by
Renz L. Salumbre
MS Zoology
October, 2008
TABLE OF CONTENTS
Table of Contents......................................................................................................... i
I. Introduction................................................................................................................ 1
B. Global Warming..................................................................................................... 5
III. Methodology............................................................................................................ 11
A. Study Area.............................................................................................................. 11
B. Algal Species......................................................................................................... 11
C. Collection of Data.................................................................................................. 12
D. Data Analysis......................................................................................................... 13
i
D.1 Analysis of Gross Morphology........................................................................... 13
D.4 Computations..................................................................................................... 14
IV. References.............................................................................................................. 15
V. Appendices............................................................................................................... 17
ii
List of Figures
iii
Chapter I
INTRODUCTION
demonstrate that the ozone layer established in the stratosphere retains approximately 95%
to 99% of the sun’s ultraviolet radiation from striking the earth (Sparling, 2002). The sun
produces both visible and invisible rays. Invisible rays known as ultraviolet-B (UVB), which is
290-320 nanometers, cause most of the alleged environmental problems such as the
greenhouse effect and global warming. Affirming this, Kraffert (1998) pointed out that UVB is
Biologically harmful UV-B reaches the earth’s surface in amounts inversely proportional
to the concentration of atmospheric ozone. Some factors that impede UV-B release are urban
air pollution, suspended particulates, aerosols, and ozone in the troposphere, as well as by
stratospheric ozone. UV-B has harmful effects on a wide range of biological systems. It can
cause DNA damage which is proportional to the intensity and duration of exposure;
unfortunately, small, delicate organisms suffer more severe damage than large robust species
(such as humans and other mammals). UV-B, moreover, impairs the growth and
photosynthesis of certain plants and impairs the motility and reproductive capacity of
some of which are already under UV-B stress. Further increases in UV radiation are expected
to cause detrimental effects including disruption of specific food chains in these ecosystems.
phytoplankton will decrease the uptake of CO2 and thus aggravate the greenhouse effect. In
1
short, the ecological effects of increased surface level UV-B radiation, although not fully
predictable, are likely to be widespread and harmful (Hader & Figueroa, 1997).
Macroalgae and sea grasses are important biomass producers in aquatic ecosystems.
In contrast to phytoplankton, most of these organisms are sessile and cannot avoid exposure
to solar radiation at their growth site. Recent investigations showed a pronounced sensitivity
to solar UV-B radiation with the effects being demonstrated throughout the top 10 to 15
meters of the water column. In addition, photosynthesis in the symbiotic algae is impaired,
resulting in reduced organic carbon supply. The succession of algal communities is controlled
by a complex array of external conditions, stress factors and interspecies influence (Hader &
Figueroa, 1997).
The increasing awareness regarding the environmental impact of global warming has
chemistry, physics, or ecology. This said awareness has given rise to the need of more
studies that shall provide a modicum of unmitigated evidence in regard to the destruction of
biological systems. As the ozone layer gets depleted due to various factors, most studies
have focused on the impact of the regressing quality of the ozone layer and how it affects
various species. As such, this paper will take into account the alleged effects of solar UV-B
radiation on algal species in an in vivo setting as opposed to the one originally proposed by
Holzinger & Lütz (1993) whose experiment with ultraviolet irradiation on algae was achieved
in a laboratory setting. Algae present in both Pangasinan and Batangas during the summer
season will be collected while a control group is established and will be maintained.
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Afterwards, the said group of algae will be subjected to a thorough examination of the gross
In this particular study, two representative species of algae both present in Pangasinan
and Batangas will be examined. Simply put, the purpose of this study is to give evidence that
the regressing quality of the ozone layer has a considerable effect on algal communities. The
basis of the study shall consider the visible changes that may have result due to significant
The main objective, therefore, is to formally recognize, if any, the effects of solar UV-B
The specific objectives are the following: first, to identify the degree of destruction
brought about by high levels of solar UV-B radiation on the aforementioned species of algae
in terms of their gross morphology and ultrastructure; second, to compare the gross
morphology and ultrastructure of the above species of algae found in both sites; third, to
corroborate the claim that algae are greatly affected by increasing levels of UV-B radiation;
lastly, to correlate the results on global warming and the destruction of biological systems.
Once this study is accomplished, it is expected that it will add to the growing repository
of knowledge regarding the impact of global warming. It is expected also to help people to be
aware of the increasing destruction brought about by anthropogenic factors which in turn
cause global warming. This study will also corroborate the fact that global warming is altering
3
ecosystems. As the ozone layer is depleted by various factors, plant life or plant structure is
greatly altered in reference to their habitats. This study is also expected to emphasize the
It is also significant to compare the effect of UV-B radiation on the study area (which
includes the Northern and Southern part of Luzon) as well as its effects on the representative
This study will cover, in general, the harmful effects of UV-B radiation on biological
UV-B radiation, as mentioned above, has been observed to be detrimental to plant life
The algal materials that will be scrutinized are Padina minor and Turbinaria conoides
due to the seasonality of algae and the availability of these algae at Bolinao, Pangasinan and
observing the gross morphology and the ultrastructure of these algae with special reference
The abiotic parameters that will be taken into account are the following: pH levels,
Field work shall be accomplished during the summer season while experimentation
4
Chapter II
The stratospheric ozone layer (Figure 1) is practically a shield that protects the earth
from the harmful rays of the sun. The ozone layer, before, is actually thick enough to filter off
the harmful UV-B rays. Solar ultraviolet light splits the ozone’s O3 molecules into O2 and
atomic oxygen, which have a very high potential for recombination, which, in turn, reinforces
the ozone layer; this ozone-making process protects the earth from the cell-damaging effects
of UV-B radiation (UCUSA, 2005). In reality, the biologically harmful ultraviolet radiation
levels reach the earth in very fractional levels. Unfortunately, due to the depleting quality of
the ozone layer and the abundance of atmospheric pollutants, such as fluorocarbons and
nitrogen oxide, the ozone layer cannot give the protection it was made for and has allowed
the transmission of injurious ultraviolet radiation (Acra, Jurdi, Mu’allem, Karahagopian &
Raffoul, 1990).
B. Global Warming
Global warming has been pointed out to be the major consequence of the regression of
quality of the ozone layer. Global warming happens when greenhouse gases are trapped into
the earth, specifically, in the lower atmosphere. It is called as such because when the sun’s
rays enter the earth’s atmosphere, the heat from these rays cannot escape. Global warming
has been considered to be the driving force behind drastic climate changes (ESS, 2000).
5
The sun’s rays reach the earth in the form of light waves. This results to the earth
heating up. The energy that came from the sun is involved in the said heating-up process and
some are re-radiated back into space in the form of infrared waves. It is normal for some of
these infrared waves to be trapped into the earth because it keeps the earth’s temperature
constant enough to be feasible for living. Some problems have arisen in connection with the
increasing number of pollutants that is making the atmosphere thin enough. Usually, these
pollutants come from industrial sources. Moreover, human-caused carbon dioxide and other
greenhouse gases attack the ozone in bulk resulting in the entrapment of large amounts of
infrared waves that should have been re-radiated back into space. The results to the earth’s
Without doubt, global warming has a lasting impact on all life forms. The world is
experiencing a wave of mass extinction of various animals around the world such that the rate
of extinction is a thousand times higher than the normal background rate. The factors that
contribute to this devastating effect are the same factors that contribute to the phenomenon of
climate change; an example of the correlation between these two events is the destruction of
the Amazon rain forest which has resulted in species being driven to their extinction and the
addition of carbon dioxide to the atmosphere. Other effects of global warming are sea level
ris, drastic weather changes and marine life destruction (Gore, 2006).
In a 1998 study, Buccheim provided evidence that ultraviolet radiation is the leading factor
causing coral bleaching and coral mortality. In the study, coral bleaching occurred during
periods of high heat and high penetration of short wavelength ultraviolet radiation. In
connection with this occurrence, Gore (2006) mentioned that during 2005, considered to be
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the hottest year, a massive loss of coral reefs occurred. While earlier, in 1996, the second
hottest year, 16% of the world’s coral reef was lost. Both mention and affirmed the fact that
global warming through increased ocean temperatures mattered greatly in the mortality of
corals.
Not only are corals affected but other marine life forms are affected too. The threat of
marine destruction arises from the uncontrolled growth of carbon dioxide; one-third of whcih
end up sinking into the ocean, thus increasing the acidity of water.
This “chemical change” of the world’s ocean has given rise to dead zones
characteristically devoid of marine life. Others have caused algae to bloom in waters which
are already polluted. These algae can grow so big so as to inhibit not only the natural process
According to Acra et al (1990), the electromagnetic radiation emitted by the sun’s rays
can be subdivided into many wavelengths. The said classification is based on the capability of
ionizing atoms in radiation-absorbing matter. These are: a) ionizing radiation such as X-rays
and gamma rays; and b) non-ionizing radiation such as ultraviolet radiation, visible light and
infrared radiation. Solar ultraviolet radiation in the electromagnetic spectrum ranges between
100 nanometers to 400 nanometers. Ultraviolet rays are shorter than visible light but longer
than soft X-rays, which are still subdivided according to their biological effects with UV-B
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C.2 Factors that Affect Ultraviolet Radiation
Diffey (1990) gave the following as factors that affects terrestrial ultraviolet radiation
(Figure 2) : a) time of day - about 20% to 30% of total ultraviolet radiation is experienced
during midday of summer with approximately 75% occurring between 0900 hours to 1500
regions; however, seasonal variation is less when nearer the equator; c) geographical latitude
- ultraviolet radiation decreases with increasing distance from the equator; d) clouds - clouds
reduce solar radiation levels, although, the changes in the ultraviolet spectrum are negligible;
lastly, f) altitude - an increase in altitude increases the levels of ultraviolet radiation; whereas if
an area has a low altitude, e.g. sea level, there is a low level of ultraviolet radiation.
Stickler (2007), in his educational brief for the National Aeronautics and Space
Administration, enumerated the following equations for measuring the level of solar radiation.
The Inverse Square Law is a routine measure of the decrease of radiation intensity due
to an increase in distance from the source of radiation. The formula is (Formula 1):
I = E (4∏R2)
(4∏r2)
where I is the Irradiance at the Surface of the Outer Sphere; E is the Irradiance at the Surface
of the Object, which, in this case is the sun; 4∏R2 is the surface area of the object; and 4∏r2
The irradiance of an object can be obtained using the Stefan-Boltzman Law. However,
this Law is applicable only if the temperature of the object is known. The formula is (Formula
2):
E = IδT4
8
where I is the Emissivity of the Object; δ is the Stefan-Boltzman constant which is equivalent
The emissivity of an object is the factor of how well a surface can absorb and emit
energy. More precisely, it is the ration of the emitted power of the body to that of a blackbody
at the same temperature. Emissivity ranges from 0 to 1: when emissivity is equal to 1, then
the object is said to be a perfect radiator and absorber such as a blackbody. When the
Another equation which shall be of use concerns Insolation (Formula 3). Its formula is:
I = S cos Z
where I is the Insolation which is the rate at which direct solar radiation is incident upon a unit
horizontal surface at any point on or above the surface of the earth; S which is approximately
equivalent to 1000 W/m2 (this value is not absolute since it is the solar insolation during clear
days only); and Z is the Zenith Angle which is the angle from the zenith to the sun’s position.
The Zenith Angle (Formula 4) can be computed through the following equation:
where Φ is equal to the latitude; σ is the solar declination angle; and H is the hour angle
H = [15º x (Time-12)]
Sampson and Cane (1999) have concluded that intensifying levels of UV-B greatly
affects flower production: There is a considerable amount of delay of flowering, and flower
production is greatly diminished in some plants which they have tested. However, they have
observed that there was no overall effect on pollen and nectar production.
9
In Newton, Tyler and Slodki (1979), blue-green algae (cyanobacteria) was subjected to
UV-B stress. It was observed that cyanobacteria’s nitrogen-fixing enzyme system seem to be
highly sensitive to UV-B damage; and that inhibition of nitrogenase activity can take place
even other physiological functions are suppressed. Consequently, the have concluded that
the measure of acetylene reduction activity in nitrogen-fixing systems may provided a simple
up. They used various intensities of UV radiation to achieved bacteria-free algal culture. They
concluded the following: there was not bacterial growth on algal cultures inoculated on
nutrient agar for 64 hours; algal pigments were not affected by UV radiation; phycocyanin was
not inhibited; and that growth requirements were not at all altered. Overall, they concluded
that UV radiation is quite effective in obtaining and maintaining axenic algal cultures. Morton
and Derse (1968), however, observed that use of gamma radiation (as opposed to ultraviolet
radiation) on algae was effective in controlling growth where algae are unwanted. They also
recommended that a dosage of 100 krads to 150 krads is needed in a laboratory set-up.
Grad (2003) recommended that when studying aquatic organisms which are treated
with high levels of radiation, the experimentation should be subjected to frequent monitoring.
She claimed the above because recent studies have concluded that ultraviolet tolerance,
once assessed shortly after exposure, may be overestimating the true tolerance of the
organism.
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Chapter III
METHODOLOGY
A. Study Area
Field works will be conducted by the start of the summer season starting from March
ending up to July of 2008. The sites of study are beaches found in Calatagan, Batangas and
Bolinao (16º 23’ 0.83” North; 119º 53’ 87” East) is situated at the northwestern tip of
Pangasinan and the Lingayen Gulf. It is bounded on the north and west by the China Sea, on
the south by the rolling hills and plateaus of Bani, and on the east by the Kakiputan Channel.
Calatagan (13º 49’ 53.34” North; 120º 38’ 8.04” East) in Batangas comprises teh
Calatagan Peninsula between the South China Sean and Balayan Bay.
B. Algal Species
Turbinaria conoides and Padina minor are the only algae that shall be of importance to
this study because these algae are present at both sites at the same months.
Turbinaria conoides (Figure 5A) have an erect thalli, yellowish brown to dark brown in
color. It thrives mostly on sandy-corally bottom with preference to reef portions not exposed to
Padina minor (Figure 5B)is a blade flabellate, yellowish brown to whitish color, up to
seven centimeters high. They are attached to solid substrates on reef flat and upper subtidal
zone. They grown on the inner reef flats and on tidal pools on the outer portions of reef flats
(Trono, 1997).
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C. Collection of Data
This study shall observed the following abiotic factors as its parameters: pH,
temperature, salinity and dissolved oxygen content, water depth and level of solar UV-B
radiation.
thermometer will be immersed in sea water and the temperature will be recorded. The
process will be repeated thrice at a 30-minute interval. The average of the recorded
temperature on a single field work day will be the considered as the temperature of that
particular day.
The pH will be measured by employing a pH meter. The device will be immersed in sea
water and the numerical value will be recorded. The process will be repeated thrice at a 60-
minute interval.
A salinity meter will be used to record the level of salinity in sea water. The salinity
meter will be immersed in sea water and the numerical data recorded. This process will be
For the dissolved oxygen content, the Azide-Winkler method will used to gather data
(Figure 6).
Water depth will be measured using a meter stick or yard stick (or similar equipment)
Measurement of water depth will be repeated three times and the average of the data
recorded.
As for the level of solar UV-B radiation, a radiometer shall be used to record the
amount of radiation for a particular study site. This will be repeated hourly and fluctuations in
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the level of UV-B radiation recorded. The radiometer to be used should be specialized for field
works.
Field works shall be conducted from 9:00 am to 3:00 pm for 3 days on consecutive
weekly trips that will take place starting in March and ending in July. Parameters mentioned
above shall be employed. A number of algal specimens shall be collected with the
recommended minimum of 10 samples per specimen per collection day. Collected specimens
shall be placed in containers or in plastic bags (zip locks). For larger samples, these will be
placed inside containers with salt water or formalin or Bouin’s solution. Small samples shall
be placed in zip locks. However, before they are placed, a photograph must be taken before
D. Analysis of Data
Gross morphology of the algal specimens shall be observed in either the field of study
or at a laboratory. Destruction of leaf and other part of the thallus will be observed. Its length
and width will be measured using a foot rule. The surface will be categorized as rough,
smooth; shiny or slightly opaque. The intensity of coloration shall also be taken into
consideration.
and mitochondria will be observed on the two algal specimens at two different sites.
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D.3 Comparison with Control Samples
In order to establish a reliable analysis of the effects of solar UV-B radiation, a control
group will be established consisting of Turbinaria conoides and Padina minor cultivated in
vitro. These two laboratory samples shall then be subjected under ultraviolet bulbs. Gross
morphology and ultrastructure of the two mentioned algae shall be analyzed and then
D.4 Computations
The amount of radiation shall be observed using a solar radiometer. However, in order
to justify the accuracy of the numerical data produced by the solar radiometer, the Inverse
Using a statistical software package, the data gathered will be analyzed using T-test for
the comparison of the actual difference between the two means in relation to the variation in
the data. Also, One-Way Analysis of Variance shall estimate the variability between groups
compared with variability between groups (Mendenhall, Beaver & Beaver, 2006).
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Chapter IV
REFERENCES
Acra, A., N. Jurdi, A. Mu’allem, Y. Karahagopian & Z. Raffoul. Water disinfection by solar
radiation : Assessment and application. Retrieved at http://almashriq.hiof.no/lebanon/
600/610/614/solar-water/idrc.
Bischof, K., G. Peralta, G. Kräbs, W.H. van de Poll, J.L. Péres-Lloréns & A.M. Breeman.
2002. Effect of solar UV-B radiation on canopy structure of Ulva communities from
southern Spain. Journal of Experimental Botany Vol. 53, No. 379, pp. 2411-2421.
Diffey, B.L. 1990. Solar ultraviolet radiation effects on biological systems. Retrieved at
http://www.ciesin.columbia.edu/docs/001-503/001-503.html.
Essential Systems and Services for EH&S and Crisis Management. Global warming.
Retrieved at http://www.ess-home.com/news/global-warming/ozone-
depletion.asp#thumb.
Giambattista, A., B.M. Richardson & R.C. Richardson. 2007. College physics (2nd ed.).
McGraw-Hill : USA.
Grad, G. 2003. UV damage and photoreactivation : Timing and age are everything.
Retrieved at http://www.findarticles.com.
Holzinger, A. & C. Lütz. 1993. Algae and UV irradiation : Effects on ultrastructure and
related metabolic functions. Retrieved at http://cat.inist.fr/?
aModele=afficheN&cpsidt=17547275.
Mehta, R. & K. Hawxby. 1977. Use of ultraviolet radiation to achieve bacteria-free algal
culture. Proc. Okla. Acad. Sci. Vol. 57 pp. 54-60.
Mendenhall, W., R.J. Beaver & B.M. Beaver. 2006. Introduction to probability and
statistics. Thomson Brooks/Cole : Singapore.
15
Morton, S.D. & P.H. Derse. 1968. Use of gamma radiation to control algae. Wisconsin
Alumni Research Foundation. Vol. 2, No. 11 p. 1041.
Newton, J.W., D.D. Tyler & M.E. Slodki. 1979. Effect of Ultraviolet-B (280 to 320 nm)
radiation on blue-green algae (cyanobacteria), possible biological indicators of
stratospheric ozone depletion. Applied and Environmental Microbiology. Vol. 73. No.
6, pp. 1137-1141.
Stickler, G. Solar radiation and the earth system : Relating solar radiation physics to
earth & space science concept. National Aeronautics and Space Administration.
Retrieved at http://education.gsfc.nasa.gov/experimental/all98invProject.Site/Pages/
science-briefs/ed-stickler/ed-irradiance.html.
Trono, G.C. 1997. Field guide & atlas of the seaweed resources of the Philippines.
Bookmark : Makati City, Philippines.
http://asd-www.larc.nasa.gov
http://landsat.usgs.gov
http://msp.rmit.edu.au/Article_01/02.html
http://www.philippinetraveler.com
http://www.com.univ-mrs.fr/IRD/atollpol/ecorecat/images/halimac.jpg
http://www.usep.edu.ph
http://www.ecy.wa.gov
http://www.wikipedia.org
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APPENDICES
Appendix A
Figure 2. Different Types of Atmospheric Intervention Occurring when Rays of the Sun reaches the Earth’s
Atmosphere
17
Figure 3. Radiation Spectrum of Ultraviolet Rays
18
Appendix B
19
Appendix C
A B
20
Appendix D
The Azide-Winkler Method is done through the following procedure (Washington State
2. 2 ml of manganese sulfate was immediately added to the collection bottle by inserting the
calibrated pipette just below the surface of the liquid and squeezing slowly avoiding bubble
21
formation. Oxygen is introduced into the sample if the reagent is added above the sample
surface.
4. The bottle was stoppered and was made sure no bubbles were formed. The sample was
mixed by inverting the bottle several times. Again, the bottle was checked if air bubbles
were formed. If oxygen is present, a brown to orange cloud of precipitate will appear. This
is called the floc. When the floc has settled to the bottom, the sample was again mixed by
inverting the bottle several times. Again, the precipitates were allowed to settle again.
5. 2 ml of concentrated sulfuric acid was added by using a pipette held just above the surface
of the sample. The bottle was again stoppered and inverted several times in order to
dissolve the floc. At this point, the sample can be considered “fixed” and ready for storage.
Sample can be stored up to 8 hours if kept in a cool dark place. As an added precaution,
distilled water was squirted along the stopper with the cap bottle wrapped with aluminum
22