ABSTRACT

Microorganisms are too small to be seen with the unaided eye, therefore, they must be observed with a microscope. Due to the fast innovations and advances in microscopy, we are now capable of examining, determining and observing them. This experiment, which utilized the hay infusion as the residing area of microorganisms, investigated the living organism in a drop of water, their manifestation of the different attributes in life. During the run of the experiment, paramecium and specie 1 were observed and were tested to determine the impact of different stimulus to microorganisms in three setups. At the end of the experiment, the variety of setups has been shown to affect the number and kinds of species seen, such as placing hay infusion in dark and the other one, exposed to sunlight, adding brine and sugar solution on the setups. The third section of this paper will deal with relative number of species observed and seen. This paper tackles how microorganism thrive and exemplify mechanisms for survival in the different setups for comparison Moreover, this proves that even in a small drop of water, still, life can exist and microorganisms prosper.

I. INTRODUCTION

Life on Earth was said to have begun 3.5 billions of years ago. Until now it is still dominating, counting and adding years to life’s age. For an organism to be considered living or possesses life, they must portray different attributes of life. This include mainly by movement, irritability, contractility, perpetuation of life, adaptation, nutrition and growth and development. Living organisms come across our vision because of their relative size. We easily see living organisms that are huge in size like gorillas and for some cases, even small organisms like ants. However, not all living organisms are of size that we can easily visualize. These organisms, the microorganisms, are very minute and microscopic and needs special instruments like the microscope in order to cross periphery of our vision.
Microorganisms, from the Greek word “mikros” meaning small, and “organismos”, which means organism, also spelled micro-organism, micro organism, microorganism or microbe is a microscopic organism that comprises either a single-cell(unicellular) or cell clusters. These include bacteria, fungi, archaea or protists, but not viruses and prions, which are generally classified as non-living. Microorganisms live almost everywhere on Earth where there is liquid water, including hot springs, on the ocean floor, and deep inside rocks within earth’s crust.
Under the microscope, the microorganism that we mostly found and easily noticed in this research were the paramecia of the Phylum Protozoa and specie 1 due to its observable movement and features.
Protozoa means “first-animal” which generally describes its animal-like nutrition. All protozoa live in areas with large supplies of water. However, Volk mentioned that even in the soil, protozoans live. They are unicellular, eukaryotic chemo heterotrophic organisms. Prescott defined Protozoa as Protists exhibiting heterotrophic nutrition and various types of locomotion. They occupy a vast array of habitats and niches and have organelles similar to those found in other eukaryotic cells and also specialized organelles.
Under this Phylum, is the free-living, non-pathogenic Paramecium which belongs to class Ciliata. Paramecium is a small one celled(unicellular) living organism that can move, digest food, and reproduce. They belong to the kingdom of Protista , which is a group(family) of similar living microorganisms. Microorganism means they are a very small living cell. The paramecia are known for their avoidance behavior, if it encounters a negative stimulus, it is capable of rotating up to 360 degrees to find an escape route. In addition to paramecium, the specie 1, that has a slender body structure and moves in a spiral-like motion, is another type of specie that is similar to a worm and always in close proximity to the debris but eventually leaves them .
In this paper, these organisms will likely to be discussed and will be tackled more as these organisms undergo several phases of concentration and analysis during the experiment.

Scope and Limitation of the Study

Due to the limitation of laboratory materials and time on this experiment, the incubation of the hay infusion had the run time of only five days and the organisms were observed for no more than six hours. This is probably the reason why there were limited species observed under the microscope namely the paramecium and specie 1. In addition, we are not able to identify all microorganisms on the different slide because of the limited knowledge and data about microbes.

II. MATERIALS AND METHODS

This study used cotton, dropper, glass slide, cover slip, microscope, brine solution, sugar solution and hay infusion.
In making hay infusion, any natural water will work except tap water. Tap water that we usually use is treated with chlorine, enough to remove and kill microbes and to make sure that what we drink is potable. Then the water is placed in container, preferably the ones that has already impurities, as it is going to help increase growth of microorganisms in water. Hay or grass, and dried leaves are added to the container with water. Hay infusion should be placed at a room temperature or above for several days. Add water as it evaporates. After 5-10 days, the water should turn dark and turbid; monitoring and examining a drop of water will reveal a large number of organisms.
In performing this experiment our professor required us to make different set ups of hay infusion, one that is exposed to light, the other is placed in a dark corner and a hay infusion that utilized a stagnant water in a plant. The first procedure is to lay two to three strands of cotton on a microscope slide and apply a drop of water from the hay infusion samples. Using the microscope, the drop of water was observed under LPO and HPO. We also placed a drop of salt solution (salt and water) and sugar solution (sugar and water) after, at the right edge of the glass slide. The procedure will be done again for the other hay infusion samples.

III. RESULTS AND DISCUSSIONS

The three setups were prepared for the evaluation of microorganisms. These were the hay infusion with a stagnant water from a plant(A), second, hay infusion exposed to sunlight(B) and hay infusion with a pond water placed in the dark(C). Each hay infusion was incubated for
Findings shows that there were more organisms in the preview of setup A, the hay infusion with stagnant water from a plant, namely paramecium and specie 1. The paramecia, in the figure 1 below, under the LPO(10x) of the microscope, are transparent like water, some were round and others were elliptical in shape and very small. They go near the debris or non-living objects through gliding on the water sample , sometimes bump to other paramecium, then moves away from the field of vision at a very fast pace. When shifted to HPO, the paramecia was seen larger and leaves the field of vision at once. Just like what it does that was seen in the LPO, it slides along with other paramecium.
The specie 1 was also observed from a drop sample of stagnant water under the microscope. Because of the narrow familiarity about microorganisms, we are not able to identify and name the specie. In the figure 1 below, specie 1 was slender and slim in structure, and brownish red in color. Under the LPO (10x), assuming that the LPO was like our normal vision, the specie 1 was no more than 1 inch in measure. It also go near the impurities on the slide then also move away by its twisting its whole body like a spiral. Under the HPO, the specie 1 was viewed in a bigger manner. The specie 1 was still, coils itself to move on the slide, it has lines on its body and we can say that it has similar characteristics like a worm. The paramecium and the specie 1 ignores each other and tend to keep away from each other.
To test their response to sudden changes in their environment, a drop of sugar solution was placed on the slide (see figure 2). The paramecium, instantly compact themselves together and the specie 1 was twisting and coiling itself repeatedly. After a few seconds the paramecium stopped moving and so with the specie 1; they just go with the flow on the water on the slide then immediately disappear on the field of vision. Another solution was placed, the brine solution (see figure 3). After placing a drop on the slide, the paramecium was jumble in movement, as if it wants to escape out of the water but does not know how to. When the drop of salt solution was placed, the paramecium spread themselves, unlike to the sugar solution that made the species compact. After a few seconds, the paramecium disappeared. The species 1, likewise, moved in a disarray movement, the second after, it also became immobile.

LPO(10x) HPO(40x)
Fig.1. Paramecium and Specie 1 of hay infusion with stagnant water from a plant.

LPO(10x) HPO(40x)

Fig.2. Paramecium and Specie 1 with sugar solution.

LPO(10x) HPO(40x)

Fig.3. Paramecium and Specie 1 with brine solution.

There were also paramecia in setups B and C, hay infusion exposed to sunlight and hay infusion that were kept in the dark respectively. However, there were less paramecium in the hay infusion on the dark than the hay infusion placed in the sunlight. This was maybe because of the absence of sunlight, which hindered photosynthetic materials to make their food. Therefore there were least food for the Paramecium. However, there were still Paramecium observed because of the bacterial growth in the hay infusion that served as the food for the organism. When the brine solution was laid on the slides of set ups B and C, just like what happened on the paramecium on the hay infusion that used stagnant water, the paramecium moved in a random movement at a rapid pace. The paramecium in both setups were eager to escape but failed because the whole slide were concentrated with salt. The paramecium were no longer moving and disappeared after the few seconds of exposure to salt. When sugar solution was placed on the setups B and C, the paramecia compact themselves with one another, then, after a few seconds the paramecia were stationary, then disappear.

LPO(10x) HPO(40x)
Fig.4. Paramecia in dark hay infusion.

LPO(10x) HPO(40x)
Fig.5. Paramecia in hay infusion exposed to light.

IV. SUMMARY AND CONCLUSION

The Paramecium and Specie 1 were found to prosper in a wet environment which in this experiment, is the hay infusion. The location of the hay infusion affected the growth and accumulation of organisms observed. The concentrations that were utilized to test their irritability and response to a certain stimuli, namely the brine solution and sugar solution, likewise affected the length of time the microorganisms lived and from that situation, we can say that the paramecium and specie 1 have poor ways to adapt to sudden changes on their environment, because they died eventually after exposing them to the different concentrations. The number of days the hay infusion was incubated also have an effect on the number and kinds of microorganism viewed. Hence, for successful growth of other organisms, the hay infusion should be reared under optimum temperature and must be nurtured ten days or more. It was clear indeed that in this experiment, a small portion of water could be the habitat of living organisms which we do not actually see, but exists there .