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My Research Proposal: Ecological Paleolimnology
« on: September 25, 2007, 11:58:06 AM »
Greetings esteemed ladies and gentlemen. I am currently researching and finishing up my research proposal for my biology/history senior composition (a required bachelor's thesis), which will be a year long project starting this semester till late March of next semester. I just finished my research proposal, and I am looking and asking fellow scientists in here to read my proposal and give me any hints and recommendations on things either it be in spelling, in methodology, in terminology, in citation etc. Please please guys, i need as much recommendation as possible. This is merely the proposal, which is the basis of my research--i dont actually begin laboratory and coring analysis until around october-november. By which, i will be spending most of my time in the lab or in the library or in the historical references on the development of the lake. I greatly appreciate any recommendations, i reiterate gain, to anything. I dont care if it is positive or negative, give it to me. I need input. An early thank you to everyone who helps out!!!
 
Here is my proposal essay:
+++++++++++++++++++++++++++++


Historiographic Analysis of Sandy Lake and Eutrophication during the Turn of the
20TH Century: N:P ratios and Metals Identification




 
A. Lorenzo Lucino Jr.
September 21, 2007

Introduction:
   The status of lakes throughout the world has been substantially affected by the onslaught of invasive human activity that has resulted in eutrophication of freshwater habitats, particularly lakes and ponds (Cole, 1983; Burgis and Troughton, 1987; Brock, 47). Numerous studies have procured that the specific limiting nutrients in eutrophic lakes throughout the world are the high levels of Phosphorus and Nitrogen that are given off from the sewage effluents of erected human housings, agricultural wastes from fertilizers, industrial wastes and increased water runoff by the effects of deforestation, atmospheric pollution and groundwater infiltration (Cole, 1977; Apelin et al, 1996; Markham, 1994; Schindler 1977; McMartin, 1994).
Nutrient loading of Phosphorus and Nitrogen in specific lakes and ponds have been quite disastrous for owners in that such occurrence results in the increase of macrophytes and other planktonic organisms such as dinoflagellates or commonly know as Cyanophyta, which are dominant in Phosphorus limited areas; some species of Nostacales, which has nitrogen fixing capabilities, become dominant if reduced Nitrogen becomes a limiting factor (Sommer, 1989).
One other effect of increased nutrient loading in lakes that are flanked by increased human activity is the degradation in water quality by an increase in algal biomass, discoloration and floating debris, which become an aesthetic problem for summer resorts that depend much on lake clarity( Becker and Neitzel, 1989; Cole, 1983). Additionally, eutrophic freshwaters with persistent algal biomass and debris become a health hazard for people whose living depends on a particular water body (Flemer, 1972; Hsiang-Te and Long-Gen, 1991); this will eventually lead to restrictions on water use such as drinking water, fish kills and odor. Another indicator of eutrophication in a freshwater habitat would be the Secchi disk visibility of particular lakes (Brock, 1985, Schindler, 1980; Smith, 1982); in clean and pristine lakes that are termed oligotrophic, Secchi disk visibility is high due to the considerable low algal biomass; however in eutrophic lakes where there is a considerable increase in algal biomass, the Secchi disk visibility is significantly lower than that of oligotrophic lakes. Thus an excessive eutrophication problem in a particular resort with limited recreational space becomes a financial problem (Schindler, 1978; Schindler 1977).
   These specific indicators are effective in deducing whether or not a particular freshwater habitat is eutrophic or oligotrophic. Considering the fact that Phosphorus is critical for the growth of algal biomass and that varying Nitrogen levels are responsible for the presence of nitrogen fixing taxa; these two limiting factors are pivotal for understanding the eutrophication processes of lakes and other freshwater habitats. The comprehension of the paleolimnology of freshwater habitats is essential in that one can analyze the Phosphorus to Nitrogen levels during the primordial epoch before the advent of human invasive presence and compare those with levels during the era of human colonization and industrialization (Aplin et al, 1996). The collection of these data provides a better understanding of how industrialization and human activity in different regions of the globe affects the surrounding freshwater habitat via eutrophication. Other effects of industrialization that will be studied is the metallic sediments given off by coal emissions by coal plants. One in particular that will be studied was the coal plant that released its emissions in Sandy Lake, Pennsylvania. Proper analysis of paleolimnology will link eutrophication of Sandy Lake during the late 19th century and early 20th century to coal metal emissions.
Background Information:
   The problems that come with the advent of heavy human colonization near lake areas and the rise of industrialization has been a contributing factor to the quality of lakes in the world (Markham, 1994). Ranging from the classical 18th century Great Industrial Revolution in Europe and later reverberating the same scheme in newly formed independent states in North America, Asia, Latin America and Africa (Hood and Porro, 2002; Magoc, 2006).
   To fully understand the magnamity of the situation, it’s necessary to analyze the situation prior to the arrival of man in certain fresh water habitats, particularly in North America (Burgis and Morris, 1987). In one analysis of the paleolimnologic implications Linsley Pond and surrounding water beds in New England was that after the retreat of Pleistocene ice, these lakes were primarily oligotrophic for a short time before its phase towards eutrophy. This particular stage in the two lakes persisted in dynamic equilibrium for a millennia or so. However when one compares Linsley Lake with Potato Lake in Arizona, the main difference would be Potato Lake’s response to ameliorating climate in depositing organic oozes typical of eutrophy. In some data (Cole, 1975; Cole 1983), there was an increase in 60% ignition loss, yet a sudden decline in organic percentage; which correlates with the increased erosion of the drainage basin’s inorganic content in response to human agriculture and logging activities (Cole, 1975).
   One thing that both Potato Lake and Linsley Pond both share, despite their geographic distance was that there was for a time, a long period of equilibrium in lake water. The nutrients entering the then undisturbed lakes received from yearly runoffs; which correlates with constant annual production and the bottom deposit’s constant accumulation, showcasing an example of trophic equilibrium (Cole, 1975), which reflects the status of the entire edaphic-climatic-morphologic system (Welch, 1992).
   Changes in the trophic levels of these two specific lakes occurred right after the arrival and the establishment of European settlers in North America. Such activities that can be accountable for these changes would be forest cutting, agriculture; such disturbance resulted in increased nutrient input in the lakes. Studies also showed that even in Arizona, the loss of natural shrubby cover, called chaparral, to grass resulted in edaphic nutrient erosion, in particular nitrate, to the surrounding freshwater watershed. This of course is an effect of mass human agriculture (Howarth et al, 1988).
Such examples of increased nutrient loading and the historical reasoning for such Phosphorus and Nitrogen increase is universal as the same occurrence happened in Japan starting in the Meiji Restoration epoch, which occurred in the late 19th century after the deposition of the last Tokugawa Shogunate and the rise of the Imperial Privy Council as well as the immediate industrial revolution that took hold of Japan during its industrialization (Apelin et al, 1996). Such consequences of excessive human development and industrialization has been quite observed in other states in Asia that industrialized quite immediately after independency such as the People’s Republic of China, South Korea, the Philippines, The Federation of Malaysia, Indonesia, Vietnam, and India . Other regions of the world that experienced the same effects as those observed in North American as well as European industrialization were the states of South Africa, New Zealand, Australia, Liberia, Israel, Brazil, Mexico and Chile to name a few (Hood and Porro, 2002).
Specific examples of how industrialization and excessive human activities have resulted in increased nutrient loading such as Nitrogen and Phosphorus levels in fresh water bodies and thus titillated eutrophy is seen in Australia’s Lake Burragorang. What happened in the lake was that there was a great fire near the Blue Mountains in 1968 and showed that there was increased Phosphorus being washed into the reservoir, which was responsible for algal blooms, which then tainted and affected water supplies for the surrounding community around the lake (Aplin et al, 1996). Metallic substance can also be determined in its contribution to eutrophication by use of core analysis.
Additionally, there has been other problem in Australia pertaining to agricultural and pastoral use, which has observed in the state of New South Wales in which large amounts of fertilizers that were used on crops reached streams and lakes, where it aids in eutrophication. In one specific example, a toxic blue green algae bloom of cyanobacteria, caused by high amounts of phosphorus, polluted over 1000km of the Darling River in New South Wales (Aplin et al, 1996). Another example of eutrophication via industrial, commercial and agricultural use would be in Shanghai’s Dianshan Lake, which is a major drinking water source as well as subsistence supplier of fish for the people in that region. Due to the heavy deforestation around Dianshan Lake during the 1970s as well as the heavy industrialization occurring now, there has been an observable increase in Phosphorus levels in the lake, which greatly contributed to the eutrophication of Dianshan Lake (Hsiang-Te and Long-Gen, 1991).
The Nature of Phosphorus and Nitrogen:
   Phosphorus is critical to all life, as it is universally observed in ATP processes (Adenosine triphosphate)as carrier of energy and the fact that nucleotides have phosphate groups and thus illustrates how nucleic acids as well as all living things’ need for phosphorus. Phosphorus is also a scarce element as compared to other principal atoms within living organisms such as carbon, hydrogen, oxygen, nitrogen and sulfur. An example of Phosphorus’ (Cole, 1978; Cole 1983) indicative nature in bringing life within freshwater habitat via the eutrophication process is how the turnover rates reflect the amount of phosphate in comparison to algal demand. However, due to increased human activity, the nature of Phosphorus has resulted in ‘over enrichment’ of lakes so to say via the process of Eutrophication, which occurs when sewage and other pollutants associated with human industrialization and activity bring excess amount of phosphorus back to the lakes, thus adversely increasing algal growth, and deoxygenating the fresh water region directly.
   Nitrogen is yet another important factor that is indicative of eutrophication in that its presence is correlated with high percentages of nitrogen fixing bacteria such as Rhizobium (Cole, 1983), which are able to use nitrogenase, which reduces nitrogen into an ammonium ion, which then is then used by photosynthetic primary producers. Studies have recorded that O. rubescens are pivotal in foretelling whether or not a freshwater habitat is going through eutrophication, because the species of nitrogen limited and does not appear in lakes until Nitrogen content does arise (Cole 1983; Smith, 1982).
Significance and Project Statement:
The advent of industrialization in the third world as well as in the growth of human development in modernized states has led to increased waste deposits and industrial effluents in what was once trophically equilibrium lakes and influencing the culture of eutrophy (Stoermer and Smol, 1999; Magoc, 2006; Cooke et al, 1983). The increase deposits of Phosphorus and Nitrogen has resulted in growth of photsynethic algae, increase macrophytes, as well as the odor, and overall unaesthetic nature of a eutrophic lake. The presence of nitrogen (nitrate/nitrite) and phosphorus in the lakes will directly explain the presence of nitrogen fixing bacteria as well as phosphorus limited algae. The sampling of water from different lakes will allow testing for phosphorus and nitrogen; the collection of these data provides a better understanding of how human activity in this region of Northwestern Pennsylvania has affected the freshwater habitat via eutrophication and thus allows state environmental agencies to understand what limiting factors are in Lakes Pleasant as well as Conneaut and apply appropriate procedure to alleviate it. 
Methods:
   So far the best analysis of NO3-N in the water is to reduce nitrate in alkaline buffered solution to nitrite by having the sample go through a column of copperized cadmium metal fillings. The measurement of nitrite is observed by use of diazotization method that will eventually lead to a stable pink azo dye with an absorbance up to about 500µg NO3-N or NO2-N/liter. Concentrations of nitrite can be obtained prior to reduction of NO3 to NO2 via the diazotization technique. The cadmium reduction method will need glass columns for effective analysis; this method will allow analysis of about eight samples per hour per column.
   To begin the analysis of the Nitrogen levels of a particular eutrophic lake, one would first need to prepare duplicate aliquots of nitrate standards to yield 50 ml from each Cd-Cu reduction column. The concentrations used should approximate those of the samples to be analyzed and should have lower and higher concentrations. Then take 50 ml of distilled water, nitrate standards, and samples into graduated cylinders; then add 5 ml of buffer solution and then thoroughly mix. Afterwards, add 10 ml of the buffered sample to the column and then discard the effluent. Finish adding the remaining buffered solution to the column in the cylinder and collect the 2 ml of effluent in the same cylinder, rinse the walls and then shake the cylinder. Afterwards collecting 25 ml of the column effluent, making sure to carry a water blank and a standard solution for each column used. Immediately, add about .5 ml of the sulfanilamide solution to the 25 ml sample of effluent from the column and then mix for 5-8 minutes, afterwards then add in 0.5ml of the naphthyl ethylenediamine solution and mix immediately. Allow the mixture to sit between 10 minutes to 2 hours; afterwards then measure the extinction coefficient at wavelength of 543 nm of the solution in a 1 cm cell. Use distilled water as a reference. If the extinction coefficient is more than 1.2 then dilute it by one half with distilled water and then remeasure. If the samples of the water have a visible coloration, a sample blank without the addition of naphthyl ethylenediamine reagent will be processed and obtain the extinction values of the absorbance of distilled water plus reagents (the blank), the absorbance of samples without naphthyl ethylenediamine reagent (if brown coloration) and the absorbance of standards or samples plus reagents. Afterwards, then prepare a standard curve of OD vs concentration for the corrected standards (ODcorr = ODs –ODb). The sample concentrations can then be read directly from the graph; if the standard curve is linear, as it should be, the concentration samples can be calculated by a unit extinction factor (F):
F= Standard concentration (ug NO3 – N/liter)
      ODs of standard – ODb
Afterwards, then use: ug NO3 –N/liter = F[ODs  of sample – (ODb + ODo)]
Then determine the reduction efficiency of the Cd-Cu columns by determining the extinction coefficient of a 100 ug NO3-N/ liter standard and a 100 ug NO2-N/liter standard. If the efficiency is not greater than 95%, the columns should be repacked. The nitrogen measured by Cd-Cu reduction technique is a combination of NO3-N and NO2-N in the sample. The NO2-N concentrations of the samples must be determined separately without the reduction procedure, as outlined in the next section, and subtracted from the NO3-N + NO2-N values determined.
Given:    p = NO3-N + NO2-N concentrations
    q = NO2-N concentration
   r = efficiency of reduction column, %
Then ug NO3-N/ liter = p-(100/r)q.
To store the columns, pass about 100 ml of distilled water through the columns, followed by 50 ml of the ammonium chloride buffer solution. Leave the columns stored covered with the solution and cover tightly. To analyze the nitrite nitrogen, duplicate the aliquots of multiple nitrite standards with concentrations of the samples to be analyzed. Then take 50 ml samples of distilled water standards, blank s and samples into graduated cylinders and add about 5.0 ml of ammonium chloride buffer and mix. After 5 minutes of mixing, carefully add 1.0 ml of naphthyl ethylenediamine solution and mix. Allow to sit for 1 hour and after wards, compare the extinction of the solution against distilled water of 543 nm. Then path the lengths of 1 cm, 5cm, and 10 cm for nitrite concentrations of 60-300, 30-60 and 0-30 ug NO2-N/liter, respectively. Use the following calculation:
F= Standard concentration (ug NO3 – N/liter)
      ODs of standard – ODb
Afterwards, then use: ug NO3 –N/liter = F[ODs  of sample – (ODb + ODo)]
   The analysis of Phosphorus comes next, particular concentrations of greater than 10 ug PO4-P/liter will go as follows: obtain water samples of 100 ml and heat to between 15 and 30 degrees Celsius. Properly make sure to measure the absorbance of a sample to obtain turbidity correction. Afterwards, add 10-15 ml of the composite reagent from a 25 ml graduated cylinder and mix thoroughly. Afte 1 hour, measure the extinction coefficient of the solution in a 1 to 10 cm cell at a wavelength of 885 nm. Then recalibrate the photometer to zero using distilled water before measuring the extinction coefficient of the sample. Then measure the absorbance of a reagent blank (ODb). Afterwards, subtract the extinction number of the reagent blank and the turbidity-color correction from the value for sample extinction to obtain a corrected sample extinction. Use the following equation: ODcorr = ODs – (ODb + ODturb).
Then prepare a standard curve by finding the absorbance of for standard solutions, which are diluted from the stock solution. The unit extinction factor (F) for PO4-P can be calculated as:
F = Standard concentration (ug PO4-P/liter)
             ODstd- ODb
And: ug PO4-P/liter = F [ODs-(ODB + ODturb)]
For concentrations less than 10 ug PO4-P/liter, double the sample size and extract the blue complex with an organic solvent. Samples of .25 ml should be stored (as discussed earlier). Pipet 200 ml of water sample  into a 250 ml separatory funnel (remember to keep the temperature between 15 to 30 degrees Celsius) and add about 20 ml of the composite reagent; mix thoroughly. To calculate total phosphorus levels, one has to make sure that there will be reagents, the same ones used for inorganic phosphate analyses. Persulfate solution, which is mixed 5% w/v K2S2O8 in distilled water. Add 16 l of the 5% Persulfate solution to 100 ml samples in 250 ml borosilicate flasks. Then place flasks in a boiling water bath for one hour (or autoclave for one-half hour at 1055 g/cm^2. Allow to cool and then adjust the volume to 120 ml. The liberated PO4-P is analyzed according to the methods used for inorganic P04-P (use procedures for concentrations < 10 ug P04-P/liter). The P04-P standards and reagent blanks will then be placed in identical boiling and volume adjustment procedures. Proceed with computation of a standard curve and calculations of sample concentrations.
To collect metal samples given off by coal during the turn of the 20th century, one would apply the following: Metal samples will be taken by core and metal samples will be digested for analysis with an aqua regia solution (2:1 Seastar nitric acid and hydrochloric acid) and headted to 70 degs C for 8-10 hours. After digestion, Teflon filters will be removed and the sample will be centrifuged to separate undigested material from the metal in the solution. The sample will be split into glass and Teflon vials for analysis of key metals: mercury, arsenic (glass), cadmium, lead and zinc and stored at 4 degs C. All metals samples will be analyzed via ICP-MS ELEMENT (magnetic sector-inductively coupled plasma-mass spectrometer). Cd, Pb, and Zn should be determined with a standard liquid sample introduction system. Isolation of the said metals are indicative of effects of eutrophication and biota during the specific period.

Begin historical research of Sandy Lake---?
 
Timeline
-October/November: Go to Lake Conneaut and Lake Pleasant and take water samples.
-November/December: Take additional samples of water and properly label each
-December: Begin analysis for Nitrogen (nitrite/nitrate)
-December/Jan: Begin analysis for Phosphorus (varying levels; greater or less than ug PO3-P/liter)
-February: Metal samples and isolation; data data data
-Feb: Present data to Professor Ostrofsky
-October-March: Begin historical/archive research or Conneaut and Pleasant Lake in Erie County Historical Society and/or Crawford County Historical Society
-Dec-March: Begin Writing Senior Composition

 
Bibliography

Aplin, Graeme; Mitchelle, Peter; Cleugh, Helen; Pitman, Andrew; Rich, David. 1996. Global Environmental Crises. Oxford: Oxford University Press. 37-38, 59-61, 112 p.

Beadle, L.C. 1974. The Inland Waters of Tropical Africa: An Introduction to Tropical Limnology. London: Longman Group Limited. 175 p.

Becker, C. Dale; Neitzel, Duane A. 1992. Water Quality In North American River Systems. Battelle Press. 190-191 p.

Brock, Thomas. D. 1985. A Eutrophic Lake: Lake Mendota, Wisconsin. New York: Springer-Verlag. 47 p

Burgis, Marry. J and Morris, Pat. 1987. The Natural History of Lakes. Cambridge: Cambridge University Press. 40 p.

Cole, Gerald A. 1975. Textbook of Limnology. The C. V. Mosby Company. 15-18, 246 p.

Cole, Gerald A. 1983. Textbook of Limnology: Third Edition. The C. V. Mosby Company.  320-340 p

Cooke, G. Dennis; Welch, Eugene, B; Peterson, Spencer A; Newroth, Peter. R. 1986. Restoration and Management of Lakes and Reservoirs: Second Edition. Lewis Publishers. 32 p.

Corson, Walter H. 1990. The Global Ecology Handbook: What You Can Do About the Environmental Crisis. Boston: Beacon Press. 267,245-246 p.

Flemer, David. A. 1972. Current Status of Knowledge concerning the Cause and Biological Effects of Eutrophication in Chesapeake Bay. Chesapeake Science 13:144-149.

Holdgate, M. W. 1979. A Perspective of Environmental Pollution. Cambridge: Cambridge University Press. 88 p.

Hood, Charles. H; Porro, Roberto. 2002. Deforestation and Land Use in the Amazon. University Press of Florida. 10-12,162-163, 299-300 p.

Howarth, Robert. W; Marino, Roxanne; Lane, Judith; Cole, Jonathan, J. 1988. Nitrogen Fixation in Freshwater, Estuarine, and Marine Ecosystems. 1. Rates and Importance. Limnology and Oceanography 33(4):669-687.

Hsiang-Te, Kung; Long-Gen, Ying. 1991. A Study of Lake Eutrophication in Shanghai, China. The Geographical Journal 157(1):45-50

Magoc. Chris. J. 2006. Environmental Issues in American History. Connecticut: Greenwood Press. 35-38 p.

Markham, Adam. 1994. A Brief History of Pollution. New York: St. Martin’s Press. 52-61,109 p.

McMartin, Barbara. 1994. The Great Forest of the Adirondacks. New York: North Country Books. 154-157 p.

Mitsch, William J. 1994. Global Wetlands: Old World and New. Amsterdam: Elsevier.

Schindler, D.W. 1977. Evolution of Phosphorus Limitation in Lakes. Science 195(4275): 260-262.

Schindler, D.W. 1978. Factors Regulating Phytoplankton Production and Standing Crop in the World’s Freshwaters. Limnology and Oceanography 23(3):478-486.

Schindler, D.W. 1980. The Effect of Fertilization with Phosphorus and Nitrogen Versus Phosphorus Alone on Eutrophication of Experimental Lakes. Limnology and Oceanography 25(6): 1149-1152.

Smith, Val. H. 1982. The Nitrogen and Phosphorus Dependence of Algal Biomass in Lakes: An Empirical and Theoretical Analysis. Limnology and Oceanography 27(6):1101-1112.

Sommer, Ulrich. 1989. Plankton Ecology. Berlin: Springer-Verlag. 58 p.

Stearns, Peter N. 1993. The Industrial Revolution in World History. Westview Press. 116-126 p.

Stewart, W.D.P; Fitzgerald, G.P; Burris, R.H. 1970. Acetylene Reduction Assay for Determination of Phosphorous Availability in Wisconsin Lakes. Proceedings of the National Academy of Sciences of the United States of America 66(4): 1104-1111.

Stoermer, Eugene F and Smol, John. P. 1999. The Diatoms: Application for the Environmental and Earth Sciences. Cambridge: Cambridge University Press. 128-129 p.

Vitousek, Peter. M; Howarth, Robert. W. 1991. Nitrogen Limitation on Land and in the Sea: How can it occur? Biogeochemistry 13(2):87-115.

Welch, E. B. 1992. Ecological Effects of Wastewater: Applied limnology and pollutant effects. E & FN Spon. 186,64-65 p.

Wetzel, Robert G; Likens, Gene E. 1979. Limnological Analyses. W. B. Saunders Company. 89 p.




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Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #1 on: October 27, 2007, 02:38:13 AM »
HA! This week has been absolutely successful. I'm satisfied. I recently had my oral deliberation on my research topic before Dr. Ostrofsky, PhD., and Dr. Rankin, PhD, both of who are my advisors for my Biology Senior composition. I defended my initiative proposal, and soundly retorted any of their secondary opinions, successfully. I also recieved word from Dr. Ostrofsky that the Pennsylvania Department of Environmental Protection and the Fish and Game Agency has recently reviewed my purpose of research and approved my grant to proceed. A grant of $ 1,800.00 was issued to me to fund my research; coring analysis, usage of the ICP-MS ELEMENT (magnetic sector-inductively coupled plasma-mass spectrometer), which will allow a specific analysis of the said metals, as well as allow the progression of a historiographic comparative of Nitrogen to Phosphorus levels and heavy metals indicatives in the primordial epoch, to the advent of human colonization in the 19th century to the modern epoch.

Last week, I went with the Fish and Game association as well as with my professors to sandy Lake to extract oligotrophic limnologic sediments from a core; and were successful in taking out about 65 centimeters of core--which I and my research subordinates will stratify in conducive 1cm intervals. From the length of the given core, we are able to estimate that we can go back as far as 200-250 years. However, after the metals analysis and algal bloom reciprocation statistics, we will be able to find the exact date via carbon dating via alogrithmic methodology.

---
To begin the analysis of the Nitrogen levels of a particular eutrophic lake, one would first need to prepare duplicate aliquots of nitrate standards to yield 50 ml from each Cd-Cu reduction column. The concentrations used should approximate those of the samples to be analyzed and should have lower and higher concentrations. Then take 50 ml of distilled water, nitrate standards, and samples into graduated cylinders; then add 5 ml of buffer solution and then thoroughly mix. Afterwards, add 10 ml of the buffered sample to the column and then discard the effluent. Finish adding the remaining buffered solution to the column in the cylinder and collect the 2 ml of effluent in the same cylinder, rinse the walls and then shake the cylinder. Afterwards collecting 25 ml of the column effluent, making sure to carry a water blank and a standard solution for each column used. Immediately, add about .5 ml of the sulfanilamide solution to the 25 ml sample of effluent from the column and then mix for 5-8 minutes, afterwards then add in 0.5ml of the naphthyl ethylenediamine solution and mix immediately. Allow the mixture to sit between 10 minutes to 2 hours; afterwards then measure the extinction coefficient at wavelength of 543 nm of the solution in a 1 cm cell. Use distilled water as a reference. If the extinction coefficient is more than 1.2 then dilute it by one half with distilled water and then remeasure. If the samples of the water have a visible coloration, a sample blank without the addition of naphthyl ethylenediamine reagent will be processed and obtain the extinction values of the absorbance of distilled water plus reagents (the blank), the absorbance of samples without naphthyl ethylenediamine reagent (if brown coloration) and the absorbance of standards or samples plus reagents. Afterwards, then prepare a standard curve of OD vs concentration for the corrected standards (ODcorr = ODs –ODb).
The analysis of Phosphorus comes next, particular concentrations of greater than 10 ug PO4-P/liter will go as follows: obtain water samples of 100 ml and heat to between 15 and 30 degrees Celsius. Properly make sure to measure the absorbance of a sample to obtain turbidity correction. Afterwards, add 10-15 ml of the composite reagent from a 25 ml graduated cylinder and mix thoroughly. Afte 1 hour, measure the extinction coefficient of the solution in a 1 to 10 cm cell at a wavelength of 885 nm. Then recalibrate the photometer to zero using distilled water before measuring the extinction coefficient of the sample. Then measure the absorbance of a reagent blank (ODb). Afterwards, subtract the extinction number of the reagent blank and the turbidity-color correction from the value for sample extinction to obtain a corrected sample extinction. Use the following equation: ODcorr = ODs – (ODb + ODturb).Then prepare a standard curve by finding the absorbance of for standard solutions, which are diluted from the stock solution.
To collect metal samples given off by coal during the turn of the 20th century, one would apply the following: Metal samples will be taken by core and metal samples will be digested for analysis with an aqua regia solution (2:1 Seastar nitric acid and hydrochloric acid) and headted to 70 degs C for 8-10 hours. After digestion, Teflon filters will be removed and the sample will be centrifuged to separate undigested material from the metal in the solution. The sample will be split into glass and Teflon vials for analysis of key metals: mercury, arsenic (glass), cadmium, lead and zinc and stored at 4 degs C. All metals samples will be analyzed via ICP-MS ELEMENT (magnetic sector-inductively coupled plasma-mass spectrometer). Cd, Pb, and Zn should be determined with a standard liquid sample introduction system. Isolation of the said metals are indicative of effects of eutrophication and biota during the specific period.

---

Some pictures of today's work.

n52000654_30556740_4213 - My Research Proposal: Ecological Paleolimnology - Science and Research
Ah sweet. The core from Sandy Lake; we exctracted about 69 centimeters of lake sediment; which will allow us to go back at least 100-150 years. Allowing Nitrogen-to-Phosphorus analysis, algal pigment analysis, metals analysis (which im doing my comp on). This is the core concepts of the study of Paleolimnology or Paleooceanography. Well the analysis part, that is.

n52000654_30556741_4526 - My Research Proposal: Ecological Paleolimnology - Science and Research
The core again; different angle.

n52000654_30556743_5187 - My Research Proposal: Ecological Paleolimnology - Science and Research
My work desk :)

n52000654_30556745_5811 - My Research Proposal: Ecological Paleolimnology - Science and Research
The limno core on top of a pedestal, which was created by myself--using paleolimnology tool kits (thanks to the help of Dr. Ostrofsky)

n52000654_30556746_6148 - My Research Proposal: Ecological Paleolimnology - Science and Research
Closer view. Behind the core is the equipment that I brought with me to the limnological site to collect the given core. Overall, I spent about $850.00 of my grant finances just merely to fund and borrow these equipment--so that I would have this particular core. Talk about 'workin for the cream filling' LOL!  ;D

n52000654_30556749_7286 - My Research Proposal: Ecological Paleolimnology - Science and Research
Almost finished with core stratification.

n52000654_30556742_4864 - My Research Proposal: Ecological Paleolimnology - Science and Research
My work bench. This is where I spend 4-7 hours of my day sometimes. Doing archival work; and biochemical reviews for my research.

n52000654_30556748_6924 - My Research Proposal: Ecological Paleolimnology - Science and Research
After working 3 hours of core analysis today, i decided to take a break.....

n52000654_30556747_6566 - My Research Proposal: Ecological Paleolimnology - Science and Research
 :P

That was my day. Now ...its the weekend. Yes! :)



hazel

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #2 on: October 27, 2007, 02:47:22 AM »

Wow! kuyawa nimo dong oi.

I salute you my little bro!  ;D :P

Bambi

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #3 on: October 27, 2007, 02:57:06 AM »
Hello Pare,

I admired your natural talent and being too intelligent...but please, don't forget yourself okey? Do have an extra care and relax more than just as a little bit.  Everything what seems to be important can wait, take your time because  there are still more nicer than only learning for instance.....a candle light dinner with pretty girl of your choice, excellent food, red wine and everything ....the dessert afterwards. Take care!

Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #4 on: October 27, 2007, 03:18:51 AM »
Thanks for the advice, Pare!

Yes, you are right. Too much work and no play is bad for the body, lol. I live by the saying "work hard, party  hard." Pero right now, no time for a girl. Girls will come after studies, hahahaha!

Happy

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #5 on: October 27, 2007, 04:58:34 AM »
Dong taasa ana lagi, its beyond my ability of understanding ;D
"There's no perfect life, but we can let God fill it with perfect moments"

Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #6 on: November 05, 2007, 04:28:31 AM »
Happy, its not that hard at all, most of what I explained in lengthy detail primarily is the procedural steps, the results and data are still in motion right now. Currently, I'm in the drying phase and preparing sediment strata to begin metals analysis, which will begin early this week. More to do..

Happy

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #7 on: November 07, 2007, 03:58:23 AM »
Ok, salamat dong, suwayan nako balik basa kung makaya ba haha

Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #8 on: November 18, 2007, 10:40:25 AM »
Extraction is done. Waiting now for the spectrometer to start working...

Some pictures I took this week--progress is going--

n52000654_30574574_3682 - My Research Proposal: Ecological Paleolimnology - Science and Research
Senior Research Assistant, Holly ( PHD doctoral Student in Conservational Limnology)

n52000654_30574575_4029 - My Research Proposal: Ecological Paleolimnology - Science and Research
Core machine.

n52000654_30582082_8653 - My Research Proposal: Ecological Paleolimnology - Science and Research
Dried stratified limnological sediments (prior to being crushed vis-a-vis mortar and pestal)

n52000654_30582085_9591 - My Research Proposal: Ecological Paleolimnology - Science and Research
Drying Oven

n52000654_30582083_8948 - My Research Proposal: Ecological Paleolimnology - Science and Research

n52000654_30582084_9277 - My Research Proposal: Ecological Paleolimnology - Science and Research

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n52000654_30582088_532 - My Research Proposal: Ecological Paleolimnology - Science and Research

n52000654_30582094_2496 - My Research Proposal: Ecological Paleolimnology - Science and Research
Analysis for dry weight

n52000654_30582097_3469 - My Research Proposal: Ecological Paleolimnology - Science and Research


C2H4

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #9 on: November 18, 2007, 10:52:49 AM »
Dong, I tried to read it para makahatag pud ko'g input,

pero wala nakaya sa akong power, naglabad akong ulo...

it's beyond my comprehension...

with that being said,

say Hi to Dr. Marsh for me

 ;)
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Bambi

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #10 on: November 18, 2007, 11:07:23 AM »
Sus......it is lot of things to do, study and understand, right?  Just looking those apparatus.....nalipong nako. Pastilan.....mo schooling na lang  ko sa pagkamanu-nutho oi!

Pare....you will do it blindfolded and that am 100% sure. Let your course of  studies be involved with fun, okey?

Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #11 on: November 18, 2007, 02:02:34 PM »

You're too humble, 'te. This is nothing for a graduate pharmacist as yourself, but I will give your regards to Dr. Marsh when I see him this monday. He'll ask who tho; what should I say ? ;)

C2H4

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #12 on: November 18, 2007, 02:05:57 PM »

Tell him...your sexy Ate.

LOL

 ;)

Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #13 on: November 18, 2007, 02:30:36 PM »
hahaha! do you want me to your number to him as well? LOL!!!!


;) ;) ;)

C2H4

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #14 on: November 18, 2007, 02:32:49 PM »

NgeH! Hahaha!

Ayaw dong, my banana will make Spam out of me...

 ;D


Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #15 on: November 18, 2007, 02:44:00 PM »
Haha joke ra uy!
But yea its really odd calling Dr. Marsh 'Dr. Marsh' because he's only 25!
Despite his age tho, 'te, he's a pretty well known neurobiologist--he's currently working on neurotoxins--revolving analysis of dopamine levels.

Lorenzo

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Re: My Research Proposal: Ecological Paleolimnology
« Reply #16 on: December 11, 2007, 09:50:31 AM »
Alot of progress has ben ascertained these past weeks; I successfully processed the sediment strata into the CNS (Carbon:Nitrogen: Sulfur) machine---so far the results have correlated the primary test theory. As we increase in depth variable, there is an observable increase in Sulfur pecentage in the soil strata, which can probably be explained due to the metallic effluents given of by the coal industry around the Limnological site in the late 19th century--as well as considering the fact that the research site was the center of a railroad industry during the late 19th century: American Industrial Revolution.
The data results concerning the Carbon and Nitrogen levels have been assimilatory as well, as we go in depth to the core strata, there is a sharp decrease in C:N levels primarily in the 18-45 cm range; above that--and below that--shows a revertion  of C:N normalcy. The core level reaches the 69 cm range, and around that range strata--there is an observable increase in CN levels--indicating pre-settlement dates. The reasons for such a decrease in C:N levels in the 18-45 cm ranges is due to the massive agricultural revolution that took place around the limnological site--and one effect of agriculture has is a reductive effect on C:N levels.

Data results for the CNS analysis have proved positive, however, my data results are being sent to the Dpt. of Limnology for re-evaluation before I actually utilize it as a supportive reference.

Right now tho--I'm currently working on the acid digestion of the sediment strata---which is a prepatory step before I can begin the metals analysis. The first batch of strata have been analyzed, the 2nd batch are currently being analyzed as we speak. I have 3 additional batches to complete before I can begin metals analysis.

I plan to finish the 3rd batch (usually takes 5-6 hours to complete a batch) tomorrow.
The 4th batch will be assessed on Wednesday and the last batch will be assessed on Thursday.

There is so much to do till then.  :-X

Lorenzo

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How to kill time while working in Biology Lab
« Reply #17 on: December 11, 2007, 02:50:42 PM »
So....ever had a feeling of complete boredom while in your work place?
I had that exact feeling today---well aside from working and preparing samples in the Biology lab from 11 AM till 12 Midnight---thats right i was there for 13+ hours--I decided to take some pictures to kill some time. LOL. My only free time or Sanity time. But despite long boring hours---I'm quite satisfied in how progress has been attained.

Some of my little quandaries today...

n52000654_30600683_3014 - My Research Proposal: Ecological Paleolimnology - Science and Research
Sediment strata being heated on hot plate; the mixture is of 0.1 g of sediment (rounded to 10^-4 accuracy), concentrated HCl(hydrochloric acid), HNO3(Nitric Acid), H2O2( Hydrogen Peroxide), DI(Distilled water). The total time it takes to prepare each sample vis-a-vis acid digestion is about 5-6 hours. Per Sample. Its extremely time consuming.

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Another view of the 16 samples (my 2nd batch) that I finished today.

n52000654_30600685_3700 - My Research Proposal: Ecological Paleolimnology - Science and Research
**Not to play at home, lol** Concentrated Nitric Acid---yea--can definitely burn a whole through laytex gloves.  8)

n52000654_30600686_4038 - My Research Proposal: Ecological Paleolimnology - Science and Research
Some of my favorite chemicals, lol.  Concentrated Hydrochloric Acid and Sulfuric Acid. Both are necessary in acid digestion and preparatory steps prior to metals analysis.

n52000654_30600687_4381 - My Research Proposal: Ecological Paleolimnology - Science and Research
35% Hydrogen Peroxide. This is necessary in effervescing the sediment sample; in addition to Hydrogen peroxide, I aliquoted DI (distilled H20) in order to allow hydrocarbonic mixture to liquidate the organic compounds within the Erlenmeyer flasks--its rather interesting as the reaction is rather violent---exothermic radiance.

n52000654_30600691_5750 - My Research Proposal: Ecological Paleolimnology - Science and Research
Ah...yours truly trying to titrate each flask per 20 minute substandard. Its necessary to prevent crystallization from forming, particularly after adding the Nitric Acid.

n52000654_30600692_6095 - My Research Proposal: Ecological Paleolimnology - Science and Research
Live Long and Prosper. LoL! A random moment... :P  8)

n52000654_30600694_6782 - My Research Proposal: Ecological Paleolimnology - Science and Research
Reflux step. This step was immediately after adding H2O2 and HCl. Time it took to finish refluxing was about 2.5 hours. (Though strict regul. requires immediate cancellation after .5 of the intial volume repudiated vis-a-vis evaporative synthesis)

n52000654_30600696_7475 - My Research Proposal: Ecological Paleolimnology - Science and Research
The container tubes---

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Random moment #2

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Random moment #3

Lorenzo

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Reply: My Research Proposal: Ecological Paleolimnology
« Reply #18 on: January 25, 2008, 08:34:25 AM »
5th acid digestion sample in assay.
Conclusively finished 16 sediment strata vis-a-vis acid digestion. 5 more samples to complete before analyzing trace metals.

alleghenypictures5060 - My Research Proposal: Ecological Paleolimnology - Science and Research
Aliquoting 2x 3 mL of HNO3 (concentrated nitric acid)

alleghenypictures5070 - My Research Proposal: Ecological Paleolimnology - Science and Research
A sample, actually this is test tube #256, representing an assay of 45-46 cm depth strata. One of the 300+ samples I've collected--hours have been lost to cases of insufficient effervescence-effect, excessive hydrocarbonic reaction. Very time consuming.

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More to come.


Macky Ferniz

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Reply: My Research Proposal: Ecological Paleolimnology
« Reply #19 on: January 25, 2008, 08:53:19 AM »
I have seen studies of world climate by studying ice and glaciers in Alaska. Time is trapped inside the glacier. They are boring holes and the deeper they got, the older the time. They noticed significant amounts of carbon dioxide released in the atmosphere during the industrial revolution of Europe (coal age). They also detected high amounts of CO2 estimated during the time of Mt. Pinatubo's eruption.

There is also one study that concludes dust from the Sahara destroys corral reefs in the Carrebean.

You have some sophisticated aparatus there Lorenzo.
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