Annona Squamosa (atis) leaves as molluscicide for Golden apple snail

Introduction Farmers, even before has so many issues in their farm and the most common problems of farmers in their farm lots are kuhols, specifically the Golden Apple Snail, which pests their plants. Golden apple snail The Golden Apple Snail is amphibious. While there is a sufficient level of dissolved oxygen in the water, they will hardly leave the water and will breathe by their gills. If the level of dissolved oxygen gets to low, the snails will go up to the surface and breathe with their lung (mostly by using its siphon, a tube that it extracts to the water surface). While filling their lung with air, the snails extract and contract their body, so that the pressure differences cause their lung to deplete and refill.

These snails have the ability to cruse away from the aquarium, so a cover glass is a necessity, as they won't survive longer than a couple of days in dry conditions. These snails have a tendency to float around the water surface, "playing dead"- but actually, they are not. The best way to tell these snails are dead is if it doesn't contract when you touch it or if the operculum (shell cover) is missing (mostly at that time, the shell will also be empty). Due to their liking of plant matter as food, most people recommend not to keep them implanted on aquariums.

Farmers of crops like “palay” who suffer from the activities of these Golden apple Snails are very unfortunate for having these pests in their farm because it was the perfect pest which can eventually damage your plant in a short period of time. Farmers’ goals when treating this problem about golden apple snail is controlling the mortality rate of the said snails not through putting ducks on their farm nor hand picking the golden apple snails, but through the use of solution which can easily kill the snails.

Golden Apple Snails live in ponds, swamps, irrigated fields, canals and water-logged areas, can even survive in polluted water with low oxygen levels, bury themselves in moist soil during the dry season, lie dormant for 6 months and become active again when the soil is flooded. They stay submerged during the day, usually hiding in dense vegetation near the edges or the surface. They are more active during the night, often leaving the water in search of new feeding areas. They can live for 2 to 6 years.

Adults mate any time of the day, usually among crowded plants where there is continuous water supply throughout the year. (http://www.kelabit.net/news/special_report_01.html). Molluscicide is a solution, chemical or organic which can kill mollusks. In this study the researchers will use the organic molluscicide as killer for Golden Apple Snail, and the main ingredient will be Annonasquamosa (Atis) seed.

Annonasquamosa is a small (2-4"), knobby fruit with soft, creamy white flesh often having a minty or custardy flavor. The sugar apple is extremely popular throughout the tropics, especially in climates where the cherimoya can not be grown. (http://www.tradewindsfruit.com/sugar_apple.htm). Sugar apple, believes to be native from the Caribbean region and the northern South America but is now cultivated in most of the tropical countries throughout the world.

This fruit is known by its numerous vernacular names, mostly by the languages of the different countries. Some countries loosely apply the name, custard apple to this fruit, which is misleading and technically incorrect. There is another fruit in the same family, which looks similar but not as common, known by the actual name of custard apple (annonareticulata). So don't be confused by the same name as both fruits are different. Sugar apple, as the name says it all, is sweet as sugar. The flesh nearest to the rind, tastes like sugar crumbs too. It is usually conical in shape but sometimes, it may be almost round. It is easy to tell when it is ready to eat.

The rind is thick with knobby segments but will turn soft and crack open, releasing a sweet aroma when it rips. (http://www.vegtalk.org/fruits/sugar-apple-annona-squamosa-t3594.html#p9153). Since the researchers found out that Atis seed has its toxicity level enough for being a component fro killing the Golden apple snail and since it is also organic, the researchers have decided to make atis seed extract as molluscicide. Statement of the Problem

A study will be conducted to test the effectiveness of Annona squamosa as molluscicide for Golden Kuhol. Specifically, it aims to answer the following questions: 1. Is the Annona squamosa Molluscicide effective in terms of: a. Mortality rate

b. Duration of Potency 2. Is there a significant difference in the effectiveness of the Annona squamosa molluscides under different concentration (50%, 75%, and 100%) a. Mortality rate b. Duration of Potency 3. Is there a significant difference in the effectiveness between the Annona squamosa Molluscicide and the commercial molluscicide in terms of: a. Mortality rate b. Duration of Potency

Significance of the Study This research study will benefit the following sector: Plant Physiologists. Plant Physiologists are the people who study about the function of a certain plant. This study will help this group of people to have more information about the other uses of Annonasquamosa, for example, as a molluscicide. Farmers. This study can help farmers in having affordable and effective molluscicide against pests on their farm lots.

Department of Agriculture. Instead of letting Atis seed go to waste these can be made useful by making it into an effective molluscicide. They can use Atis as one of their products in agriculture and make it known into other places. Growers of Atis. This study will help tose who are planting Atis to have new source of income.

The Society. It will make them aware of the availability of the natural organic insecticide that is less expensive yet safe and effective. Future Researchers. It will serve as a reference material for their future research. Scope and Delimitation

This study aims to control the mortality rate of Golden Apple Snail by using the extract of Annona squamosa(Atis) seed. The Golden apple snails which were usually present on farms are involve in this study, because those snails would be the tester on the effectiveness of the Annona squamosa seed extract Molluscicide.

This study will be conducted in Naga City Science High School for the school year 2012- 2013 and the researchers as the observers. The aspects looked into where this research would be possible for control for the said pest, the problem and the said methods for treating them. The study on Annona squamosa seed as molluscicide for Golden Apple Snail limits on the effectiveness of the extract of the seed. It does not include the appearance of the mixture. Definition of Terms

Molluscicide. It is a solution that can kill molluscs. Atis seeds. It will be the main ingredient of the AtisMolluscicide. It has toxic that can kill the Golden Apple Snail. Mortality. This refers to the death rate of the Golden Apple Snail. Potency. It determines the effectiveness of the Atis Molluscicide. Research Paradigm

Figure1. Research Paradigm Chapter II REVIEW OF RELATED LITERATURES AND RELATED STUDIES Related Literature Golden Kuhols came from South America. It was introduced in Taiwan as a food. It became an aquarium pet and helps in cleaning tanks. It multiplied rapidly until it reaches the waterways. (2008) (http://juanmagsasaka.blogspot.com/2008/08/protect-rice-seedlings-against-golden.html) It was first introduced in Asia in the year 1980 with expectations that it can be a high-protein food source. However, it has invaded the rice fields in Asia. The young rice seedlings were eaten by these snails. (http://www.jstor.org/discover/10.2307/4314515?uid=3738824&uid=2&uid=4&sid=21100870152841) Annona squamosa or known as sugar apple is the most widely grown of all the species of Annona. When eating this fruit, the seeds are always left out due to its toxicity. (http://www.ehow.com/facts_6947915_sugar-apple-fruit.html).

The powdered seeds as well as the pounded leaves are used as insecticide in India. The heat-extracted oil from the seeds can be used against agricultural pests. (http://www.oocities.org/athens/academy/4059/atis.html). It can also be used as a lice remover; however, it must not reach the eyes because it can cause blindness. If it is applied to the uterus, it can cause abortion. (http://www.hort.purdue.edu/newcrop/morton/sugar_apple.html#Toxicity)

The seeds of A. squamosa contain annonaceous acetogenins which have cytotoxic activities. (http://www.globinmed.com/index.php?option=com_content&view=article&id=83499:annona-squamosa&catid=703:a). It is said in one study that the acetogenins from the seed are more toxic than the synthetic pesticide. (http://www.stuartxchange.org/Atis.html)

Molluscicides are used to control slugs and snails. Slugs and snails belong to the mollusks group. Metaldehyde is one of the most commonly used active ingredients in molluscicides. Molluscicides can be manufactured as a colored pellet or a spray. Since molluscicides are commonly chemicals, there are other ways to control mollusks. This includes using traps or barriers. There are slug and snail baits that contain iron phosphate as the active ingredient. Iron phosphate is less toxic than metaldehyde. (http://toxipedia.org/display/toxipedia/Biological+Properties+of+Molluscicides).

Copper Sulfate can be used as molluscicide. It has been used for about 40 years now. It is still used as molluscicide in some areas. Although it is effective, it still has limitations; it is more effective in other habitats. (http://whqlibdoc.who.int/monograph/WHO_MONO_50_(part4).pdf) Currently, only copper sulfate and copper chelate formulations have been utilized as a viable molluscicide treatment in open water systems with proper permitting. Niclosamide may be used for controlling snail (mollusks) populations in aquaculture ponds. (http://www.glmris.anl.gov/documents/docs/anscontrol/Molluscicides.pdf)

According to Dr. Eduardo Quisumbing, the seed area is considered a powerful irritant to the conjunctiva; the roots are considered a drastic purgative. So considering the parts of the articles stated above it would be a great help in proving the researcher’s study. (http://www.pinoyecofarmer.com/2010/10/21/atis-leaves-developed-antidengue-so lution/) Related Studies

Study shows that Annona squamosa plant can be used as a molluscicide. Among the seeds, bark and the leaves of the plant, it was observed that the highest molluscicidal activity of the Annona squamosa plant is in the seed extracts. The acetogenins extracted from the seeds of the plant were more toxic than synthetic pesticicides.

The combination of the powdered seed of A. squamosa, the oil from the cedar, Cedrus deodara Roxh., and from neem, Azadirachta indica A. Juss., have more toxic than the separate constituents of the cited plants. (Singh & Singh , 2001) Another study was conducted about the effects of the A. squamosa Linn. and Lawsonia inermis Linn. (Henna) and their combinations with other herbal molluscicides, and acetogenins extracted from the seeds of A. squamosa Linn., on the reproduction of Lymnaea acuminata.

It was observed that the plant- derived molluscicides affected the mortality rate of the snails. However, when the snails were put back to the freshwater, it regained its capability to reproduct. (Singh and Singh, 2004) A study was performed, by other proponents, to know the molluscicidal properties of some species of Annona. There were six species used in this study and were observed if they have the capability to cause death of Biomphalaria glabrata. Biomphalaria glabrata is a tropical freshwater snail indigenous to ponds, marshes and streams. (http://biology.unm.edu/biology/esloker/pi/WhatIsBiomphalaria_glabrata.pdf)

The results of the experiments showed that the majority of the extracts of the Annona species have the properties to harm the Biomphlaria glabrata, moreover, the Annona muricata L. (leaf) and Annona squamosa Veil. (root) was given emphasis on because of their additional toxicity towards the snail eggs. (Dos Santos AF, Sant'Ana AE., 2001) Other fruits were tested to know if they have a molluscicidal activity. A combination of jackfruit, durian and breadnut peels was studied to control Golden Kuhol. The researcher used the rotten peels of the said fruits in making the alternative molluscicide. The study has 5 treatments and each treatment composes of different masses of the different fruits.

The researcher added 45 g and 10 g to the fourth and fifth trial respectively. It is stated in the study that the jackfruit peel have a foul smell that can kill the Golden Kuhol. (Villa Agustin, 2012) (http://www.scribd.com/emmanuel_agustin_2/d/79325542-The-effect-of-jackfruit- durian-and-breadnut-peels-can-control-golden-apple-snail#) Barringtonia racemosa fruit and seed extracts were experimented to know if it has a molluscicidal, cercariacidal, larvicidal, and antiplasmodial properties. The extracts of the stem bark is said to be toxic to mosquitoes, snails and brine- shrimp. The World Health Organization’s method for examining molluscicides was used in this study. Biomphalaria pfeifferi snails were collected from a stream in Edun-Abon in Nigeria.

The snails were separately exposed in the concentrations of the B. racemosa fruit and seed aqueous extracts. Niclosamide is a chemical molluscicide that also used in this experiment as the control molluscicide. There were 10 snails used per extract or niclosamide dose and was exposed within 24 hours. It has three replicates. After that, the observations were recorded. The fruit extract of B. racemosa has stronger molluscicidal activity than the sed extract of the plant. (http://redalyc.uaemex.mx/pdf/856/85630505.pdf) Chaper III

METHODOLOGY Research Method This study will use the experimental method of research. In this research method, the researchers will manipulate the variables involve in the study. The purpose of this method is to determine the effectiveness of Annona squamosaas Molluscicide for Golden Kuhol. Set Up

There will be 4 set ups: for 50% concentration, 75% concentration, 100 % concentration of Annona squamosa (atis) molluscicide, and for commercial molluscicide. In each set up, there will be 3 kuhols. Then, the researchers will conduct an observation about the effect of the product and its difference to the commercial molluscicide. Materials Needed

In making the Annona squamosa (atis) as molluscicide for golden kuhol the following materials are needed: Annonasquamosa (Atis)seeds Distilled water Spray bottles Strainer/ Cloth\ Copper sulfate Mortar and Pestle