicans; and veterinarians at Busch Gardens have
necropsied at least five pelicans.
Gross necropsies did not yield any remarkable findings. All birds were in good nutritional
condition. Tests for avian influenza in 13 birds and arboviruses in 4 birds have been negative.
Other diagnostic tests (i.e. avian botulism) are still pending.
FWC staff have been monitoring an ongoing bloom of the red tide alga, Karenia brevis, along
Pinellas and Manatee counties (including areas in Lower Tampa Bay). Red tide was not detected in
event response samples collected in Coffee Pot Bayou on 1/13 and just outside the Bayou on 1/18.
Through targeted water sampling conducted on 1/18 in Mid and Lower Tampa Bay, and routine
coastal sampling conducted weekly, FWC has confirmed that bloom levels of Karenia brevis persist
in Lower Tampa Bay. Monitoring for harmful algal blooms is ongoing, and results are updated
biweekly at http://myfwc.com/redtidestatus.
Samples from 15 pelicans were sent to FWC-FWRI staff in St. Petersburg for algal toxin testing to
determine if these mortalities are potentially related to the ongoing red tide. Low levels of
brevetoxin (the red tide toxin) were detected in the gastrointestinal contents of some of the
pelicans, demonstrating some exposure, however tissue samples were negative for the toxin.
Testing is still in progress, and the results to date are inconclusive. Given the low levels measured
and the lack of any other affected species, the deaths may not be related to red tide.
Citizens observing sick or dead birds or other wildlife are encouraged to make an online bird
mortality report or to call FWCs Fish Kill Hotline at 1-800-636-0511.
Locations of sick or dead pelicans reported between January 11 and January 30, 2017.
APPENDIX C
Advanced Environmental Laboratories, Inc
9610 Princess Palm Ave Tampa, FL 33619
Payments: P.O. Box 551580 Jacksonville, FL 32255-1580
Phone: (813)630-9616
Fax: (813)630-4327
February 3, 2017
Kevin Riskowitz
City of St. Petersburg
1635 3rd Ave. N.
Saint Petersburg, FL 33713
RE: Workorder: T1701171 Arcadis St.Pete SW
Dear Kevin Riskowitz:
Enclosed are the analytical results for sample(s) received by the laboratory on Thursday, January 19, 2017. Results reported herein
conform to the most current NELAC standards, where applicable, unless otherwise narrated in the body of the report. The analytical results
for the samples contained in this report were submitted for analysis as outlined by the Chain of Custody and results pertain only to these
samples.
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Enclosures
Report ID: 467256 - 120933 Page 1 of 25
CERTIFICATE OF ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Advanced Environmental Laboratories, Inc.
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
SAMPLE SUMMARY
Workorder: T1701171 Arcadis St.Pete SW
Lab ID Sample ID Matrix Date Collected Date Received
T1701171001 T170119-01 Water 1/19/2017 09:15 1/19/2017 17:15

T1701171002 T170119-02 Water 1/19/2017 09:30 1/19/2017 17:15
T1701171003 T170119-03 Water 1/19/2017 09:45 1/19/2017 17:15
T1701171004 T170119-04 Water 1/19/2017 10:00 1/19/2017 17:15
T1701171005 T170119-05 Water 1/19/2017 10:15 1/19/2017 17:15
T1701171006 T170119-06 Water 1/19/2017 10:35 1/19/2017 17:15
T1701171007 T170119-07 Water 1/19/2017 10:50 1/19/2017 17:15
T1701171008 T170119LK-01 Water 1/19/2017 13:50 1/19/2017 17:15
T1701171009 T170119LK-02 Water 1/19/2017 14:20 1/19/2017 17:15
T1701171010 T170119LK-03 Water 1/19/2017 14:45 1/19/2017 17:15
Report ID: 467256 - 120933 Page 2 of 25
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ANALYTICAL RESULTS
Lab ID: T1701171001 Date Received: 01/19/17 17:15 Matrix: Water

Sample ID: T170119-01 Date Collected: 01/19/17 09:15
RegLmt
Sample Description: Location:
Adjusted Adjusted
Parameters Results Qual Units DF PQL MDL Analyzed Lab
Microbiology
Analysis Desc: Total Analytical Method: SM 9222 B (MF)
Coliform,SM9222B,Water
Coliform Total 1320 B #/100 mL 9 9 9 1/20/2017 09:00 T
Analysis Desc: Fecal Coliform Analytical Method: SM 9222D

MF,SM9222D,Water
Coliform Fecal 918 B #/100 mL 9 9 9 1/19/2017 18:10 T
WET CHEMISTRY
Analysis Desc: Ammonia,E350.1,Water Analytical Method: EPA 350.1
Ammonia (N) 0.02 U mg/L 1 0.10 0.02 1/25/2017 14:16 T
Analysis Desc: TKN,E351.2,Water Preparation Method: Copper Sulfate Digestion

Analytical Method: EPA 351.2
Total Kjeldahl Nitrogen 0.46 mg/L 1 0.20 0.075 1/24/2017 12:06 T
Analysis Desc: Total Preparation Method: EPA 365.3

Phosphorus,E365.3,Analysis
Total Phosphorus (as P) 0.109 mg/L 1 0.01 0.005 2/2/2017 09:05 G
Analysis Desc: Chlorophylls, Analytical Method: SM 10200 H

SM10200H,Water
Chlorophyll A 9.1 mg/m3 1 1.0 1.0 1/30/2017 13:30 G
Chlorophyll B 1.0 U mg/m3 1 1.0 1.0 1/30/2017 13:30 G
Chlorophyll C 1.0 U mg/m3 1 1.0 1.0 1/30/2017 13:30 G
Pheophytin A 1.0 U mg/m3 1 1.0 1.0 1/30/2017 13:30 G
Analysis Desc: Tot Dissolved Analytical Method: SM 2540 C

Solids,SM2540C
Total Dissolved Solids 26000 mg/L 1.25 12 12 1/25/2017 14:58 T
Analysis Desc: TSS,SM2540D,Water Analytical Method: SM 2540D

Total Suspended Solids 15 mg/L 1 1.0 1.0 1/25/2017 07:44 T
Analysis Desc: Nitrate,Nitrite Analytical Method: SM 4500NO3-F

SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:26 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:26 T
Report ID: 467256 - 120933 Page 3 of 25
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ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
Coliform Fecal 1 U #/100 mL 1 1 1 1/19/2017 18:10 T
WET CHEMISTRY
Ammonia (N) 0.02 U mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water
Pheophytin A 1.6 mg/m3 1 1.0 1.0 1/30/2017 13:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:27 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:27 T
Report ID: 467256 - 120933 Page 4 of 25
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Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.02 U mg/L 1 0.10 0.02 1/25/2017 14:16 T


Total Phosphorus (as P) 0.008 I mg/L 1 0.01 0.005 2/2/2017 09:05 G

SM10200H,Water

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:28 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:28 T
Report ID: 467256 - 120933 Page 5 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.02 U mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water
Chlorophyll C 1.5 mg/m3 1 1.0 1.0 1/30/2017 13:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:29 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:29 T
Report ID: 467256 - 120933 Page 6 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.02 U mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water
Chlorophyll A 10 mg/m3 1 1.0 1.0 1/30/2017 13:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:30 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:30 T
Report ID: 467256 - 120933 Page 7 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.02 U mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water
Chlorophyll B 1.2 mg/m3 1 1.0 1.0 1/30/2017 13:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:31 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:31 T
Report ID: 467256 - 120933 Page 8 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology
Coliform Total 1 U #/100 mL 1 1 1 1/20/2017 09:00 T

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.02 U mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:32 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:32 T
Report ID: 467256 - 120933 Page 9 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

Sample ID: T170119LK-01 Date Collected: 01/19/17 13:50
RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.71 mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water
Chlorophyll B 12 mg/m3 1 1.0 1.0 1/30/2017 13:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:39 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:39 T
Report ID: 467256 - 120933 Page 10 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.30 mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:40 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:40 T
Report ID: 467256 - 120933 Page 11 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.10 mg/L 1 0.10 0.02 1/25/2017 14:16 T



SM10200H,Water
Chlorophyll C 11 mg/m3 1 1.0 1.0 1/30/2017 13:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:41 T
Nitrite 0.18 U mg/L 1 0.20 0.18 1/20/2017 15:41 T
Report ID: 467256 - 120933 Page 12 of 25
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ANALYTICAL RESULTS QUALIFIERS
PARAMETER QUALIFIERS
U The compound was analyzed for but not detected.
I The reported value is between the laboratory method detection limit and the laboratory practical quantitation limit.
B Results based upon colony counts outside the acceptable range.
LAB QUALIFIERS
G DOH Certification #E82001(AEL-G)(FL NELAC Certification)
T DOH Certification #E84589(AEL-T)(FL NELAC Certification)
Report ID: 467256 - 120933 Page 13 of 25
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Fax: (813)630-4327
QUALITY CONTROL DATA
QC Batch: WCAt/6724 Analysis Method: SM 4500NO3-F

QC Batch Method: SM 4500NO3-F Prepared:
Associated Lab Samples: T1701171001, T1701171002, T1701171003, T1701171004, T1701171005, T1701171006, T1701171007, T1701171008,
METHOD BLANK: 2252759
Blank Reporting
Parameter Units Result Limit Qualifiers
WET CHEMISTRY
Nitrate mg/L 0.18 0.18 U
Nitrite mg/L 0.18 0.18 U
LABORATORY CONTROL SAMPLE: 2252760
Spike LCS LCS % Rec

Parameter Units Conc. Result % Rec Limits Qualifiers
WET CHEMISTRY
Nitrate mg/L 1 1.1 110 90-110
Nitrite mg/L 1 0.99 99 90-110
MATRIX SPIKE & MATRIX SPIKE DUPLICATE: 2253106 2253107 Original: T1701205001
Original Spike MS MSD MS MSD % Rec Max

Parameter Units Result Conc. Result Result % Rec % Rec Limit RPD RPD Qualifiers
WET CHEMISTRY
Nitrate mg/L 0.59 1 1.6 1.6 98 102 90-110 3 10
Nitrite mg/L 0 1 0.98 0.98 98 98 90-110 0 10
QC Batch: WCAt/6743 Analysis Method: EPA 351.2

QC Batch Method: Copper Sulfate Digestion Prepared: 01/23/2017 18:00
Blank Reporting
WET CHEMISTRY
Total Kjeldahl Nitrogen mg/L 0.075 0.075 U
Report ID: 467256 - 120933 Page 14 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Blank Reporting
WET CHEMISTRY
Spike LCS LCS % Rec

WET CHEMISTRY
Total Kjeldahl Nitrogen mg/L 1 1.1 106 90-110
Spike LCS LCS % Rec

WET CHEMISTRY

WET CHEMISTRY
Total Kjeldahl Nitrogen mg/L 0.3 1 1.4 1.4 110 108 90-110 1 20

WET CHEMISTRY
Total Kjeldahl Nitrogen mg/L 1.0 1.0 3 20

Report ID: 467256 - 120933 Page 15 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Blank Reporting
WET CHEMISTRY
Blank Reporting
WET CHEMISTRY
Spike LCS LCS % Rec

WET CHEMISTRY
Spike LCS LCS % Rec

WET CHEMISTRY

WET CHEMISTRY

WET CHEMISTRY
Total Kjeldahl Nitrogen mg/L 1.0 1.0 3 20
Report ID: 467256 - 120933 Page 16 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
QC Batch: MICt/2397 Analysis Method: SM 9222D

QC Batch Method: SM 9222D Prepared:
Blank Reporting
Microbiology
Coliform Fecal #/100 mL 1 1 U
QC Batch: MICt/2407 Analysis Method: SM 9222 B (MF)

QC Batch Method: SM 9222 B (MF) Prepared:
Blank Reporting
Microbiology
Coliform Total #/100 mL 1 1 U
QC Batch: WCAt/6748 Analysis Method: SM 2540D

Associated Lab Samples: T1701171001, T1701171002, T1701171003, T1701171004
Blank Reporting
WET CHEMISTRY
Total Suspended Solids mg/L 1.0 1.0 U
Spike LCS LCS % Rec

WET CHEMISTRY
Total Suspended Solids mg/L 200 210 103 75-125
Report ID: 467256 - 120933 Page 17 of 25
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Phone: (813)630-9616
Fax: (813)630-4327
SAMPLE DUPLICATE: 2254914 Original: T1701147003
Original DUP Max

Parameter Units Result Result RPD RPD Qualifiers
WET CHEMISTRY
Total Suspended Solids mg/L 1.0U 1.0 0 10
Associated Lab Samples: T1701171005, T1701171006, T1701171007, T1701171008, T1701171009, T1701171010
Blank Reporting
WET CHEMISTRY
Spike LCS LCS % Rec

WET CHEMISTRY
Original DUP Max

WET CHEMISTRY
Total Suspended Solids mg/L 15 14 6 10
Original DUP Max

WET CHEMISTRY
Total Suspended Solids mg/L 1.2 1.2 0 10
Report ID: 467256 - 120933 Page 18 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
QC Batch: WCAt/6769 Analysis Method: SM 2540 C

QC Batch Method: SM 2540 C Prepared:
Blank Reporting
WET CHEMISTRY
Total Dissolved Solids mg/L 10 10 U
Spike LCS LCS % Rec

WET CHEMISTRY
Total Dissolved Solids mg/L 660 620 94 75-125
Original DUP Max

WET CHEMISTRY
Total Dissolved Solids mg/L 390 380 3 10
Original DUP Max

WET CHEMISTRY
QC Batch Method: EPA 350.1 Prepared:
Blank Reporting
WET CHEMISTRY
Ammonia (N) mg/L 0.02 0.02 U
Report ID: 467256 - 120933 Page 19 of 25
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Spike LCS LCS % Rec

WET CHEMISTRY
Ammonia (N) mg/L 0.5 0.55 109 90-110

WET CHEMISTRY
Ammonia (N) mg/L 0 1 1.0 1.00 101 100 90-110 2 10

WET CHEMISTRY
Ammonia (N) mg/L 0 1 0.95 0.91 95 91 90-110 4 10
QC Batch: WCAg/4089 Analysis Method: SM 10200 H

QC Batch Method: SM 10200 H Prepared:
Blank Reporting
WET CHEMISTRY
Chlorophyll A mg/m3 1.0 1.0 U
Chlorophyll B mg/m3 1.0 1.0 U
Chlorophyll C mg/m3 1.0 1.0 U
Pheophytin A mg/m3 1.0 1.0 U
Report ID: 467256 - 120933 Page 20 of 25
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Original DUP Max

WET CHEMISTRY
Chlorophyll A mg/m3 9.1 9.1 0 20
Chlorophyll B mg/m3 1.0U 1.0 0
Chlorophyll C mg/m3 1.0U 1.0 0
Pheophytin A mg/m3 1.0U 1.0 0
QC Batch: WCAg/4112 Analysis Method: EPA 365.3
QC Batch Method: EPA 365.3 Prepared: 02/01/2017 15:40
Blank Reporting
WET CHEMISTRY
Total Phosphorus (as P) mg/L 0.005 0.005 U
Spike LCS LCS % Rec

WET CHEMISTRY
Total Phosphorus (as P) mg/L 0.1 0.104 104 80-120

WET CHEMISTRY
Total Phosphorus (as P) mg/L 0.109 0.25 0.353 0.348 98 96 80-120 1 20
Report ID: 467256 - 120933 Page 21 of 25
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
QUALITY CONTROL DATA CROSS REFERENCE TABLE
Analysis
Lab ID Sample ID Prep Method Prep Batch Analysis Method Batch
T1701171001 T170119-01 SM 4500NO3-F WCAt/6724

T1701171002 T170119-02 SM 4500NO3-F WCAt/6724
T1701171003 T170119-03 SM 4500NO3-F WCAt/6724
T1701171004 T170119-04 SM 4500NO3-F WCAt/6724
T1701171005 T170119-05 SM 4500NO3-F WCAt/6724
T1701171006 T170119-06 SM 4500NO3-F WCAt/6724
T1701171007 T170119-07 SM 4500NO3-F WCAt/6724
T1701171008 T170119LK-01 SM 4500NO3-F WCAt/6724
T1701171009 T170119LK-02 SM 4500NO3-F WCAt/6724
T1701171010 T170119LK-03 SM 4500NO3-F WCAt/6724
T1701171001 T170119-01 Copper Sulfate Digestion WCAt/6743 EPA 351.2 WCAt/6767

T1701171008 T170119LK-01 Copper Sulfate Digestion WCAt/6743 EPA 351.2 WCAt/6767
T1701171001 T170119-01 SM 9222D MICt/2397

T1701171002 T170119-02 SM 9222D MICt/2397
T1701171003 T170119-03 SM 9222D MICt/2397
T1701171004 T170119-04 SM 9222D MICt/2397
T1701171005 T170119-05 SM 9222D MICt/2397
T1701171006 T170119-06 SM 9222D MICt/2397
T1701171007 T170119-07 SM 9222D MICt/2397
T1701171008 T170119LK-01 SM 9222D MICt/2397
T1701171009 T170119LK-02 SM 9222D MICt/2397
T1701171010 T170119LK-03 SM 9222D MICt/2397
T1701171001 T170119-01 SM 9222 B (MF) MICt/2407
Report ID: 467256 - 120933 Page 22 of 25
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Phone: (813)630-9616
Fax: (813)630-4327
Analysis
T1701171002 T170119-02 SM 9222 B (MF) MICt/2407

T1701171003 T170119-03 SM 9222 B (MF) MICt/2407
T1701171004 T170119-04 SM 9222 B (MF) MICt/2407
T1701171005 T170119-05 SM 9222 B (MF) MICt/2407
T1701171006 T170119-06 SM 9222 B (MF) MICt/2407
T1701171007 T170119-07 SM 9222 B (MF) MICt/2407
T1701171008 T170119LK-01 SM 9222 B (MF) MICt/2407
T1701171009 T170119LK-02 SM 9222 B (MF) MICt/2407
T1701171010 T170119LK-03 SM 9222 B (MF) MICt/2407
T1701171001 T170119-01 SM 2540D WCAt/6748

T1701171002 T170119-02 SM 2540D WCAt/6748
T1701171003 T170119-03 SM 2540D WCAt/6748
T1701171004 T170119-04 SM 2540D WCAt/6748
T1701171005 T170119-05 SM 2540D WCAt/6749

T1701171006 T170119-06 SM 2540D WCAt/6749
T1701171007 T170119-07 SM 2540D WCAt/6749
T1701171008 T170119LK-01 SM 2540D WCAt/6749
T1701171009 T170119LK-02 SM 2540D WCAt/6749
T1701171010 T170119LK-03 SM 2540D WCAt/6749
T1701171001 T170119-01 SM 2540 C WCAt/6769

T1701171002 T170119-02 SM 2540 C WCAt/6769
T1701171003 T170119-03 SM 2540 C WCAt/6769
T1701171004 T170119-04 SM 2540 C WCAt/6769
T1701171005 T170119-05 SM 2540 C WCAt/6769
T1701171006 T170119-06 SM 2540 C WCAt/6769
T1701171007 T170119-07 SM 2540 C WCAt/6769
T1701171008 T170119LK-01 SM 2540 C WCAt/6769
T1701171009 T170119LK-02 SM 2540 C WCAt/6769
T1701171010 T170119LK-03 SM 2540 C WCAt/6769
Report ID: 467256 - 120933 Page 23 of 25
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Phone: (813)630-9616
Fax: (813)630-4327
Analysis
T1701171001 T170119-01 EPA 350.1 WCAt/6771

T1701171002 T170119-02 EPA 350.1 WCAt/6771
T1701171003 T170119-03 EPA 350.1 WCAt/6771
T1701171004 T170119-04 EPA 350.1 WCAt/6771
T1701171005 T170119-05 EPA 350.1 WCAt/6771
T1701171006 T170119-06 EPA 350.1 WCAt/6771
T1701171007 T170119-07 EPA 350.1 WCAt/6771
T1701171008 T170119LK-01 EPA 350.1 WCAt/6771
T1701171009 T170119LK-02 EPA 350.1 WCAt/6771
T1701171010 T170119LK-03 EPA 350.1 WCAt/6771
T1701171001 T170119-01 SM 10200 H WCAg/4089

T1701171002 T170119-02 SM 10200 H WCAg/4089
T1701171003 T170119-03 SM 10200 H WCAg/4089
T1701171004 T170119-04 SM 10200 H WCAg/4089
T1701171005 T170119-05 SM 10200 H WCAg/4089
T1701171006 T170119-06 SM 10200 H WCAg/4089
T1701171007 T170119-07 SM 10200 H WCAg/4089
T1701171008 T170119LK-01 SM 10200 H WCAg/4089
T1701171009 T170119LK-02 SM 10200 H WCAg/4089
T1701171010 T170119LK-03 SM 10200 H WCAg/4089
T1701171001 T170119-01 EPA 365.3 WCAg/4112 EPA 365.3 WCAg/4113

T1701171008 T170119LK-01 EPA 365.3 WCAg/4112 EPA 365.3 WCAg/4113
Report ID: 467256 - 120933 Page 24 of 25
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Page 25 of 25
APPENDIX D
4985 SW 74th Court, Miami, FL 33155 USA
Tel: (1) 786-220-0379, Fax: (1) 786-513-2733, Email: info@sourcemolecular.com
Preliminary Interpretation of Bird Fecal IDTM Quantification Results

Detection and quantification of Bird-associated fecal indicator bacteria by real-time quantitative
Polymerase Chain Reaction (qPCR)
Submitter: Arcadis
Date Received: January 20, 2017
Report Generated: February 27, 2017
Approximate Contribution
SM # Client # of Bird Fecal Pollution in Comment
Water Sample
SM-7B07007 T170119-01 Low Concentration Low levels of Bird fecal biomarker

SM-7B07008 T170119-06 Low Concentration Low levels of Bird fecal biomarker
SM-7B07009 T170119LK-01 Low Concentration Low levels of Bird fecal biomarker
Limitation of Damages Repayment of Service Price

It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its
agents or representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis
price paid by him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source
Molecular Corp. provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited
in this report may be used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is
not limited to internal validation studies and peer-reviewed research publications. Anonymity of the sample(s), including the exact geographic
location will be maintained by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of
origin for research purposes. The client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not
wish for their submitted sample(s) to be used for any type of future research.
Tel: (1) 786-220-0379, Fax: (1) 786-513-2733, Email:
Bird Fecal IDTM Quantification

Detection and quantification of Bird-associated fecal indicator bacteria by real-time
quantitative Polymerase Chain Reaction (qPCR)
Submitter: Arcadis
Bird Specific Marker DNA Analytical

SM # Client # Analysis Requested
Quantified* Results
SM-7B07007 T170119-01 Bird Fecal ID <LOQ Present

SM-7B07008 T170119-06 Bird Fecal ID <LOQ Present
SM-7B07009 T170119LK-01 Bird Fecal ID <LOQ Present
*Numbers reported as copy numbers per 100 mL of water
<LOQ: Detected below level of quantification
Laboratory Comments
Submitter: Arcadis
Negative Results
In sample(s) classified as negative, the bird-associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis. It is important to note that a
negative result does not mean that the sample does not definitely have bird fecal contamination. Only repeated
sampling (both during wet and dry sampling events) will enable you to draw more definitive conclusions as to the
contributor(s) of fecal pollution.
Positive Results
In sample(s) classified as positive, the bird-associated fecal gene biomarker(s) was detected in both test replicates
suggesting that bird fecal contamination is present in the water sample(s). The biomarker(s) serve as an indicator of
the targeted fecal pollution, but the presence of the biomarker does not signify conclusively the presence of that form
of fecal pollution. Only repeated sampling (both during wet and dry sampling events) will enable you to draw more
definitive conclusions as to the contributor(s) of fecal pollution.
<LOQ Results
In sample(s) classified as <LOQ, the bird-associated fecal biomarker was detected in both test replicates but in
quantities below the limit of quantification. This result indicates that fecal indicators associated with bird were present
in the sample(s) but in low concentrations.
Bird Fecal Reference Samples

The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the bird-associated fecal genetic in the geographic region of interest. A
more precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
Quantitative results are reported along with interpretations. Interpretations are given as "negative", trace, "low
concentration", "moderate concentration", or "high concentration" based on the concentration of the genetic markers
found in the water samples.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination. A list of available
tests can be found at sourcemolecular.com/tests
DNA Analytical Method Explanation

Each submitted water sample was filtered through 0.45 micron membrane filters. Each filter was placed in a separate,
sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample was homogenized for 1min
and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's protocol.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul containing sample extract, forward primer,
reverse primer and an optimized buffer. All assays were run in duplicate. Absolute quantification was achieved by
extrapolating target gene copy numbers from a standard curve generated from serial dilutions of known gene copy
numbers.
For quality control purposes, a positive control consisting of bird fecal DNA or plasmid containing the target and a negative
control consisting of PCR-grade water, were run alongside the sample(s) to ensure a properly functioning reaction and to
reveal any false negatives or false positives. The accumulation of PCR product was detected and graphed in an
amplification plot. If the target gene biomarker was absent in the sample, this accumulation was not detected and the
sample was considered negative. If accumulation of PCR product was detected, the sample was considered positive.
Theory Explanation of Bird Fecal IDTM Quantification
The genus Helicobacter is a group of gram-negative, microaerophilic bacteria that were initially classified
under the Campylobacter genus prior to 1989. Since then, they have been reclassified into the genus
Helicobacter after 16S rRNA sequencing differentiated them from other Campylobacter species. This group
of bacteria typically have a spiral, curved or fusiform morphology with multiple flagella allowing them to
rapidly maneuver in the intestinal mucous lining of their hosts. Helicobacter species colonize the
gastrointestinal tract of mammals and birds and are shed in feces. There are approximately 20 strains of
Helicobacter1. Certain strains, such as Helicobacter pylori, are pathogenic to humans causing chronic
gastritis, peptic ulcers and stomach cancer.
The Bird Fecal Quantification ID service is designed around the principle that certain DNA sequences
contained within strains of the Helicobacter genus are specific to wild birds. These Helicobacter sequences
can be used as indicators of bird fecal contamination. Several species have been isolated from specific
animal hosts such as H. fennelliae from humans, H. hepaticus from mice and H. felis from cats and dogs.1
The Bird Fecal Quantification ID service targets a bird-associated gene biomarker in Helicobacter
pametensis.2 The biomarker is present at different degrees in the feces of various birds including but not
limited to gull, goose, chicken, pigeon and duck.
One of the advantages of the Bird Fecal Quantification ID service is that the entire population of
Helicobacter of the selected portion of the water sample is screened. As such, this method avoids the
randomness effect of selecting isolates off a petri dish.
Accuracy of the results is possible because the method uses qPCR DNA technology. qPCR
simulataneously confirms and quantifies the bird-associated gene biomarker. This is accomplished with
small pieces of DNA called primers that are complementary and specific to the genome to be detected. This
qPCR technology avoids the cumbersome process of distinguishing DNA bands on a gel electrophoresis
apparatus.
Through a heating process called thermal cycling, the double stranded DNA is denatured and inserted with
complementary primers to create exact copies of the DNA fragment desired. This process is repeated
rapidly many times ensuring an exponential progression in the number of copied DNA. If the primers are
successful in finding a site on the DNA fragment that is specific to the genome to be studied, then billions of
copies of the DNA fragment will be available and detected in real-time. The accumulation of DNA product is
plotted as an amplification curve. The absence of an amplification curve indicates that the bird-associated
Helicobacter gene biomarker is not present.
References
1 Goldman, E. and Green, L. H. (2009). Practical Handbook of Microbiology (2nd ed) . Boca Raton, FL: CRC Press.
2 Seymour, C., Lewis, R.G., Kim, M., Gagnon, D.F., Fox, J.G., Dewhirst, F.E., and Paster, B.J. Isolation of Helicobacter Strains
from Wild Bird and Swine Feces. Appl. Environ. Microbiol. (1994) 60:3, 1025-1028.
Preliminary Interpretation of Dog Quantification IDTM Results

Detection and quantification of the fecal Dog gene biomarker for Dog fecal contamination by real-time
quantitative Polymerase Chain Reaction (qPCR) DNA analytical technology
Submitter: Arcadis
Approximate Contribution
SM # Client # of Dog Fecal Pollution in Comment
Water Sample
SM-7B07004 T170119-01 Not Detected Dog fecal biomarker not detected

SM-7B07005 T170119-06 Not Detected Dog fecal biomarker not detected
SM-7B07006 T170119LK-01 Not Detected Dog fecal biomarker not detected

Molecular Corp. provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited
in this report may be used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is
not limited to internal validation studies and peer-reviewed research publications. Anonymity of the sample(s), including the exact geographic
Tel: (1) 786-220-0379, Fax: (1) 786-513-2733, Email:
Dog Bacteroidetes Quantification IDTM

Detection and quantification of the fecal Dog gene biomarker for Dog fecal
contamination by real-time quantitative Polymerase Chain Reaction (qPCR) DNA
Submitter: Arcadis
Marker Quantified DNA Analytical

SM # Client # Analysis Requested
(copies/100 ml) Results
SM-7B07004 T170119-01 Dog Bacteroidetes ID ND Absent

SM-7B07005 T170119-06 Dog Bacteroidetes ID ND Absent
SM-7B07006 T170119LK-01 Dog Bacteroidetes ID ND Absent
ND: Not Detected
Laboratory Comments
Submitter: Arcadis
Negative Results
In sample(s) classified as negative, the dog-associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis. It is important to note that a
negative result does not mean that the sample does not definitely have dog fecal contamination. Only repeated
sampling (both during wet and dry sampling events) will enable you to draw more definitive conclusions as to the
contributor(s) of fecal pollution.
Positive Results
In sample(s) classified as positive, the dog-associated fecal gene biomarker(s) was detected in both test replicates
suggesting that dog fecal contamination is present in the water sample(s). The biomarker(s) serve as an indicator of
the targeted fecal pollution, but the presence of the biomarker does not signify conclusively the presence of that form of
fecal pollution. Only repeated sampling (both during wet and dry sampling events) will enable you to draw more
definitive conclusions as to the contributor(s) of fecal pollution.
<LOQ Results
In sample(s) classified as <LOQ, the dog-associated fecal biomarker was detected in both test replicates but in
quantities below the limit of quantification. This result indicates that fecal indicators associated with dog were present in
the sample(s) but in low concentrations.
Dog Fecal Reference Samples

The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the dog-associated fecal genetic marker in the geographic region of
interest. A more precise interpretation would be available to the client with the submittal of such baseline samples.
Additional Testing
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination. A list of available
tests can be found at sourcemolecular.com/tests

Each submitted water sample was filtered through 0.45 micron membrane filters. Each filter was placed in a separate,
sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample was homogenized for 1min
and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's protocol.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real -time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul containing sample extract, forward primer,
reverse primer, probe and an optimized buffer. The following thermal cycling parameters were used: 95 C for 10 min and
40 cycles of 95C for 15 s and 60C for 1 min. All assays were run in duplicate. Absolute quantification was achieved by
extrapolating target gene copy numbers from a standard curve generated from serial dilutions of known gene copy
numbers.
For quality control purposes, a positive control consisting of Dog fecal DNA and a negative control consisting of PCR -
grade water, were run alongside the sample(s) to ensure a properly functioning reaction and reveal any false negatives or
false positives. The accumulation of PCR product is detected and graphed in an amplification plot. If the fecal indicator
organism is absent in the sample, this accumulation is not detected and the sample is considered negative. If
accumulation of PCR product is detected, the sample is considered positive.
Theory Explanation of Dog Bacteroidetes Quantification IDTM
The phylum Bacteroidetes is composed of three large groups of bacteria with the best-known category
being Bacteroidaceae. This family of gram-negative bacteria is found primarily in the intestinal tracts and
mucous membranes of warm-blooded animals and is sometimes considered pathogenic.
Comprising Bacteroidaceae are the genus Bacteroides and Prevotella. The latter genus was originally
classified within the former (i.e. Bacteroides), but since the 1990s it has been classified in a separate genus
because of new chemical and biochemical findings. Bacteroides and Prevotella are gram-negative,
anaerobic, rod-shaped bacteria that inhabitant of the oral, respiratory, intestinal, and urogenital cavities of
humans, animals, and insects. They are sometimes pathogenic.
Fecal Bacteroidetes are considered for several reasons an interesting alternative to more traditional
indicator organisms such as E. coli and Enterococci.1 Since they are strict anaerobes, they are indicative of
recent fecal contamination when found in water systems. This is a particularly strong reference point when
trying to determine recent outbreaks in fecal pollution. They are also more abundant in feces of warm-
blooded animals than E. coli and Enterococci. Furthermore, these latter two organisms are facultative
anaerobes and as such they can be problematic for monitoring purposes since it has been shown that they
are able to proliferate in soil, sand and sediments.
The Dog Bacteroidetes IDTM service is designed around the principle that fecal Bacteroidetes are found in
large quantities in feces of warm-blooded animals.2,3,4,5,6 Furthermore, certain categories of Bacteroidetes
have been shown to be predominately detected in dog. Within these Bacteroidetes, certain strains of the
Bacteroides and Prevotella genus have been found in dog.2,3,5,6 As such, these bacterial strains can be used
as indicators of dog fecal contamination.
One of the advantages of the Dog Bacteroidetes IDTM service is that the entire water is sampled and filtered
for fecal Bacteroidetes. As such, this method avoids the randomness effect of culturing and selecting
bacterial isolates off a petri dish. This is a particular advantage for highly contaminated water systems with
potential multiple sources of fecal contamination.
Accuracy of the results is possible because the method uses PCR DNA technology. PCR allows quantities
of DNA to be amplified into large number of small copies of DNA sequences. This is accomplished with
small pieces of DNA called primers that are complementary and specific to the genomes to be detected.
Through a heating process called thermal cycling, the double stranded DNA is denatured and inserted with
complementary primers to create exact copies of the DNA fragment desired. This process is repeated
rapidly many times ensuring an exponential progression in the number of copied DNA. If the primers are
successful in finding a site on the DNA fragment that is specific to the genome to be studied, then billions of
copies of the DNA fragment will be available and detected in real-time. The accumulation of DNA product is
plotted as an amplification curve. The absence of an amplification curve would indicate that the dog
Bacteroidetes gene biomarker is not present.
References
1 Scott, Troy M., Rose, Joan B., Jenkins, Tracie M., Farrah, Samuel R., Lukasik, Jerzy Microbial Source Tracking: Current Methodology and
Future Directions. Appl. Environ. Microbiol. (2002) 68: 5796-5803.

2 Bernhard, A.E., and K.G. Field (2000a). Identification of nonpoint sources of fecal pollution in coastal waters by using host-specific 16S
ribosomal DNA genetic markers from fecal anaerobes. Applied and Environmental Microbiology, 66: 1,587-1,594.
3 Bernhard, A.E., and K.G. Field (2000b). A PCR assay to discriminate human and ruminant feces on the basis of host differences in
Bacteroides-Prevotella genes encoding 16S rRNA. Applied and Environmental Microbiology, 66: 4,571-4,574.
4 Kreader, C.A. (1995). Design and evaluation of Bacteroides DNA probes for the specific detection of human fecal pollution. Applied and
Environmental Microbiology, 61: 1,171-1,179.

5 Fogarty, Lisa R., Voytek, Mary A.Comparison of Bacteroides-Prevotella 16S rRNA Genetic Markers for Fecal Samples from Different
Animal Species Appl. Environ. Microbiol. 2005 71: 5999-6007.

6 Dick, Linda K., Bernhard, Anne E., Brodeur, Timothy J., Santo Domingo, Jorge W., Simpson, Joyce M., Walters, Sarah P., Field, Katharine G. Host
Preliminary Interpretation of Human Fecal Pollution IDTM Results

Detection and quantification of the fecal Human gene biomarker for Human fecal contamination by real-time
quantitative Polymerase Chain Reaction (qPCR) DNA analytical technology
Submitter: Arcadis
Approximate
Contribution of Human
SM # Client # Comment
Fecal Pollution in Water
Sample
SM-7A20004 T170119-01 Low Concentration Low levels of Human fecal biomarker
SM-7A20009 T170119-06 Not Detected Human fecal biomarker not detected
SM-7A20011 T170119LK-01 Low Concentration Low levels of Human fecal biomarker

Molecular Corp. provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in
this report may be used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not
limited to internal validation studies and peer-reviewed research publications. Anonymity of the sample(s), including the exact geographic
Human Fecal Pollution IDTM Quantification

Detection and quantification of the fecal Human gene biomarker for Human fecal contamination by
real-time quantitative Polymerase Chain Reaction (qPCR) DNA analytical technology
Submitter: Arcadis
Marker Quantified DNA Analytical

SM # Client # Analysis Requested Target
(copies/100 ml) Results
SM-7A20004 T170119-01 Human Bacteroidetes ID 1 Dorei ND Absent

SM-7A20009 T170119-06 Human Bacteroidetes ID 1 Dorei ND Absent
SM-7A20011 T170119LK-01 Human Bacteroidetes ID 1 Dorei <LOQ Present
SM-7B07001 T170119-01 Human Bacteroidetes ID 2 EPA <LOQ Present
SM-7B07002 T170119-06 Human Bacteroidetes ID 2 EPA ND Absent
SM-7B07003 T170119LK-01 Human Bacteroidetes ID 2 EPA ND Absent
ND - Non-detect
<LOQ: Detected below level of quantification
Laboratory Comments
Submitter: Arcadis
Negative Results
In sample(s) classified as negative, the human-associated Bacteroidetes gene biomarker(s) was either not detected
in test replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one
replicate was detected at a cycle threshold less than 35 and the other was not after repeated analysis. It is important
to note that a negative result does not mean that the sample does not definitely have human fecal contamination.
Only repeated sampling (both during wet and dry sampling events) will enable you to draw more definitive
conclusions as to the contributor(s) of fecal pollution.
In order to strengthen the result, a negative sample should be analyzed further for human fecal contamination with
other DNA analytical tests. A list of human fecal ID tests can be found at www.sourcemolecular.com/human.
Positive Results
In sample(s) classified as positive, the human-associated Bacteroidetes gene biomarker(s) was detected in both test
replicates suggesting that human fecal contamination is present in the water sample(s). The biomarker(s) serve as an
indicator of the targeted fecal pollution, but the presence of the biomarker does not signify conclusively the presence
of that form of fecal pollution. Only repeated sampling (both during wet and dry sampling events) will enable you to
draw more definitive conclusions as to the contributor(s) of fecal pollution.
<LOQ Results
In sample(s) classified as <LOQ, the human-associated Bacteroidetes biomarker was detected in both test replicates
but in quantities below the limit of quantification. This result indicates that fecal indicators associated with human
were present in the sample(s) but in low concentrations.
Human Fecal Reference Samples

The client is encouraged to submit samples from the surrounding wastewater facilities and/or septic systems in order
to gain a better understanding of the concentration of the human-associated fecal Bacteroidetes genetic marker as
well as the concentration of the general fecal Bacteroidetes genetic marker in the geographic region of interest. A
more precise interpretation would be available to the client with the submittal of such baseline samples.
Additional Testing
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination. A list of
available tests can be found at www.sourcemolecular.com/tests

All reagents, chemicals and apparatuses were verified and inspected beforehand to ensure that no false negatives or positives
could be generated. In that regard, positive and negative controls were run to attest the integrity of the analysis. All inspections and
controls tested negative for possible extraneous contaminates, including PCR inhibitors.
Each submitted water sample was filtered through 0.45 micron membrane filters. Each filter was placed in a separate, sterile 2ml
disposable tube containing a unique mix of beads and lysis buffer. The sample was homogenized for 1min and the DNA extracted
using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's protocol.
Amplifications were run on an Applied Biosystems StepOnePlus real-time thermal cycler (Applied Biosystems, Foster City, CA) in a
final reaction volume of 20ul containing sample extract, forward primer, reverse primer, probe and an optimized buffer. The
following thermal cycling parameters were used: 50C for 2 min, 95C for 10 min and 40 cycles of 95C for 15 s and 60C for 1 min.
All assays were run in duplicate. Absolute quantification was achieved by extrapolating genome copy numbers from standard
curves generated from serial dilutions of Human specific and generic genomic DNA.
For quality control purposes, a positive control consisting of appropriate genomic DNA and a negative control consisting of PCR-
grade water were run alongside the sample(s) to ensure a properly functioning reaction and reveal any false negatives or false
positives.
Human Bacteroidetes IDTM Species: B. dorei
The Human Bacteroidetes IDTM Species: B. dorei service targets the species Bacteroides dorei. B. dorei
is an anaerobe that is frequently shed from the gastrointestinal tract and isolated from human feces
worldwide. It is a newly discovered species that is widely distributed in the USA.1,2 The human-associated
marker DNA sequence is located on the 16S rRNA gene of B. dorei.3 The marker is the microbial source
tracking (MST) marker of choice for detecting human fecal pollution due to its exceptional sensitivity and
specificity. Internal validations have been conducted on hundreds of sewage, septage, human and animal
host fecal samples collected from throughout the U.S and archived in the Source Molecular fecal bank. The
marker has also been evaluated in both inland and coastal waters. A recent, comprehensive, multi-
laboratory MST method evaluation study, exploring the performance of current MST methods, concluded
the B. dorei qPCR assay to be the top performing human-associated assay amongst those tested. The
success and consistency of this marker in numerous studies around the world1,3,4 makes the Human
Bacteroidetes IDTM Species: B. dorei service the primary service for identifying human fecal pollution at
Source Molecular.
Fecal Bacteroidetes are considered for several reasons an interesting alternative to more traditional
indicator organisms such as E. coli and Enterococci.5 Since they are strict anaerobes, they are indicative of
recent fecal contamination when found in water systems. This is a particularly strong reference point when
trying to determine recent outbreaks in fecal pollution. They are also more abundant in feces of warm-
blooded animals than E. coli and Enterococci.
The Human Bacteroidetes IDTM service is designed around the principle that fecal Bacteroidetes are found
in large quantities in feces of warm-blooded animals.3,5,6,7,8 Furthermore, certain strains of Bacteroidetes
have been found to be associated with humans.3,6 As such, these bacterial strains can be used as
indicators of human fecal contamination.
Accuracy of the results is possible because the method amplifies DNA into a large number of small copies
of the gene biomarker of interest. This is accomplished with small pieces of DNA called primers that are
complementary and specific to the unique B. dorei DNA sequence. Through a heating process called
thermal cycling, the double stranded DNA is denatured, hybridized to the complementary primers and
amplified to create many copies of the DNA fragment desired. If the primers are successful in finding a site
on the DNA fragment that is specific to the B. dorei DNA sequence, then billions of copies of the DNA
fragment will be available and detected in real-time. The accumulation of DNA product is plotted as an
amplification curve by the qPCR software. The absence of an amplification curve indicates that the B. dorei
gene biomarker is not detected in the water sample because it is either not present or present at
concentrations below the analytical detection limit.
To strengthen the validity of the results, additional tests targeting other high-ranking, human-associated
Bacteroidetes species should be performed, such as
Human Bacteroidetes IDTM Species: B. stercoris,
Human Bacteroidetes IDTM Species: B. fragilis, and
Human Bacteroidetes IDTM Species: B. thetaiotaomicron.
1Boehm, A., Fuhrman, J., Mrse, R., Grant, S. Tiered approach for identification of a human fecal pollution source at a recreational beach:
case study at Avalon Bay, Catalina Island, California. Environ Sci Technol. 2003 37: 673680.
2Bakir, M., Sakamoto, M., Kitahara, M., Matsumoto, M., Benno, Y. Bacteroides dorei sp. nov., isolated from human faeces. Int. J. Syst. Evol.
Microbiol. 2006 56: 16391641.

3 Bernhard, A., Field, K. A PCR assay to discriminate human and ruminant feces on the basis of host differences in Bacteroides-Prevotella
genes encoding 16S rRNA. Appl. Environ. Microbiol. 2000b 66: 4571-4574.
4Ahmed, w., Masters, N., Toze, S. Consistency in the host specificity and host sensitivity of the Bacteroides HF183 marker for sewage
pollution tracking. Lett. Appl. Microbiol. 2012 55: 283-289.

5 Scott, T., Rose, J., Jenkins, T., Farrah, S., Lukasik, J. Microbial Source Tracking: Current Methodology and Future Directions. Appl. Environ.
Microbiol. 2002 68: 5796-5803.

6 Bernhard, A., Field, K. Identification of nonpoint sources of fecal pollution in coastal waters by using host-specific 16S ribosomal DNA
genetic markers from fecal anaerobes. Appl. Environ. Microbiol. 2000a 66: 1587-1594.
7 Fogarty, L., Voytek, M. A Comparison of Bacteroides-Prevotella 16S rRNA Genetic Markers for Fecal Samples from Different Animal
Species. Appl. Environ. Microbiol. 2005 71: 5999-6007.

8 Dick, L., Bernhard, A., Brodeur, T., Santo Domingo, J., et al. Host Distributions of Uncultivated Fecal Bacteroidales Bacteria Reveal Genetic
Human Bacteroidetes IDTM: EPA Developed Assay
The Human Bacteroidetes IDTM: EPA Developed Assay service targets a functional gene biomarker in
Bacteroidales-like anaerobic bacteria that is present in high concentrations in the human gut. The U.S
Environmental Protection Agency (U.S. EPA) was the first to target the biomarker using quantitative
Polymerase Chain Reaction (qPCR) technology in order to detect ground and surface waters impacted by
human fecal pollution.1 Since it's development, the assay has been used succesfully around the U.S to
identify fecal pollution originating from human sources, such as sewage and septage wastewaters.
The U.S. EPA Developed assay has been shown to be highly associated with human fecal pollution. It has
successfully been validated in multiple nationwide studies using at least 300 individual reference fecal
material from 22 different animal species known to commonly contaminate environmental waters.1,2 A
reported 99.2% specificity to human fecal material makes this one of the leading assays to confirm the
presence of fecal contamination that is of human origin.1 The Bacteroidales-like bacteria is widely
distributed. It was detected in 100% of hundreds of sewage and human reference fecal samples collected
from more than 20 human populations, making it highly sensitive. Internal validations have also been
conducted on hundreds of wastewater, human and animal host fecal samples archived in the Source
Molecular fecal bank.
Fecal anaerobic bacteria are considered for several reasons an interesting alternative to more traditional
fecal indicator organisms such as E. coli and Enterococci.3 Since they are strict anaerobes, they are
indicative of recent fecal contamination when found in water systems.3 This is a particularly strong
reference point when trying to determine recent outbreaks in fecal pollution. They are also more abundant
in feces of warm-blooded animals than E. coli and Enterococci.
The Human Bacteroidetes IDTM: EPA Developed Assay service is designed around the principle that
fecal Bacteroidales-like bacteria are found in large quantities in feces of warm-blooded animals.4,5
Furthermore, certain strains have been shown to be associated with humans.4,5 As such, these bacterial
strains can be used as indicators of human fecal contamination. An advantage of the Human Bacteroidetes
IDTM service is that the entire portion of water sampled is filtered to concentrate bacteria. As such, this
method avoids the randomness effect of culturing and selecting bacterial isolates. This is an advantage for
highly contaminated water systems with potential multiple sources of fecal contamination.
Accuracy of the results is possible because the method amplifies DNA into a large number of copies of the
gene biomarker of interest. This is accomplished with small pieces of DNA called primers that are
complementary and specific to the gene biomarker. Through a heating process called thermal cycling, the
double stranded DNA is denatured, hybridized to the complementary primers and amplified to create many
copies of the DNA fragment. If the primers are successful in finding a site on the DNA fragment that is
specific to the human-associated biomarker, billions of copies of the DNA fragment will be available and
detected in real-time. The accumulation of DNA product is plotted as an amplification curve by qPCR
software. The absence of an amplification curve indicates that the gene biomarker is not detectable in the
water sample either because it is not present or present at concentrations below the analytical detection
limit.
To strengthen the validity of the results, additional tests targeting other high-ranking, human-associated
Bacteroidetes species should be performed, such as
Human Bacteroidetes IDTM Species: B. dorei,
Human Bacteroidetes IDTM Species: B. fragilis, and
Human Bacteroidetes IDTM Species: B. stercoris
1 Shanks, O., Kelty, C., Sivaganesan, M., Varma, M. and Haugland, R. Quantitative PCR for Genetic Markers of Human Fecal Pollution. Appl.
Environ. Microbiol. 2009 75: 5507-5513.
2 Layton, B., Cao, Y., Ebentier, D., Hanley, K., Ballest, E., Brando, J., et al. Performance of Human Fecal Anaerobe-Associated PCR-Based
Assays in a Multi-Laboratory Method Evaluation Study. Water Research. 2013 In Press.

3 Scott, T., Rose, J., Jenkins, T., Farrah, S. and Lukasik, J. Microbial Source Tracking: Current Methodology and Future Directions. Appl.
Environ. Microbiol. 2002 68: 5796-5803.
4 Bernhard, A., Field, K. Identification of nonpoint sources of fecal pollution in coastal waters by using host-specific 16S ribosomal DNA
genetic markers from fecal anaerobes. Appl. Environ. Microbiol. 2000a 66: 1587-1594.
5 Bernhard, A., Field, K. A PCR assay to discriminate human and ruminant feces on the basis of host differences in Bacteroides-Prevotella
genes encoding 16S rRNA. Appl. Environ. Microbiol. 2000b 66: 4571-4574.
APPENDIX E
Phone: (813)630-9616
Fax: (813)630-4327
March 20, 2017
Kevin Riskowitz
City of St. Petersburg
1635 3rd Ave. N.
Saint Petersburg, FL 33713
RE: Workorder: T1703922 Arcadis City of St. Pete
Dear Kevin Riskowitz:
Enclosed are the analytical results for sample(s) received by the laboratory on Monday, March 06, 2017. Results reported herein conform
to the most current NELAC standards, where applicable, unless otherwise narrated in the body of the report. The analytical results for the
samples contained in this report were submitted for analysis as outlined by the Chain of Custody and results pertain only to these samples.
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Enclosures
Report ID: 474531 - 323790 Page 1 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
SAMPLE SUMMARY
Workorder: T1703922 Arcadis City of St. Pete
Lab ID Sample ID Matrix Date Collected Date Received
T1703922001 T170306 LK-1 Water 3/6/2017 09:00 3/6/2017 14:15

T1703922002 T170306 LK-2 Water 3/6/2017 09:45 3/6/2017 14:15
T1703922003 T170306 LK-3 Water 3/6/2017 10:45 3/6/2017 14:15
T1703922004 T170306 LK-4 Water 3/6/2017 11:20 3/6/2017 14:15
Report ID: 474531 - 323790 Page 2 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

Sample ID: T170306 LK-1 Date Collected: 03/06/17 09:00
RegLmt
Adjusted Adjusted
Microbiology
Coliform Total 200 #/100 mL 10 10 10 3/6/2017 17:30 T

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.16 mg/L 1 0.10 0.02 3/9/2017 11:22 T



SM10200H,Water
Pheophytin A 27 mg/m3 1 1.0 1.0 3/15/2017 11:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:44 T
Nitrite 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:44 T
Report ID: 474531 - 323790 Page 3 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.12 mg/L 1 0.10 0.02 3/9/2017 11:22 T



SM10200H,Water

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:45 T
Nitrite 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:45 T
Report ID: 474531 - 323790 Page 4 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.13 mg/L 1 0.10 0.02 3/9/2017 11:22 T



SM10200H,Water

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:46 T
Nitrite 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:46 T
Report ID: 474531 - 323790 Page 5 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS

RegLmt
Adjusted Adjusted
Microbiology

MF,SM9222D,Water
WET CHEMISTRY
Ammonia (N) 0.12 mg/L 1 0.10 0.02 3/9/2017 11:22 T



SM10200H,Water
Chlorophyll C 16 mg/m3 1 1.0 1.0 3/15/2017 11:30 G

Solids,SM2540C


SM4500NO3F,Water
Nitrate 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:47 T
Nitrite 0.18 U mg/L 1 0.20 0.18 3/8/2017 09:47 T
Report ID: 474531 - 323790 Page 6 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
ANALYTICAL RESULTS QUALIFIERS
PARAMETER QUALIFIERS
U The compound was analyzed for but not detected.
I The reported value is between the laboratory method detection limit and the laboratory practical quantitation limit.
B Results based upon colony counts outside the acceptable range.
LAB QUALIFIERS
G DOH Certification #E82001(AEL-G)(FL NELAC Certification)
T DOH Certification #E84589(AEL-T)(FL NELAC Certification)
Report ID: 474531 - 323790 Page 7 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
QC Batch: WCAt/7450 Analysis Method: SM 4500NO3-F

QC Batch Method: SM 4500NO3-F Prepared:
Blank Reporting
WET CHEMISTRY
Nitrate mg/L 0.18 0.18 U
Nitrite mg/L 0.18 0.18 U
Spike LCS LCS % Rec

WET CHEMISTRY
Nitrate mg/L 1 1.0 101 90-110
Nitrite mg/L 1 1.0 100 90-110

WET CHEMISTRY
Nitrate mg/L 0.26 1 1.3 1.2 100 98 90-110 1 10
Nitrite mg/L 0 1 1.1 1.1 110 110 90-110 0 10

QC Batch Method: EPA 350.1 Prepared:
Blank Reporting
WET CHEMISTRY
Ammonia (N) mg/L 0.02 0.02 U
Report ID: 474531 - 323790 Page 8 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Spike LCS LCS % Rec

WET CHEMISTRY
Ammonia (N) mg/L 0.5 0.47 93 90-110

WET CHEMISTRY
Ammonia (N) mg/L 1.3 1 2.4 2.4 105 105 90-110 0 10

WET CHEMISTRY
Ammonia (N) mg/L 0.39 1 1.5 1.5 110 107 90-110 2 10
QC Batch: WCAt/7469 Analysis Method: SM 2540 C

QC Batch Method: SM 2540 C Prepared:
Blank Reporting
WET CHEMISTRY
Total Dissolved Solids mg/L 10 10 U
Spike LCS LCS % Rec

WET CHEMISTRY
Total Dissolved Solids mg/L 660 680 102 75-125
Report ID: 474531 - 323790 Page 9 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Original DUP Max

WET CHEMISTRY
Original DUP Max

WET CHEMISTRY
Associated Lab Samples: T1703922001
Blank Reporting
WET CHEMISTRY
Spike LCS LCS % Rec

WET CHEMISTRY
Original DUP Max

WET CHEMISTRY
Total Suspended Solids mg/L 1.0 1.0 0 10
Report ID: 474531 - 323790 Page 10 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Associated Lab Samples: T1703922002, T1703922003, T1703922004
Blank Reporting
WET CHEMISTRY
Spike LCS LCS % Rec

WET CHEMISTRY
Original DUP Max

WET CHEMISTRY
Total Suspended Solids mg/L 14 13 6 10
Original DUP Max

WET CHEMISTRY
Total Suspended Solids mg/L 1.0U 1.0 0 10
QC Batch: WCAg/4404 Analysis Method: EPA 365.3
QC Batch Method: EPA 365.3 Prepared: 03/09/2017 15:22
Blank Reporting
WET CHEMISTRY
Total Phosphorus (as P) mg/L 0.005 0.005 U
Report ID: 474531 - 323790 Page 11 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Spike LCS LCS % Rec

WET CHEMISTRY
Total Phosphorus (as P) mg/L 0.1 0.100 100 80-120
MATRIX SPIKE & MATRIX SPIKE DUPLICATE: 2294607 2294608 Original: J1702387024

WET CHEMISTRY

WET CHEMISTRY

Blank Reporting
WET CHEMISTRY
Spike LCS LCS % Rec

WET CHEMISTRY
Report ID: 474531 - 323790 Page 12 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327

WET CHEMISTRY

WET CHEMISTRY
QC Batch: WCAg/4435 Analysis Method: SM 10200 H

QC Batch Method: SM 10200 H Prepared:
Blank Reporting
WET CHEMISTRY
Chlorophyll A mg/m3 1.0 1.0 U
Chlorophyll B mg/m3 1.0 1.0 U
Chlorophyll C mg/m3 1.0 1.0 U
Pheophytin A mg/m3 1.0 1.0 U
Original DUP Max

WET CHEMISTRY
Chlorophyll A mg/m3 81 81 1 20
Chlorophyll B mg/m3 90 93 3
Chlorophyll C mg/m3 9.3 9.9 6
Pheophytin A mg/m3 27 25 8
QC Batch: MICt/2598 Analysis Method: SM 9222D
Report ID: 474531 - 323790 Page 13 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Blank Reporting
Microbiology
Coliform Fecal #/100 mL 1 1 U
QC Batch: MICt/2609 Analysis Method: SM 9222 B (MF)

QC Batch Method: SM 9222 B (MF) Prepared:
Blank Reporting
Microbiology
Coliform Total #/100 mL 1 1 U
Report ID: 474531 - 323790 Page 14 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Analysis
T1703922001 T170306 LK-1 SM 4500NO3-F WCAt/7450

T1703922002 T170306 LK-2 SM 4500NO3-F WCAt/7450
T1703922003 T170306 LK-3 SM 4500NO3-F WCAt/7450
T1703922004 T170306 LK-4 SM 4500NO3-F WCAt/7450
T1703922001 T170306 LK-1 EPA 350.1 WCAt/7455

T1703922002 T170306 LK-2 EPA 350.1 WCAt/7455
T1703922003 T170306 LK-3 EPA 350.1 WCAt/7455
T1703922004 T170306 LK-4 EPA 350.1 WCAt/7455
T1703922001 T170306 LK-1 SM 2540 C WCAt/7469

T1703922002 T170306 LK-2 SM 2540 C WCAt/7469
T1703922003 T170306 LK-3 SM 2540 C WCAt/7469
T1703922004 T170306 LK-4 SM 2540 C WCAt/7469
T1703922001 T170306 LK-1 SM 2540D WCAt/7473
T1703922002 T170306 LK-2 SM 2540D WCAt/7474

T1703922003 T170306 LK-3 SM 2540D WCAt/7474
T1703922004 T170306 LK-4 SM 2540D WCAt/7474
T1703922001 T170306 LK-1 EPA 365.3 WCAg/4404 EPA 365.3 WCAg/4421

T1703922001 T170306 LK-1 Copper Sulfate Digestion WCAt/7499 EPA 351.2 WCAt/7509
T1703922001 T170306 LK-1 SM 10200 H WCAg/4435

T1703922002 T170306 LK-2 SM 10200 H WCAg/4435
Report ID: 474531 - 323790 Page 15 of 17
3004.1.0.0
Phone: (813)630-9616
Fax: (813)630-4327
Analysis
T1703922003 T170306 LK-3 SM 10200 H WCAg/4435

T1703922004 T170306 LK-4 SM 10200 H WCAg/4435
T1703922001 T170306 LK-1 SM 9222D MICt/2598

T1703922002 T170306 LK-2 SM 9222D MICt/2598
T1703922003 T170306 LK-3 SM 9222D MICt/2598
T1703922004 T170306 LK-4 SM 9222D MICt/2598
T1703922001 T170306 LK-1 SM 9222 B (MF) MICt/2609

T1703922002 T170306 LK-2 SM 9222 B (MF) MICt/2609
T1703922003 T170306 LK-3 SM 9222 B (MF) MICt/2609
T1703922004 T170306 LK-4 SM 9222 B (MF) MICt/2609
Report ID: 474531 - 323790 Page 16 of 17
3004.1.0.0
Monday, March 20, 2017 11:31:12 AM
Page 17 of 17
Arcadis U.S., Inc.
3109 West Dr. Martin Luther King Jr. Boulevard

Suite 350
Tampa, Florida 33607
Tel 813 903 3100
Fax 813 903 9115
www.arcadis.com

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VERSION CONTROL, OPTIONAL

Sample Collection ................................................................................................................................4-2

5 Conclusions and Recommendations ...................................................................................................5-4

Graphic 1: AccuWeather Temperatures for Saint Petersburg .........................................5-4

Appendix C AEL January Sample Report

4 SAMPLE COLLECTION AND ANALYSIS

5 CONCLUSIONS AND RECOMMENDATIONS

Graphic 1: AccuWeather Temperatures for Saint Petersburg

Coffee Pot Bayou

Miles 1 inch = 20,833 feet

Water Sampling for Pelican January 2017

FWC Mortality Reports

Miles 1 inch = 4 miles

Pelican Mortality Study FWC Wildlife Mortality Reports March 2017

0 250 500 1,000 1,500 2,000

Feet 1 inch = 1,000 feet

Water Sampling for Pelican March 2017

Sample and Water Quality Location

0 120 240 480

Source: Esri, DigitalGlobe, GeoEye, Earthstar Geographics, CNES/Airbus

Riviera Lake #1 Figure X

1 Stormwater runoff from

Sample Results: To Tampa Bay

1 The cold air and rain caused

2 The movement of the water

2 The lack of oxygen

The bacteria 1 Clostridium Botulinum

RE: Workorder: T1701171 Arcadis St.Pete SW

Dear Kevin Riskowitz:

Report ID: 467256 - 120933 Page 1 of 25

Workorder: T1701171 Arcadis St.Pete SW

Lab ID Sample ID Matrix Date Collected Date Received

T1701171001 T170119-01 Water 1/19/2017 09:15 1/19/2017 17:15

Report ID: 467256 - 120933 Page 2 of 25

Workorder: T1701171 Arcadis St.Pete SW

Lab ID: T1701171001 Date Received: 01/19/17 17:15 Matrix: Water

Analysis Desc: Fecal Coliform Analytical Method: SM 9222D

Analysis Desc: TKN,E351.2,Water Preparation Method: Copper Sulfate Digestion

Analysis Desc: Total Preparation Method: EPA 365.3

Analysis Desc: Chlorophylls, Analytical Method: SM 10200 H

Analysis Desc: Tot Dissolved Analytical Method: SM 2540 C

Analysis Desc: TSS,SM2540D,Water Analytical Method: SM 2540D

Analysis Desc: Nitrate,Nitrite Analytical Method: SM 4500NO3-F

Report ID: 467256 - 120933 Page 3 of 25

Workorder: T1701171 Arcadis St.Pete SW

Lab ID: T1701171002 Date Received: 01/19/17 17:15 Matrix: Water

Analysis Desc: Fecal Coliform Analytical Method: SM 9222D

Analysis Desc: TKN,E351.2,Water Preparation Method: Copper Sulfate Digestion

Analysis Desc: Total Preparation Method: EPA 365.3

Analysis Desc: Chlorophylls, Analytical Method: SM 10200 H

Analysis Desc: Tot Dissolved Analytical Method: SM 2540 C

Analysis Desc: TSS,SM2540D,Water Analytical Method: SM 2540D

Analysis Desc: Nitrate,Nitrite Analytical Method: SM 4500NO3-F

Report ID: 467256 - 120933 Page 4 of 25

Workorder: T1701171 Arcadis St.Pete SW

Lab ID: T1701171003 Date Received: 01/19/17 17:15 Matrix: Water

Analysis Desc: Fecal Coliform Analytical Method: SM 9222D

Analysis Desc: TKN,E351.2,Water Preparation Method: Copper Sulfate Digestion

Analysis Desc: Total Preparation Method: EPA 365.3

Analysis Desc: Chlorophylls, Analytical Method: SM 10200 H

Analysis Desc: Tot Dissolved Analytical Method: SM 2540 C