Phytoplankton blooms in the Amazon River plume Studying the trend of phytoplankton blooms in the tropical Atlantic is crucial

in assessing the impact of nutrient input from the Amazon River on primary production and carbon export in the open ocean. The tropical Atlantic is of particular interest because of the vast amount of freshwater input from the Amazon. Population increase and agricultural development along the Amazon River has drastically increased the level of nutrients discharged into the Atlantic Ocean. This source of anthropogenic nutrients has been shown to contribute to the pattern of phytoplankton bloom that occurs seasonally north of South America (Smith et al., 1996). Dynamics and pattern of phytoplankton bloom and fresh water input are observed through monthly chlorophyll-a data from the Moderate Resolution Imaging Spectroradiometer, or Aqua Modis. Newly available satellite surface salinity estimates from the Microwave Radiometer of the Aqarius/SAC-D mission will also allow us to observe the pattern of fresh water input from the Amazon, Chlorophyll a concentration and the area of the three predefined zones are plotted and spatial dynamics observed. The resulting plots show fairly stable trends from year to year, with winter months showing the lowest levels of chlorophyll and a sharp increase in chlorophyll during spring. The increase in phytoplankton mass is assumed to be the result of thermal stratification from the warming of surface water. Seasonal trends for the bloom area and chlorophyll concentration however do differ in their progression. Bloom area hits a minimum level for the winter months and remains at the same level until the spring bloom. Plume area typically hits a minimum level and stay at that level for half of the year, suggesting a core bloom area where stable supply of nutrient is available regardless of season. Previous studies using in situ and satellite ocean color data has demonstrated the relationship between Amazon freshwater input and chlorophyll but used older sensors such as the SeaWiFS.

A large region of sustained seasonal phytoplankton growth exists around the mouth of the Amazon River. Discharge from the Amazon basin introduced an average of 209,000 cubic meters of fresh water per second into the oceans surrounding the mouth of the Amazon from 1973 to 1990 (Molinier et al., 1993). This discharge of freshwater forms a vast plume that extends far from the mouth of the Amazon into the open ocean. This plume of freshwater covers an area of 2 x 106 km2 with a layer of water low in inorganic carbon and salinity (Cooley et al., 2007). Covering approximately 18% of the WTNA the Amazon freshwater plume play a key role in biological cycles in the region (Subramaniam et al., 2007). The seasonal variability of the Amazon River discharge into the Equatorial Atlantic Ocean can be observed using stations at the river. Previous studies has monitored the total discharge by combining data from the Obidos station at the Amazon with stations from two tributaries downstream, the Tapajos and Xingu River. The resulting pattern is characterized by higher values from March to August and less discharges from September to February (Molleri et al., 2010). However it is important to note that the Orinoco River may also contribute to the Amazon River plume observed in the study area. The impact of Orinoco River is out of the scope of this study but its input to the Amazon River plume may be a point of interest in future studies. The freshwater discharge carries vast amounts of nutrients, including nitrate, phosphate, and silicate. Chlorophyll a data has shown that there is sufficient nutrient discharge from the Amazon estuary to sustain high seasonal productivity (Walker-Demaster, 1994; Subramaniam et al., 2007; ). Phytoplankton horizontal and vertical distribution patterns have been shown to be impacted greatly according to freshwater discharge from the Amazon River (Smith Jr. et al., 1996; Muller-Karger., 1988). Previous ship-board studies has concluded that chlorophyll concentrations were greatest in a zone outside of high nutrient, turbulent, and low salinity water

that are closest to the coast, but also shoreward from high salinity, low nutrient far from the coast. Vertical distribution of chlorophyll was greatest in the top 5m of the water column, on top of high salinity, low nutrient waters. (Walker-Demaster, 1994). This distribution reflects the environmental regimes created by the high nutrient freshwater river discharge from the Amazon. Nutrients from the Amazon River increasingly come from anthropogenic sources, primarily from the conversion of vast areas of forests to intensive agriculture. (Filoso et al., 2006) High level of phytoplankton growth is observed to extend well into the deep ocean, not just limited to the coastal areas around the river mouth. It is important to study phytoplankton blooms because phytoplankton are an integral part of the ecosystem, as primary producers and the foundation of the marine food web. Furthermore phytoplankton bloom patterns can be an indication of ecosystem change or perturbation, either as part of a natural cycle or from anthropogenic causes (Paerl et al., 2003). Observing this interaction is also important because phytoplankton blooms are responsible for a large portion of carbon export from the surface of the ocean through the biological pump, having direct impact on removal of CO2 from the atmosphere. (Sarma et al., 2007; Subramaniam et al., 2007) The Amazon plume absorbs approximately 5.8 gC m-2 yr-1 of atmospheric CO2 due to the mixing driven air-sea gradient. This contrasts with the release of 8.2 gC m -2 yr -1 from the whole Atlantic ocean. (Kortzinger, 2003; Takahashi et al., 2002) Furthermore, based on flux estimates of plume-related CO2 sink, the outer Amazon plume alone has been estimated to contribute to 15 6 TgC yr -1 of carbon uptake (Cooley et al., 2007). The Amazon plume thus plays a critical role in the balance of carbon in the region. Due to the fact that a small but growing fraction of nutrient inputs from the Amazon are from anthropogenic sources such as intensive agriculture, this study has the larger implication of the impact on carbon exports from anthropogenic causes that the region may experience in the future.

The significant presence of nitrogen fixing diazotroph plays a key role in nitrogen availability in the region and can greatly impact carbon sequestration. Freshwater from the Amazon has been shown to support diazotroph growth, leading to N2 fixation far from the mouth of the river. (Subramaniam et al., 2007). Near the mouth in low salinity waters, there are enough P, Si, and N to support coastal diatom growth, with little N2 fixation, however as the plume mixes with high salinity-low nutrient water, N is depleted before P and Si, and diazotrophy becomes a significant source of new nitrogen. This source of nitrogen supports diatom species with diazotrophic symbionts, and primary production far off-shore beyond the continental shelf (Subramaniam et al., 2007). The increase in primary production is an important pathway for sequestration of atmospheric CO2 in the Western Tropical North Atlantic. Diazotroph blooms in the WTNA enhance CO2 uptake either directly, or through other autotrophs by providing biologically available nitrogen. Previous ship-based studies have shown that the diazotroph community in the Amazon River plume enhanced the air-sea CO2 disequilibrium by up to 100x. Biological influence from diazotrophs leads to an estimated 15 Tg C yr-1 increase in CO2 uptake in the outer Amazon plume, compared to the influence of mixing alone. Nitrogen fixation by diazotrophs results in net CO2 uptake when traditionally the western tropical north Atlantic was thought to experience CO2 outgassing (Cooley et al., 2007). In studying the characteristics of the phytoplankton plume off the Amazon River, there are several important factors which remain relatively unexplored in this region, principle of which is the interaction between the freshwater plume and the ocean water. Although phytoplankton production is an integral part of marine biological cycles, few time-series of phytoplankton production data sets exist.

The exact spatial pattern and variability of the freshwater plume is unclear up until this point. However salinity data from the Aquarius/SAC-D mission will give us a clear picture for the first time. A time-series study in this area can establish a link between bloom patterns and natural or anthropogenic forcing. Thus it is important to observe the mixing between highnutrient freshwater and the high salinity, low-nutrient ocean water. It is possible that phytoplankton plumes do not follow the extent of freshwater, possibly due to variations in nutrient concentrations in different parts of the plume, or wind driven dynamics affecting the extent of the plume. This comparison will increase the understanding of pattern of phytoplankton plume distribution away from coastal waters and its interaction with low salinity, high nutrient water. Self-shading could also be impacting chlorophyll levels in the study region. As shown from previous iron fertilization experiments, there is sometimes an inverse relationship between chlorophyll and mixed layer depth. High phytoplankton density blocks out available sunlight from lower depths, creating a situation known as self-shading. Self-shading when there is abundant phytoplankton could limit the depth of the euphotic zone and be a limiting factor in phytoplankton growth (De Baar et al., 2005; Krishnamurthy et al., 2006). The impact of selfshading is supported in the region as it has been shown that the Amazon shelf is light limited near shore. Previous studies showing that the coastal areas are dominated by colored dissolved organic matter rather than phytoplankton further supports the possibility of self-shading in the region (Subramaniam et al., 2007; Smith et al., 1996). However, other than self-shading due to high phytoplankton concentration level, phytoplankton production is not limited by the availability of light in the open ocean (Smith et al., 1996). Reduced growth rate resulting from

self-shading in a bloom could also decrease the depletion of nutrients in areas of high phytoplankton concentration, increasing the persistence of a bloom. (Krishnamurthy et al., 2006) These interactions can be observed using newly available data from the Aquarius/SAC-D mission, which monitors global ocean salinity. Chlorophyll a concentration data are obtained from Aqua MODIS (or Moderate Resolution Imaging Spectroradiometer). The reliability of ocean color measurements in the open ocean has been proven by earlier satellite missions such as the CZCS. (Andrew Banks et al. 2011). These studies demonstrated the potential of ocean color measurements for observing chlorophyll concentrations. The early success of the CZCS has lead to subsequent sensors such as SeaWIFS and MODIS. These sensors however, rely solely on the absorption of light by phytoplankton with the assumption that the water is clear and ignores other optically active components of sea water. The empirical relationship between chlorophyll concentration and ocean color from these sensors have generally proven to be strong enough for application in the open ocean, with the exception of specific regions such as the Mediterranean ocean, where waters are more optically complex with a generally higher concentration of colored dissolved organic matter (CDOM). ( Uitz et al.,) However, the assumption that phytoplankton is solely responsible for the absorption of light has led to significant inaccuracies in chlorophyll estimation for more optically complex waters such as coastal waters. (Banks et al., 2011) These waters, classified as Case II, contain other optically active components such as CDOM and nonalgal suspended particulates that breaks down the empirical relationship between ocean color and chlorophyll concentration. (IOCCG, 2000 and Bank et al., 2011; OReilly et al., 1998). Therefore, we focus our study on the open ocean, excluding coastal and shelf waters.

Methods The area of study is located mostly north of the Amazon estuary in the tropical Atlantic Ocean. The study area covers an area approximately 28.0 N to 4.0 S and 70.5 W to 36.5 W and covers 932,688 square kilometers. The use of satellite ocean color data is chosen due to the accessibility of data and for its ability to give high resolution data and provide measurement in areas where in situ measurements are unavailable. The satellite used to obtain chlorophyll-a concentration and salinity measurements are the Aqua MODIS or (Moderate Resolution Imaging Spectroradiometer) and the Aquarius/SAC-D satellite. The resolution used for chlorophyll-a data is 9km obtained from NASAs online ocean color database at http://oceancolor.gsfc.nasa.gov/. The product used is the Aqua MODIS chlorophyll concentration monthly data. All available monthly time series data were used, July 2002 to June 2012. Data was processed using the ENVI imagery analysis software and imported from the ocean color database using the HDF format. We excluded coastal waters bordering the north coast of Brazil. As discussed before, numerous studies have pointed to the overestimation from ocean colors studies on coastal areas where suspended optically active components are present in high quantities. The presence of non-algal suspended particulate matter and colored dissolved organic matters causes significant deviations from true chlorophyll concentrations. The above complications rendered coastal or Case I waters unsuited to be studied via current ocean color techniques. Therefore coastal waters are outside the scope of this study, with the main focus on the impact of nutrients in open ocean Case II waters. Thus coastal waters were excluded in the study area through use of ETOPO 2V2 elevation data obtained from the NOAA (National Oceanic and Atmospheric Administration) data center. Coastal waters were excluded by excluding sea depths less than-100 meters depth from the region of interest. The resulting region of interest or ROI formed the study area.

The study area is further categorized into three different zones based on the chlorophyll concentration. The first zone is classified as open ocean (OOZ), with chlorophyll-a concentration of less than 0.15 mg/m^3. The open ocean zone is not part of the main body of phytoplankton plume, nutrient concentrations in the Open Ocean zone are not high enough to sustain phytoplankton blooms. The plume influenced zone is classified as having chlorophyll-a concentration value of higher than 0.15 mg/m^3 and lower than 1 mg/m^3. In the plume influenced zone (PIZ), nutrient concentrations are high enough to sustain elevated phytoplankton productivity and accounts for the main body of the plume. The intense bloom zone (IBZ) includes waters with the highest concentrations of chlorophyll in the plume. The chlorophyll concentration in the intense bloom zone exceeds 1 mg/m^3. These zones were established based on chlorophyll values that produce clear, contiguous zones in the study area. The PIZ should show the main body of the plume, and the thresholds were made so that the PIZ would be clearly defined with areas not influenced by the Amazon excluded. The average value of each zone were calculated using ENVIs statistics tool by month and the results plotted from September to August of each year starting from 2002, when MODIS data was first available. The area of each zone is also calculated and plotted in the same manner.

RESULTS A colored map of the study area based on chlorophyll concentration shows the extent of a typical phytoplankton bloom during the study period (Figure 1). The plume is represented and each zone clearly shown. The phytoplankton plume in figure 1 is typical of the region in the summer months. High chlorophyll concentrations can be observed in a significant portion of the

study area. From the chlorophyll concentration and area plots we observe definite seasonal trends that are fairly stable and cyclical with a few exceptions. The area of the main body of the phytoplankton plume, or the plume influenced zone maintained a consistent size for the majority of the year, typically from November to April (figure 2). During this non-bloom period, bloom area size is the smallest of the entire year and remains roughly the same size until the spring, with a gradual increase in size starting from April of each year. The increase in bloom area continues until August, where bloom area is typically the highest, this trend was accurate for all years observed except for 2008, which experienced the biggest bloom area in July. (Figure 2) The increase in bloom area is most rapid in July and August, after which bloom area quickly drops off to pre-bloom levels by November of each year. The year 2010 saw the most dramatic increase during bloom season, with bloom area increasing from less than 5,000 square kilometers to over 70,000 square kilometers. The dramatic increase in bloom area in 2010 may indicate nutrient based influence from the Amazon River. (Figure 2) The seasonal change in bloom area was dramatic in the intense bloom zone (figure 3). In the intense bloom zone, seasonal trends in the bloom area were similar to the plume influenced zone, remaining stable from November to April and increasing from May to December. What differentiated the intense bloom zone from the plume influenced zone is the magnitude of increased bloom area for the intense bloom zone. During non-bloom months, the bloom area for the intense bloom zone was typically observed to be below 5,000 square kilometers. However, during bloom months the area with chlorophyll concentration level exceeding 1 mg/m^3 increased by an order of magnitude, a much larger increase than observed for the plume influenced zone, which saw more gradual expansion and shrinkage of the phytoplankton plume. (Figure 3)

Chlorophyll concentration levels for the plume influenced zone exhibit a trend that is cyclical and gradual. Similar to the plume area trends, the chlorophyll concentration for the plume influenced zone are typically at the lowest at November or December or each year. (Figure 4) However, instead of a long period of stable values, the chlorophyll concentration level begins to increase after the lowest point of every year. (Figure 4) The increase is more gradual and hits the highest values on around July of every year. This trend is observed in all of the years. The chlorophyll levels exhibit a cyclical trend that varies from month to month without any long periods of similar values. The intense bloom zones chlorophyll value is much more variable than the plume influenced zone (figure 5). It appears erratic on the plot, but follows the general trend of the plume influenced zone. The year 2011 is notable for a sharp increase in chlorophyll concentration level at a period where chlorophyll concentrations are generally the lowest of the year.

Discussion
The observations confirmed that during the typical spring period from early spring, phytoplankton biomass increased dramatically. Although phytoplankton biomass was not calculated in this study, it can be inferred from the increase in chlorophyll levels in areas typically with low levels of chlorophyll that phytoplankton biomass have multiplied, and thriving in regions previously low in production. The high levels of phytoplankton growth can be attributed to thermal stratification of the water, allowing nutrient input from the Amazon to stay at the surface. Enhanced Stratification of the water can also be caused by the increased flow of freshwater from the Amazon. The less dense water from the Amazon causes freshwater to rest at the top layer of the water column, encouraging phytoplankton production. Based on the fact that the area of phytoplankton growth always maintains a

consistent plume size in non-bloom months and never dips below that minimum area it is likely that the freshwater water mass from the Amazon River is creating density stratification in the water column in an area before the freshwater mixes with the high salinity water. This area of a coherent mass of freshwater likely sits on top of the water column and helped retain nutrients at the surface of the ocean. The abundance of nutrients in this low salinity area is likely to be the reason why the minimum plume area stays consistent even as availability of sunlight decreases in winter months.

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Figure 1. Map of study area showing the Open Ocean (Yellow), plume influenced (Green), and intense bloom (Purple) regions. Note the exclusion of coastal data, other areas in white had no valid chlorophyll data due to cloud cover.

Figure 2. Area in km^2 by months for Plume Influenced and Intense Bloom Region.

Figure 3. Average area of Plume Influenced an Intense Bloom region in km^2 by months across all years.

Figure 4. Average Chlorophyll concentration of Plume Influenced, Intense Bloom, and Open Ocean region in mg/m^3 by months across all years.

Figure 5. Chlorophyll concentration in mg/m^3 by months for Plume Influenced and Intense Bloom region.