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Food Preference by Fruit Fly

Influenced By Yeast Content

Principle Investigator:

Melvin Elias

Undergraduate Student

Biology

Tennessee Tech University

Tennessee Technological University Cookeville, TN 38505

Biological Communications (3920)

April 15, 2010

Table of Contents

Executive Summary................................................................................pg. 2

Aims of the Proposed Study            …………………………………………...pg. 3-4

Keywords………………………………………………………………..pg. 4

Introduction…………………………………………………………… .pg. 4-7

Materials and Methods……………………………………………….. .pg. 7-9

Experiment Design...................................................................................pg. 9-10

Project Time Line……………………………………………………….Pg. 11

Data Form……………………………………………………………….Pg. 12-13

Expected Results and Benefits………………………………………....pg. 13

 Personnel and capability statement of Investigators………………...pg. 14

Ethic Statement…………………………………………………………pg. 14

Disseminating Research Results………………………………………..pg. 15

Literature Cited…………………………………………………………pg. 16

Budget…………………………………………………………………….pg. 17

                        Executive Summary

Fruit Flies, Particularly Drosophila melanogaster are, small, about 4-8 mm in length

including the wings (fruit flies used for this specific experiment will contain vestigial wings to

prevent them from flying away). Fruit flies can be identified on its bright red eyes. They can

be found throughout the world, in homes, food processing plants, warehouses, grocery stores,

wineries, restaurants and other structures.

While reducing the caloric intake by diluting foods containing yeast, such diet reduce aging, but in the same vein, this caloric restriction reduces the availability of yeast in the diet thus shortening the life span due to starvation. Many factors change food preference of both female and male larvae. In D. melanogaster larvae, research have been shown that, a clear preference exists for the highest yeast concentration in food. Conversely, in larvae of the same species lacking a gene known as GS1189, larvae did not show a preference of various types of foods including yeast.

Interestingly, ripe coffee fruit has the potential to attract fruit flies to oviposit. This ripe coffee fruit (which was found to contain a high content of alcohol, in turn, suggesting the existence of yeast) is more attractive to female fruit flies than males. What’s more, the smell of recently crushed ripe coffee fruit is more attractive than that of intact ripe fruit and unripe fruit. Temperature is another factor, which can help fruit flies in their development. 

AIMS OF THE PROPOSED STUDY

Recent  literature research has fueled and at the same time led me to explore a more clean environment in which can be eventually achieved in third world countries constantly battling with the issue of an overwhelming population of D. melanogaster. By Knowing the biology, nutrient requirements, and other aspects of D. melanogaster it is possible for third world countries to rid of these undesirable insects by using fly traps containing yeast content to attract this insects thus reducing fruit fly population and eventually extinct such organism which can also be troublesome to farmers. I plan to accomplish this research by setting up an experimental design in which will contain the following. I will set up a total of twelve (for each run) 3.785-liter mason-like glass jars with four bottle caps in it each jar. Each bottle cap will contain a different type of food (apple, pineapple, banana, and control media). In each jar, only subjects from the same the vial will be introduced. After fifty-five minutes, data will be collected (using a data form sheet) by counting the subjects in each bottle cap containing food to determine if in fact preference for yeast does exist. The experiment will be repeated three times, in which each run will contain new flies.

The purpose of this experiment is to measure food preference selection (influenced by yeast concentration) by counting the subjects on each food source with different content (banana, apple, pineapple, and standard media) by Drosophila melanogaster. My null hypothesis is that there will be no difference in food selection containing yeast by D. melanogaster.

Keywords—Selection, fruit fly, characteristics of nectar, Gender, media reception, genes,

potential, behavior, Drosophila melanogaster.

INTRODUCTION

            Fruit flies specifically Drosophila melanogaster are small about 4-8 mm in length including the wings (fruit flies used for this specific experiment will contain vestigial wings for preventing them from flying away). Fruit flies can be identified on its bright red eyes. The head and thorax are tan in color with the abdomen somewhat darker (males have a larger dark abdomen than females). These insects can be found throughout the world, in homes, food processing plants, warehouses, grocery stores, wineries, restaurants and other structures.

Populations tend to be greatest in late summer and early fall as they infest fruits during the harvest season. D. melanogaster are generally found hovering around decaying vegetation and overripe fruit. Fermenting materials, such as leftover beer or soft drinks, also are a favorite food of these flies. It was found that most insects if not all need yeast in their diet for growth and development with mosquitoes being a little more demanding than D. melanogaster. Furthermore, larvae are assumed the same way as adult fruit flies in that no different diet requirement is needed to survive; interestingly vitamin B₆ and cholesterol are essential for D. melanogaster, but without yeast, fruit flies will not develop or grow even in the presence of the essential minerals and cholesterol (Tatum 1939).

Yeast is very important to fruit flies because it contains an important factor, which can be adsorbed and inactivated by hydrolysis. Furthermore, this phenomenon accelerates the growth of D. melanogaster (Tatum 1939). In other studies, it has been shown that the amount of yeast concentration in diet is directly connected with aging in D. melanogaster (also known as the caloric intake effect). While reducing the caloric intake by diluting foods containing yeast, such diet reduces aging, but in the same vein, this caloric restriction reduces the availability of yeast in the diet therefore shortening the life span of D. melanogaster due to starvation (Magwere et al 2004).

Researchers now know that insects must be able to evaluate their nutrient requirements as well as the nutrient supply offered by the environment, and translate the resulting information into appropriate behavioral responses. In fact, it has been noted that D. melanogaster change their food preferences depending on their physiological or environmental conditions. If they cannot change their food preferences to coordinate energy states of their body with food availabilities, then it must difficult for them to survive the shortage or quality change of their foods. Many factors change food preference of both female and male larvae. For example, during the development of an organism, an apparent change can be observed from high protein/low lipid diet towards low protein/high lipid diet. In addition, the fruit fly has a range of feeding characteristics that can be broken for behavioral analysis which can provide a genetically accessible system to study the molecular mechanisms which organize feeding behavior with sensory signals.

In D. melanogaster larvae it has been showed that a clear preference exists for the highest yeast concentration in food. Conversely, in larvae of the same species lacking a gene known as GS1189, larvae did not show a preference of various types of foods including yeast (Ryuda et al. 2008). In the research by Ryuda et al. (2008), When larvae were introduced to the center of the two wells (one filled with yeast-containing agar and the other well just containing agar) the larvae lacking the gene GS1189 took significantly longer to reach the yeast containing agar than the controlled group. These results indicated that the mutant larvae were defective in their chemosensory system. In fact, results suggested the possibility that the GS1189 larvae were damaged in their olfactory and gustatory systems (Ryuda et al. 2008).

Interestingly, ripe coffee fruit has the potential to attract fruit flies to oviposit. This ripe coffee fruit (which was found to contain a high content of alcohol in turn suggesting the presence of yeast) is more attractive to female fruit flies than males. The odor of coffee produced a 2.5-fold increase in the number of female landings on fruit models for oviposition. In addition, it was shown that the odor of intact or crushed coffee fruit is more attractive to both sexes of the fruit flies compared to lower-ranking hosts and nonhosts when exposed to laboratory wind-tunnel and field conditions. The smell of recently crushed ripe coffee fruit is more attractive than that of intact ripe fruit and unripe fruit. In addition, females carrying a greater egg load are more attracted to the smell of ripe coffee fruit than are immature or virgin females. This suggests that females carrying a large egg load are more likely to spend a greater amount of time on the ripe coffee fruit and at the same time attracting males to the site due to pheromones production by the female fruit fly (Warthen et al. 1997). Temperature can also help fruit flies. When nutrient availability is low, simultaneous adaptation to a cold environment leads to a more efficient allocation of resources to adult size. Additionally, it has been previously suggested that a large adult body size enhances fitness at low temperatures only telling that when nutrient availability is low simultaneous adaptation to a warm environment leads to increasing larval survival by reducing the feeding associated costs. For natural populations it seems both in moderate and tropical regions, D. melanogaster face low nutrient availability and evolve to the opposite ends of the adult size versus larval survival trade-off (Bochdanovits and De Jong 2003). The hypothesis of this research is that all D. melanogaster will prefer a sample of fermenting banana to other fruits (apple, and pineapple) and to standard media culture. The objective of this research is to determine the amount of subjects (D. melanogaster) preferring yeast containing foods.

MATERIALS AND METHODS

            Vestigial wing D. melanogaster were collected in Cookeville, Tennessee during the fall semester of 2010. From this large stock many generations were created using a set up of 24 vials for each generation. A vial has a diameter of 2.2 cm and 6.3 cm in length. The standard media used for this experiment known as formula 4-24 contains anti-mold inhibitor manufactured by Carolina Biological Supply Company provided by the genetics department. The culture media will be prepared using a 100 ml container. The prepared culture mixture will contained a 1:1 ratio. Each vial will contained 10 ml of prepared standard media (no yeast added except for a very minute amount added by the manufacturer). Forty-eight bottle caps from 29.57-ml water bottles will be used. Each bottle cap will contain pineapple, fermenting banana, apple and standard media respectively. These bottle caps will be placed inside twelve 3.785-liter mason jar-like glass container with diameter opening of 11.43 cm (4.5 inches). A micromesh sunscreen made of nylon will be used as a lid to allow airflow into the glass container and at the same time keeping fruit flies from escaping. 50 ml of ether will be used as a fruit fly morgue to avoid

further reproduction of used fruit flies. A data form (labeled Table 2) will be used to collect data after fifty-five minutes of introducing D. melanogaster into the glass jar (Hardy 2004).

Scientific Method—Table 1 employed for observation

Experimental Design—Table A

Experimental Design—Table B

Experimental Design—Table C

Project Timeline

Key

Data Form—Table 2

Biology Department

Drosophila melanogaster Treatment plan monitoring

Location: __________________________________________________________________

                    (Facility name)                                 (City)                (County)          (State)   

Sample Taken by: ___________________________________________________________

                                (Name)                     (Position)                    (Organization)

Run1

Run 1

Run 2

Run 2

Biology Department

Drosophila melanogaster Treatment plan monitoring

Location: __________________________________________________________________

                    (Facility name)                                 (City)                (County)          (State)   

Sample Taken by: ___________________________________________________________

                                (Name)                     (Position)                    (Organization)

Run 3

Run 3

EXPECTED RESULTS AND BENFITS

            I expect to reject my null hypothesis. I will be able to manipulate the amount of time of banana to be fermented this will ensure a greater amount of yeast presence in the banana. On the other side of the coin, these expected results will be available to use as trap flies reducing the population of D. melanogaster in very inexpensive manner benefitting farmers and perhaps the rest of the human population. Results are expected to agree with literature source identified for this research. These methods then can be used in third world countries where scarce pest control exists.

PERSONNEL AND CAPABITLITY STATEMENT OF INVESTIGATORS

Faculty Research Advisor: Dr. Daniel Combs.

Bioethics Statement

• Conduct research in way that is consistent and accepted by scientific and teaching methods.

• Maintaining honesty and integrity in all professional endeavors.

• Comply with all laws and regulations that apply to the treatment of study disease and other aspects of professional conduct.

• Be respectful in professional interactions, avoiding discrimination, based on race, gender, sexual orientation, religion, or age. Treat colleagues, students, and employees fairly.

• Be constructive and professional in evaluating the work of colleagues, students, and employees.

• Provide recognition of past and present contributions of others to science, and present one’s professional opinions only on those topics for which one has training and knowledge.

• Promote the free and open exchange of information, not withholding information to substantiate a personal or scientific point of view.

• Be candid about potential conflicts of interest in the conduct of professional duties.

DISSEMINATING RESEARCH RESULTS

The Results of my research will be disseminated in many ways.  I plan to present my results in class using a power point presentation in which tables, figures, and other adequate images pertaining to my research will be included.  I will also publish a website in which I will be posting ongoing results of my research. Finally, I will write a final detailed manuscript along with a poster for final presentation of my research that will also include figures and tables.

Literature Cited

1. Bochdanovits, Zoltan., and G. De Jong. 2003.Experimental evolution in Drosophila melagonaster: interaction of temperature and food quality selection regimes. Evolution. 57:1829-1836.

2. Gomulski, Ludvik. M., G. Dimopoulos., Z. Xi., M. B. Soares., M. F. Bonaldo., A. R. Malacrida.,and G. Gasperi.,2008. Gene discovery in an invasive tephritid model pest species, the Mediterranean fruit fly, Ceratitis capitata. BMC Genomics. 9:243.

3. Hardy, J. K. (2005). Copyright: Chemometrics. http://ull.chemistry.uakron.edu/chemometrics/index.html (08 April 2010).

4. Magwere, Tapiwanashe., T.Chapman., and L. Partridge. 2004. Sex differences in the effect of dietary restriction on life span and mortality rates in female and male Drosophila melangaster.Gerentology 59:3-9.

5. Ryuda, Masasuke., S.Tojo., T. Tanimura., S. Tsuzuki., and Y. Hayakawa. 2008. A gene involved inthe food preference of larval Drosophila melanogaster. Insect physiology 54:1440-1445.

6. Warthen, J. D., E. B. Jang., J. Lee., D. R. Lance., and C. D. O. McInnis.,. 1997. Volatile, potential Attractants from ripe coffee fruit for female Mediterranean fruit fly. Chemical Ecology. 23:1891-1900.

7. Tatum, E.L. 1939. Nutritional Requirements of Drosophila melanogaster. Proc Natl Acad Sci U S A. 25:490-497.

BUDGET