Is Feeding in Freshwater Cnidarians Dependent on Calcium?

Jennifer Goble, Kacey Hoover, Tony Kluttz, and Jeremy Proctor BIOL 2502: Lowery 10/14/2010

ABSTRACT (Replace this message with text)

INTRODUCTION Cnidarians are a phylum of aquatic organisms with specialized capturing structures called cnidocytes. Cnidarians are also diploblastic meaning they have two cell layers, the gastrodermis and the epidermis. Hydra are predatory freshwater cnidarians that are radial in their symmetry and contain tentacles that aid in the capture of food and in defense (Ozbek, Balasubramanian, & Holstein, 2009). Hydra contain special cnidocytes structures called nematocysts that contain venom that becomes released into prey when feeding. Toxins can cause paralysis of the prey.

Calcium plays an important role in Hydra feeding. Hydra get calcium from their surrounding aquatic environment. Capsules of nematocysts contain large concentrations of calcium before firing (Watson & Mariscal, 1984). The calcium concentration precipitously drops after discharge of the nematocyst. Calcium works in a second messenger reaction with cyclic AMP to help in nematocyst discharge (Rogers-Lowery, 2010). A loss of calcium could hinder the nematocysts ability to fire. EDTA and Calcium Ionophore A23187 were used in this study. EDTA binds calcium and removes calcium

Jennifer Goble from the use of hydra and Ionophore A23187 causes cells in hydra to release stored calcium (Rogers-Lowery, 2010).

Our hypothesis stated that because calcium plays such an important role in the feeding habits of cnidarians, we hypothesize that the absence of calcium will have an effect on cnidarians eating habits. This experiment looked at capturing and eating of hydra as variables to be tested.

MATERIALS AND METHODS Experimental protocol

We obtained several samples of hydra (Hydra littoralis) from Carolina Biological Supply. Regular pond water was added to three dishes determined to be controls. Calcium free pond water was then added to the nine other experimental dishes so no calcium would be present in the Petri dishes. Next we collected individual hydra and placed two of them in every dish, to increase the sample size. Both individuals were placed close together to aid in behavioral observations made using a dissection microscope. Treatment solutions consisting of 1 mL of 50 M Ionophore, 1 mL of 50 M EDTA, and 0.5 mL of 50M Ionophore and 0.5 mL of 50 M EDTA together were then added to each of the Petri dishes (three dishes for each treatment group). Hydra were the placed into each of the Petri dishes where they were allowed to acclimate to their habitat. Next, brine shrimp naupli (Atremia) were added in two drops close to the hydra to facilitate feeding. The hydra were then observed under a dissecting microscope for 10 minutes for each Petri dish. The number of prey that was captured and the number of prey that were eaten were recorded.

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Statistical analysis Because our data did not meet the assumptions of a parametric statistical test, we used a nonparametric Kruskal-Wallace to observe differences in feeding between our four groups. The KruskalWallace test is the non-parametric equivalent to an ANOVA. We compared our control to Ionophore, EDTA, and Ionophore and EDTA. Our confidence level was set at 95 percent.

RESULTS

All of our treatment groups significantly differed from one another with a p-value of 0.033 for Prey Captured and 0.022 for Prey Eaten within groups. See Table 1, Figure 1, and Figure 2 for more information.
DISCUSSION Claims Our results show that the absence of calcium does have an effect on the feeding habits of hydra. Our study looked at what would happen if calcium was excluded for use by hydra. The treatment groups all statistically differed from the control and themselves, thus enabling us to accept our hypothesis. When calcium was not present, feeding behaviors decreased, leading to loss of prey captured and eaten by hydra. Evidence Hydra showed reduced feeding behavior when exposed to calcium antagonists. Capturing and eating decreased with lack of calcium. Related Readings Our studies agreed with those of Hermansen, Arvedlund, and Fiedler (2005) found in sea anemones that are also known for their cnidocytes. Calcium antagonists helped decrease the feeding behavior seen in hydra. Calcium increases the osmotic pressure found in nematocysts (Weber, 1989). This increase allows for the release of nematocysts which also agrees with our findings. Calcium antagonists

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blocked hydra from utilizing calcium in their feeding process. Osmotic pressure was not able to build leading to less nematocysts firing and this influenced the number of prey captured which in turn affected the number of prey eaten. Reflection This study allowed us to observe the feeding habits of hydra in the absence of calcium. Although we know calcium is an important resource for nematocyst function, we wanted to see if taking calcium away would hinder the feeding of hydra. This is important when thinking of environmental impacts that may place too much or take calcium out of the freshwater hydras aquatic habitat. Because signaling methods are so advanced in hydra, it makes them a good test organism for looking at how calcium affects their signaling processes. This study confirmed that lack of calcium does affect their feeding responses.

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LITERATURE CITED

Hermansen, T, Arvedlund, M, & Feidler, C. (2005). Calcium antagonists inhibit the discharge of cnidae in response to electrical stimulation in the giant tropical sea anemone heteractis crispa ehrenberger (anthozoa). Marine and Freshwater Behavior and Physiology, 38(4), 269-274.

Ozbek, Suat, Prakash G. Balasubramanian, Thomas W. Holstein (2009). Cnidocyst structure and the bioechanics of discharge. Toxicon. 54: 1038-1045.

Rogers-Lowery, C. (2010). Biodiversity II Laboratory Manual. 65-80.

Watson, Glen M., Richard N. Mariscal (1984). Calcium Cytochemistry of Nematocyst Development in Catch Tentacles of the Sea Anemone Haliplanella luciae (Cnidaria; Anthozoa) and the Molecular Basis for Tube Inversion into the Capsule. Journal of Ultrastructure Research. 86: 202-214.

Weber, J. (1989). Nematocysts (stinging capsules of cnidaria) as donnan-potentialdominated osmotic systems. European Journal of Biochemistry, 184(2), 465-475.

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Prey Eaten Mean/SD Control 7.67 +/- 4.04 EDTA 6.00 +/- 6.928 Prey Captured Control 24.67 +/- 16.5 Ionophore 2.33 +/- 2.309 EDTA 28.67 +/- 18.9 EDTA + Ionophore 1.33 +/- 0.577 Table 1. Table shows means and standard deviations for treatment groups. Two variable were measured (Prey Eaten and Prey Captured).

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c a

Figure 1. All groups differed significatly from one another with a p-value of 0.033. Letter designations (a, b, c, and d)indicate significantly different groups.

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b a

Figure 2. Shows prey eaten by Hydra in the presense of EDTA. Hydra did not eat prey in the presense of Ionophore and Ionophore + EDTA. Experimental groups significantly differed from one another with a p-value of 0.022. Letter designations (a and b) infer statistical difference between groups.