Amirul Proposal

8/4/2019 Amirul Proposal

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1.2 STATEMENT OF PROBLEM

It is known that GAS is pest for the rice ( Sison J.A 1985) . GAS bring a lot of problem to

farmers not only Malaysia but nowadays globally. Farmers cultured the snail indoors but when

market response was poor many snail projects were abandoned and in many instances the snail

escaped and ravaged the rice crop with losses running into millions of dollar (Naylor, 1996).

Golden Apple Snail (GAS) (Pomacea spp) is considered a major pest in the rice ecosystems of

Laos, especially in lowland rice fields (Douangbupha et al, 1998). According to Philippines Rice

Research Institute GAS bring to agricultural economic losses $ 1 billion losses to Philippine rice

crops in 1980s and 55-248 billion/year (globally). In Malaysia GAS spread rapidly following its

occurrence in Keningau in 1992. To date the districts infested with the pest include Keningau,

Tenom, Sook, Nabawan, Tambunan, Papar, Penampang, Kota Kinabalu, Beaufort, Kuala Penyu,

Tuaran, Sipitang, Kota Belud and Kota Marudu covering an area of about 5,000 ha (Teo et al.

2002).

The protein source of choice for most fish and crustaceans is fishmeal or 'trash fish'

(small fish forming the low-value component of commercial catches). However, supplies are

declining and prices are increasing. There is also an increasing conflict between use of trash

fish/fishmeal for aquaculture, or for human consumption (particularly for low-income indigenous

people). One reason for insufficient supply of trash fish for fish meal manufacture is because

about one third of the fishery catch is thrown overboard. Fishers need improved technology so a

higher percentage of the catch can be landed (Peter Edwards et al. 2004). There is growing

awareness of the danger and unpleasantness of air and water pollution, and it is likely that these


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issues will continue to be of great concern to the fish meal industry (M. L. WINDSOR 2001).

Every year price of fish pellet was increase because increasing price of raw material. Rising

global demand for fishs trash has increased the pressures on wild fisheries with catches no

longer able to satisfy demand. This is resulting in declining catch numbers and more juvenile fish

being removed before they have bred.


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1.3 SIGNIFICANCE OF STUDY

These studies will perhaps a foundation of resolving matter for help paddy planter and

aquaculturist. This is due to the decreasing attack of GAS will be reduce by increasing demand

and gives them side income for selling the GAS from their paddy farm. Aquaculturist also get

benefit because their can get fish pellet with a low price. This is because decreasing of raw

material price in using GAS meal in fish pellet production.


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1.4 OBJECTIVE

The aim of this study is to determine the potential of GAS as protein sources for fish

pellet production. This project will be dividing into two phase: first phase will be focuses in

determination of moisture and dry matter, ash, crude lipid, crude protein, crude fiber. Second

phase will stress in determination of perfect ratio in fish pellet production .This experiment

would done to Pangasius sutchi fingerling to determine Growth performance, Feed Conversion

Ratio (FCR) ,and food efficiency of all fingerling to determine the best ratio of formulated pellet.

To determine the nutritional composition in Golden apple snail powder. To determine the optimum proportion of a mixture of Golden apple snails powder in fish

pellet.

To determine effect of Golden apple snail powder in Pangasius sutchi fingerling.


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CHAPTER 2

LITERATURE REVIEW

2.1 INTRODUCTION TO Pomacea canaliculata.

2.1.1 TAXONOMY

Taxonomy ofPomacea canaliculata(Lamarck, 1819) is below:

Kingdom : Animalia

Phylum : Mollusca

Class : Gastropoda

Order : Caenogastropoda

Superfamily : Ampullarioidea

Family : Ampullariidae

Genus: Pomacea canaliculata(Lamarck, 1819)
http://en.wikipedia.org/wiki/Jean-Baptiste_Lamarckhttp://en.wikipedia.org/wiki/Jean-Baptiste_Lamarckhttp://en.wikipedia.org/wiki/Jean-Baptiste_Lamarckhttp://en.wikipedia.org/wiki/Jean-Baptiste_Lamarckhttp://en.wikipedia.org/wiki/Jean-Baptiste_Lamarckhttp://en.wikipedia.org/wiki/Jean-Baptiste_Lamarckhttp://en.wikipedia.org/wiki/Jean-Baptiste_Lamarckhttp://en.wikipedia.org/wiki/Jean-Baptiste_Lamarck


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2.1.2 MORPHOLOGY OF GAS.

This species is known to have a globular shell in shape. Normal coloration typically

includes bands of brown, black, and yellowish-tan; color patterns are extremely

variable. Albino and gold color variations exist. The size of the shell is up to 150 mm in length

(Howells, R.2008). Apple snails have a dextral shell except the apple snails from the

genusLanisteswho have a sinistral shell (shell opening at the left), although there are reports of

sinistral animals in the other genera (CAZZANIGA, N. J. & ESTEBENET, A. L. 1990)

The surface of the shell can be smooth or rough, depending of the species and the

environment. Malleation (hammered surface) occurs mainly when the snail grows fast. In such

cases the anorganic periostracum, which is first created, is very thin and wrinkles before it

becomes enforced with the rigid ostracum and hipostracum. Stria (growth lines) are formed at

slower growth. New shell material is deposited bit by bit at the shell opening, creating the a new

small ridge or stria with every new piece added. The shell lip (the margin of the shell at the shell

opening) is often thickened in mature snails, while it is sharp in younger snails. The lip is curved

at the outside, while rather straight near the umbilicus. The operculum (the shell door) is a

corneous plate (except in the genusPila, where the operculum is calcified at the inside during the

life of the snail), with a concentric structure and a nucleus near the parietal margin (close to the

umbilicus). With this shell door, the snail is able to close of its shell to survive periods of drought

and as protection against predators (KEAWJAM, R. S.& Upatham, E.S. 1990.).
http://en.wikipedia.org/wiki/Albinohttp://www.applesnail.net/content/lanistes.htmhttp://www.applesnail.net/content/lanistes.htmhttp://www.applesnail.net/content/lanistes.htmhttp://www.applesnail.net/content/anatomy/shell.php#structurehttp://www.applesnail.net/content/anatomy/shell.php#structurehttp://www.applesnail.net/content/pila.htmhttp://www.applesnail.net/content/pila.htmhttp://www.applesnail.net/content/pila.htmhttp://www.applesnail.net/content/pila.htmhttp://www.applesnail.net/content/anatomy/shell.php#structurehttp://www.applesnail.net/content/anatomy/shell.php#structurehttp://www.applesnail.net/content/lanistes.htmhttp://en.wikipedia.org/wiki/Albino


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2.1.3 DISTRIBUTION OF GAS.

Apple snails inhabit a wide range of ecosystems from swamps, ditches and ponds to lakes

and rivers. Not every species has similar preferences. However, most apple snails prefer lentic

waters above turbulent water (rivers) (PERERA, GLORIA & WALLS, JERRY 1996). The

golden apple snail (Pomacea canaliculata Lamarck) is a freshwater prosobranch indigenous to

South America. In the 1980s it was brought in from Argentina to Taiwan for commercial

production as a food source (Mochida, 1991). From Taiwan the snail was distributed to the third

world countries for backyard rearings to generate side-incomes (Anderson, 1993). The estimated

snail-infested areas were 171,425 ha in Taiwan in 1986, 16,195 ha in Japan in 1989 and 400,000

ha in the Philippines in 1989 (Mochida, 1991).


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2.1.4 POTENTIAL OF GAS.

The GAS flesh has a high protein content, which means it can be useful as a locally

available feed resource for monogastric livestock (Lampheuy Kaensombath and Brian Ogle

2006). Ensiled and fresh Golden Apple Snail (GAS) flesh as replacement for fish meal had

nonegative effects for fattening pigs in terms of daily weight gains and feed conversion ratios.

However, feed dry matter intakes were lower when GAS meal replaced fish meal. The cost of the

diet with ensiled GAS flesh was lower than of the diets with fish meal and fresh GAS, including

the labor cost of processing the snails. The snails in fresh or ensiled form in diets for growing

pigs can be profitable if the farmers collect and process the snails themselves, and there should

be an additional benefit of higher rice yields (Lampheuy Kaensombath and Brian Ogle 2006).

Apple snails are well edible and are often considered a protein rich delicacy. Consuming these

snails is therefore an interesting option in those areas where they have become a pest and treat

for the rice and taro production. In such cases, eating these snails has two benefits that is

collecting the snails is encouraged and the diet of the farmers (especially in thirth world

countries) is enriched with a protein source. (Stijn A. I. Ghesquiere 1998). GAS appear in large

numbers and are a potential feedstuff for on farm feed preparation but not for industrial

manufacturing of aquaculture feed. This snail used as fish feed traditionally in Korea, Indonesia,

Thailand (Ravindra C. Joshi). Feeding trial on Nile Tilapia in aquaria showed that GAS meat

meal at 75-100% of the diet mixed with rice bran was beneficial (Cagauan and Doria, 1989)

GAS has a high nutritive value and could be used to completely replace fish meal in African

catfish production (O Phonekhampheng 2008).
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2.2 INTRODUCTION OF Pangasius sutchi

2.2.1 TAXONOMY OF Pangasius sutchi

Kingdom : Animalia

Phylum : Chordata

Class : Actinopterygii

Order : Siluriformes

Family : Pangasiidae

Genus : Pangasius

Species: P. hypophthalmus

Binomial name Pangasius hypophthalmus(Sauvage, 1878)

Synonyms Pangasius sutchi Fowler, 1937
http://en.wikipedia.org/wiki/Animalhttp://en.wikipedia.org/wiki/Chordatehttp://en.wikipedia.org/wiki/Actinopterygiihttp://en.wikipedia.org/wiki/Siluriformeshttp://en.wikipedia.org/wiki/Pangasiidaehttp://en.wikipedia.org/wiki/Pangasiushttp://en.wikipedia.org/wiki/Binomial_nomenclaturehttp://en.wikipedia.org/wiki/Binomial_nomenclaturehttp://en.wikipedia.org/wiki/Sauvagehttp://en.wikipedia.org/wiki/Synonym_(taxonomy)http://en.wikipedia.org/wiki/Synonym_(taxonomy)http://en.wikipedia.org/wiki/Sauvagehttp://en.wikipedia.org/wiki/Binomial_nomenclaturehttp://en.wikipedia.org/wiki/Pangasiushttp://en.wikipedia.org/wiki/Pangasiidaehttp://en.wikipedia.org/wiki/Siluriformeshttp://en.wikipedia.org/wiki/Actinopterygiihttp://en.wikipedia.org/wiki/Chordatehttp://en.wikipedia.org/wiki/Animal


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2.2.2 Pangasius sutchi MORPHOLOGY.

The sutchi catfish is a popular fish for aquaculture; it is cultured widely in Asia in

countries such as Vietnam, Malaysia, Indonesia, Laos, Cambodia, and China. This fish have

long body, latterly flattened with no scales. Head relatively small. Mouth broad with small sharp

teeth on jaw, vornerine and palatal bones. Eyes relatively large. Two pairs of barbels, upper

shorter than the lower. Fins dark grey or black. Six branched dorsal-fin rays. Gill rakers normally

developed. Young fish have black stripe along lateral line and another long black stripe below

lateral line; large adults uniformly grey but sometimes with greenish tint and sides silvery. Dark

stripe on middle of anal fin; dark stripe in each caudal lobe; small gill rakers regularly

interspersed with larger ones (Gustiano, R. 2003).

2.2.3 DIET OF Pangasius sutchi

P. sutchi is omnivorous, feeding on algae, higher plants, zooplankton, and insects, while

larger specimens also take fruit, crustaceans and fish (Griffiths et al.2010). The farm made diet

for Pangasius sutchi is primarily compose of several kind of locally available by-product.

Farmer also use formulated pellet when local by-product was not available(C D Webster 2002).

Ingredient %

Fish meal 15

Soybean meal 15

Groundnut 24

Ricebran 30

Broken rice 15Vitamin and mineral premix 1

Table 1: Model diet formulae for Pangasius (C D Webster 2002).


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2.2.4 POTENTIAL OF Pangasius sutchi

P. hypophthalmus, are important targets of commercial fisheries, such activities are

considered to have led to the decline of stocks in recent years (Allan et al. 2005). P.

hypophthalmus is presently cultured in several countries, including Thailand (Na-Nakorn et

al.2006) and Vietnam (Phuong et al.2008). iet Nam exports P. hypophthalmus to over 80

countries, including several in Europe (especially Poland and Spain), Asian countries, Mexico,

Australia, the United States of America, and the Middle East. New markets such as Russia are

emerging. The European Union remains the most significant market (35 percent by volume, 40

percent by value).Viet Nam has the capacity to process 3 500 tonnes of aquatic product daily and

the capacity is increasing. There are presently 405 industrial-scale processing plants in Viet

Nam, of which 301 are certified for export to Europe and 30 are certified to export to the Russian

Federation;16 percent are currently ISO certified (Griffiths,2010).

FIGURE 1: Global aquaculture production of Pangasius hypophthalmus

(FAO Fishery Statistic)
http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR1http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR1http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR1http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR21http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR21http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR21http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR26http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR26http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR26http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR26http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR21http://www.springerlink.com/content/2451u75tp25lq011/fulltext.html#CR1


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CHAPTER 3

MATERIAL AND METHODS

3.1 STUDY LOCATION

The study was conducted at the freshwater hatcheries of the Universiti Malaysia

Terengganu and labrarotary (UMT) between July 2010 and February 2011.

3.2 GAS MEAL PREPARATION

GAS were collected from FELCRA oil palm plantation from Kg. Kuala Tahan , Jerantut ,

Pahang, Malaysia. There are many methods to process GAS meal first is: Snail was put in clean

water without feeding for 48 hour to make sure intestine is emptied. GAS boiled in water for 15-

20 minutes. The flesh was separate from it shell minced and dried at temperature not more than

0 (Basa S.S 1988). A second method is the shell was broken and removed, and the remaining

cover and flesh cleaned with water before chopping and grinding (O Phonekhampheng et al

2008). The third method is the shells and cover of the snails were removed and then the flesh

washed with clean water and drained. The flesh was then chopped into small pieces of 0.51.0

cm in size. Then both were weighed and mixed in the proportion of 10% of molasses to 90% of

rice bran, on a fresh basis (Lampheuy Kaensombath et al 2004).


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3.3 PROXIMATE ANALYSIS

The proximate chemical composition of feed ingredients were estimated by the methods

described by the (AOAC, 1984), to determine the crude protein content. Proximate analysis

would done to 6 different sample such as GAS meal, pellet GAS0, GAS25, GAS50, GAS100,

GAS100B. This phase would focuses in determination of moisture and dry matter, ash, crude

lipid, crude protein, crude fiber.

3.4 PELLET PRODUCTION.

Five type of experimental fish pellet (GAS0 (control), GAS25, GAS50 ,GAS100

,GAS100B ) were formulated (Table 2). The diets were prepared by mixing all dry ingredients in

a mixer and subsequently adding the wet ingredients to form moist dough. A dough steaming for

15 minutes and will put to forming machine to form a favorable shape to feed. Feed were dry

inside oven under temperature 60C for 12-24 hour. Dried feed were cut using hand to make it

smaller.

Ingredient/ratio(fish meal :GAS meal

0%(control) 25% 50% 100% 100+shell%

Fish meal 140 g 105 g 70 g 0 g 0 g

GAS meal 0 g 35 g 70 g 140 g 140 g

Soy bean meal 466.9 g 466.9 g 466.9 g 466.9 g 466.9 g

Rice bran 37.1 g 37.1 g 37.1 g 37.1 g 37.1 g

Wheat flour 21 g 21 g 21 g 21 g 21 gFish oil 21 g 21 g 21 g 21 g 21 g

Mix vitamin 7 g 7 g 7 g 7 g 7 g

Mineral 7 g 7 g 7 g 7 g 7 gTable 2: pellet ingredient


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3.5 FISH MANAGEMENT

3.5.1 TANK DESIGN

Figure 1: Culture system ofPangasius sutchi

3.5.2 CULTURE SYSTEM

Pangasius sutchi fingerlings were obtained from a freshwater hatchery UMT.

Fish from same brood stock will culture in 5 different tanks. Each tank contains 20

Pangasius sutchi fingerlings. Each tank will supply with aeration. Water quality

parameters (temperature, salinity, pH, and dissolved oxygen) during the experimental

period will be observed.

Tank: A (Control)

Fish: 20

Feeding : pellet GAS0%

Tank: B

Fish: 20


Tank: C

Fish: 20


Tank: D

Fish: 20

Feeding : pellet

GAS100%

Tank: D2

Fish: 20

Feeding:

pelletGAS100%s

Aeration : 24 hour

Tank size : 150l


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3.6 FEEDING AND GROW RATES DETERMINATION.Fish in each tank will be give different formulation of pellet GAS0, GAS25,

GAS50, GAS100, GAS100B twice per day in each different tank. Fish will be measure

every week until 1 month duration of experiment.

The following calculations were made on collected data to describe and evaluate fishperformance:

(i) Mean weight gain (MWG) = Wf -Wi

Where, Wf is the mean final fish weight and Wi the mean

initial fish weight.

(ii) Percentage body weight gain (PWG) = (MWG x 100)/Wi

(iii) Specific growth rate (SGR, %) (Brown 1957) = [(log Wflog Wi) x 100)]/D

(iv) Feed conversion ratio (FCR) = feed consumed (dry weight)/fish weight gain

3.7DATA ANALYSISThe data were subjected to analysis of variance (ANOVA) by using the General Linear

Model (GLM) procedure of Minitab version 14 (2000). When the F test was significant

(p


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CHAPTER 4

PROJECT SCHEDULE

Project activities/ week M J J A S O N D J F M

1. Preparation of research proposal. x x2. Sent a research proposal 03.

Preparation of golden apple snail meal x4. Lab set up. 0a) Lab design set up x x

b) Hatcheries design set up. x x

5. Breeding and care of fish for test. x x x6. Proximate analysis for golden apple snail

meal.x

7. Formulation of fish pellet. x x8. Production of fish pellet x x9. Proximate analysis of each pellet x10.Determination of growth rates of fish given

different pellet.

0

a) Run experiment and data collection x x11.Determination of water quality in different

tank.x x

a) Run experiment and data collection 012.Data analysis x x13.Report writing x x14.Send report. 0

x : Activities 0: Planned milestone

Year 2010 2011


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CHAPTER 5

MILESTONE

Date Progress

3/8/2010 Completing first draft proposal

Setting up experiment

1st week of August Preparing GAS meal.

2nd week of August Preparing formulated diet

3rd week of August Introduce fish to tank.

Feeding experiment

Water quality sampling

4th week of August

1st ,2nd week of September

Proximate analysis of GAS meal and each

formulated feed.

22th of August 1st sampling

29th of August 2nd sampling

5th September 3rd sampling

12th September 4th sampling

13th September Cleaning tank and experimental stuff.

4th week of September

1st and 2nd week of October

Data analysis

3rd week of October onwards Completing thesis.


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CHAPTER 6

REFERENCES

Allan JD, Abell R, Hogan Z, Revenga C, Taylor BW, Welcomme RL, Winemiller K (

2005) Overfishing of inland waters. Bioscience 55:10411051

ANDERSON, B. 1993. The Philippines snail disaster. The Ecologist23, 7072.

Australian Centre for International Agricultural Research ACIAR

Basa S.S 1988 : COUNTRY REVIEW

Caugauan,A.G, and L.S.Doria1989 golden apple snail meal as feed for Nile tilapia

fingerling in aquaria.CLSU Scientific journal.9(3)

CAZZANIGA, N. J. & ESTEBENET, A. L. 1990. A

sinistral Pomaceacanaliculata (Gastropoda: Ampullariidae). Malacological Review, 13(1/2): 123-143.

C D Webster, Aquaculture Research Center, Kentucky State University, USA, C E Lim,

USDA-ARS, Fish Diseases and Parasites Research Laboratory, Auburn, Alabama,

USA.

Gustiano, R. 2003. Taxonomy and phylogeny of Pangasiidae catfishes from Asia

(Ostariophysi, Siluriformes). PhD thesis. Katholieke Universiteit Leuven, Leuven,

Belgium. 295 pp.

Griffiths, D., Van Khanh, P., Trong, T.Q. FAO. 2010. - . Cultured Aquatic Species

Information Programme. In: FAO Fisheries and Aquaculture

Department[online]. Rome. Updated 16 February 2010. [Cited 3 August 2010].


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Howells, R. Personal communication. Texas Parks and Wildlife Department. In: United

States Geological Survey. 2008. Pomacea canaliculata.USGSNonindigenous

Aquatic Species Database, Gainesville, FL. Revision Date: 2/4/2008

KEAWJAM, R. S.& Upatham, E.S. 1990. Shell morphology, reproductive anatomy and

genetic patterns of three species of apple snail of the genus Pomacea in thailand.

Med. & Appl. malacol., 2: 45-57.

Lampheuy Kaensombath* and Brian Ogle**Laboratory-scale ensiling of Golden Apple

Snails (GAS)(Pomacea spp)Lampheuy Kaensombath* and Brian Ogle Lampheuy

Kaensombath* and Brian Ogle**Faculty of Agriculture, National University of

Laos, Vientiane, Lao PDR Department of Animal Nutrition and Management,

Swedish University of Agricultural Sciences, Box 7024, 75007 Uppsala,

Sweden

Peter Edwards, Le Anh Tuan, Geoff L. Allan A survey of marine trash fish and fish mealas aquaculture feed ingredients in Vietnam ACIAR Working Paper No. 57(printed version published in 2004)

M. L. WINDSOR 2001 DEPARTMENT OF TRADE AND INDUSTRY TORRYRESEARCH STATION Fish Meal.

MOCHIDA, O. 1991 Spread of Freshwater Pomacea snails Pilidae Mollusca from

Argentina to Asia. MICRONESICA 3: 51-62

Na-Nakorn U, Kamonrat W, Ngamsiri T (2004) Genetic diversity of walking

catfish, Clarias macrocephalus, in Thailand and evidence of genetic introgressionfrom introduced farmed C. gariepinus. Aquaculture 240:145163

OPINION 1913 Pila Rding, 1798 and Pomacea Perry, 1810 (Mollusca, Gastropoda):

placed on the Official List, and AMPULLARIIDAE Gray, 1824: confirmed as the

nomenclaturally valid synonym of PILIDAE Preston, 1915

O Phonekhampheng, L T Hung* and J E Lindberg( 2009). Ensiling of Golden Apple

snails (Pomacea canaliculata) and growth performance of African catfish

(Clarias gariepinus) fingerlings fed diets with raw and ensiled Golden Apple

snails as protein source.

PERERA, GLORIA & WALLS, JERRY 1996. Apple Snails in the Aquarium. 121

pages, T.F.H. Publishing Ltd

Phuong NT, Sinh LX, Thinh NQ, Chau HH, Anh CT, Hau NM (2008) Economics of

aquaculture feeding practices: Viet Nam. In: Hasan MR (ed) Economics of
http://en.wikipedia.org/wiki/United_States_Geological_Surveyhttp://en.wikipedia.org/wiki/United_States_Geological_Surveyhttp://en.wikipedia.org/wiki/United_States_Geological_Surveyhttp://en.wikipedia.org/wiki/United_States_Geological_Survey


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aquaculture feeding practices in selected Asian countries, FAO Fisheries

Technical Paper 505. FAO, Rome, pp 183205

Ravindra C. Joshi Philippine Rice Research Institute (PhilRice), Maligaya, Science City

of Muoz, Nueva Ecija 3119, Philippines E-mail:[email protected]

Robert H. Cowie & & IUCN/SSC Invasive Species Specialist Group (ISSG). (Last

Modified: Wednesday, 13 April 2005) Global Invasive Species Database,

2008. Pomacea canaliculata

.

Sison J.a (1985): Hand book on crisis management on feedmeal and technology for the

Philippines. .Feedindex (Phills.),Quezon City. The Philippines.

Stijn A. I. Ghesquiere 1998 http://www.applesnail.net/

Teo, Su Sin1 Golden Apple Snail (Pomacea canaliculata Lamarck, 1819) in Sabah,

MalaysiaCurrent Situation and Management Strategy
mailto:[email protected]:[email protected]:[email protected]://www.applesnail.net/http://www.applesnail.net/http://www.applesnail.net/mailto:[email protected]


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Proximate analysis: Ash

Procedure

1. Accurately weight ca 5 g of sample in a crucible wich has been ignited and tared (use 2.5g of sample in the case of products which have a tendency to swell).

2. Place crucible in drying oven at 100 oC for 24 hours.3. Transfer to cool muffle furnace and increase the temperature step wise to 550 oC 5 oC.4. Maintain temperature for 8 hours or until a white ash is obtained.5. If white ash is not obtained after 8 hours, moisten ash with distilled water, slowly dry on

a hot plate, and re-ash at 550 oC to constant weight. Repeat if necessary.

6. Remove crucible to a desiccator and weight soon after cool.Calculation

Calculate the percentage ash content (wet weight basis) as follows:

(wt. crucible and ash - wt. crucible)

% ASH (wet)= x 100

(wt. crucible and sample - wt. crucible)

Calculation of ash content on dry basis (when moisture content is known) as follows:

% ash (wet)

% ASH (dry)= x 100

(100 - % moisture)


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Proximate analysis: Crude protein

Procedure

1. Accurately weigh a suitable quantity of fine-grained material (ca 1.2 g for fishmeal, ca2.5 g for solubles or homogenized fish) and place in digestion flask.

2. Add sequentially 15 g Na2SO4, 1 g CuSO4, one or two selenized boiling granules and 25mL of conc H2SO4 to the flask.

3. Digest until solution is almost colourless or light green (2 hrs for inorganic material) andthen at least a further 30 minutes. Do not heat any part of the Kjeldahl flask above the

level of the digestion mixture.

4. Cool (do not allow to solidify), and cautiously add 200 mL water. Add additional boilinggranules (if necessary) to prevent bumping.

5. Pipette 100 mL 0.1 N HCl into a 500 mL erlenmeyer flask, add 1 mL Conway's indicatorand place the flask under the condenser ensuring that the condenser tip is immersed in the

acid solution. (volume of standardized HCl used in distillation may be varied according

to the expected nitrogen content of the sample).

6. Tilt the Kjeldahl flask containing the digested sample and add 100 mL of 50% NaOHsolution slowly down the side of the Kjeldahl flask so that it forms a layer underneath the

digestion mixture.Immediately connect the flask to the distilling bulb of the distillation

apparatus. Rotate flask to thoroughly mix contents.

7. Heat until all ammonia has passed over into the standard acid. Collect approximately 150mL. Caution, flask will bump. Remove immediately (prolonged boiling and too rapid

distillation of acid during digestion should be avoided as loss of ammonia may occur).

8. Wash tip of condenser and titrate excess standard HCl in distillate with NaOH standardsolution (detailed titration procedure) .
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Calculation

Calculate the percentage nitrogen (wet weight basis) as follows:

(A - B) x 1.4007

% Nitrogen (wet) = x 100

weight (g) of sample

where:

A = vol. (mL) std. HCl x normality of std. HCl B = vol. (mL) std. NaOH x normality of std. NaOH

Calculate nitrogen content on dry basis (when moisture content is known) as follows:

% Nitrogen (wet)

% Nitrogen (dry)= x 100

(100 - % moisture)

Calculate the percentage protein (wet or dry basis) as follows:

% PROTEIN = % nitrogen x 6.25

where 6.25 is the protein-nitrogen conversion factor for fish and fish by-products.


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Proximate analysis: Crude lipid

Extraction:

1.

Line the fat beakers up in front of the extractor and match the thimbles with theircorresponding fat beakers.

2. Slip the thimble into a thimble holder and clip the holder into position on the extractor.3. Add about 40 mL of diethyl ether (one glass reclaiming tube full) to each fat beaker.4. Wearing white gloves, slip the beaker into the ring clamp and tightly clamp the beaker

onto the extractor. If the clamp is too loose, insert another gasket inside the ring.

5. Raise the heaters into position. Leave about a 1/4 inch gap between the beaker and theheating element.

6. Turn on the heater switch, the main power switch, and the condenser water.7. After the ether has begun to boil, check for ether leakage. This can be detected by

sniffing around the ring clamp. If there is leakage, check the tightness of the clamp and if

necessary replace the gasket(s).

8. Extract for minimum of 4 hr on a Hi setting (condensation rate of 5 to 6 drops persecond), or for 16 hr on a Low setting (condensation rate of 2 to 3 drops per sec).

9. After extraction, lower the heaters, shut off the power and water, and allow the ether todrain out of the thimbles (about 30 min). This is a good stopping point.

Destillation and weighing

1. Remove the thimble from the holder, and rinse the holder with a small portion of diethylether from the washbottle. Clip an ether reclaiming tube in place and reattach the fat

beaker.

2. Reposition the heaters and turn on the electricity and water. Proceed to distill the etherusing a Hi setting. Watch Closely.

3. Distill until a thin layer of ether remains in the bottom of the beaker, and then lower theheater. Do not allow beakers to boil dry. Overheating will oxidize the fat. When the last

beaker has finished, shut off the power and water.


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4. Wipe the exterior of the beaker clean with a Kimwipe as it is being removed from theextractor.

5. Empty the reclaiming tubes into the "USED" diethyl ether container.6. Place the tray of beakers in an operating hood to finish evaporating the ether. If there is

no hurry, air moving through the hood will be sufficient without heat. A steambath may

be used to speed up the evaporation. Beakers should remain in the hood until all traces of

ether are gone. Carefully sniff each beaker to determine if any ether remains.

7. Place the beakers in a 102C gravity convection oven. Warning: If a beaker containingether is placed in the oven an explosion may occur.

8. Dry for 1/2 hr. No longer. Excessive drying may oxidize the fat and give high results.9. Cool in a desiccator and weigh and record weight to the nearest 0.1 mg (W2). The fat

beakers are best cleaned by warming on a steambath or on a hot plate on a low setting.

Add some used ether to dissolve the fat. The use of a rubber policeman is helpful.

10.After soaking the beakers in Alconox detergent, wash them using hot water and vigorousbrushing. The thimbles are best cleaned by blowing out with air.

If doing a proximate analysis, the residue left in the thimble may be used to determine crude

fiber.

Calculation

Percent Crude Fat (Ether Extract), DM basis:

(AWres - Wta)

% Crude fat (wet) = x DM (%)

weight (g) of sample

Wta = tare weight of beaker in grams Wres = weight of beaker and fat residue in gram


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Proximate analysis: Crude fiber

Procedure

1. Determine separately the sample moisture by heating in an oven at 105 C to constantweight. Cool in a desiccator.

2. Weight accurately 1 g about of grinded sample (1 mm about) approximately with 1 mg.==> W1

3. Add 1.25% sulfuric acid up to the 150 ml notch, after preheating by the hot plate in orderto reduce the time required for boiling.

4. Add 3-5 drops of n-octanol as antifoam agent.5. Boil 30 minutes exactly from the onset of boiling.6. Connect to vacuum for draining sulfuric acid.7. Wash three times with 30 ml (crucible filled up to the top) of hot deionized water,

connecting each time to compressed air for stirring the content of crucible.

8. After draining the last wash, add 150 ml of preheated potassium hydroxide (KOH) 1.25%and 3-5 drops of antifoam.

9. Boil 30 minutes.10.Filter and wash as point 7.11.Perform a last washing with cold deionized water aimed to cool the crucibles and then

wash three times the crucible content with 25 ml of acetone, stirring each time by

compressed air.

12.Remove the crucibles and determine the dry weight after drying in an oven at 105 C foran hour or up to constant weight. Let cool in a desiccator. This weight (W2)represents the

crude fiber plus ash content in comparison to initial weight.

Calculation

Calculate the percentage crude fiber (wet weight basis)as follows:


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(W2 - W1)

% Crude fiber (wet) = x 100

W1

Proximate analysis: Nitrogen Free Extract (NFE)

% NFE = % DM - (% EE + % CP + % ash + % CF)

where: NFE = nitrogen free extractDM = dry matter

EE = ether extract or crude lipid

CP = crude protein

CF = crude fiber

2)

GROWTH PERFOMANCESThe following calculations were made on collected data to describe and evaluate fishperformance:(i) Mean weight gain (MWG) = Wf -Wi

Where, Wf is the mean final fish weight and Wi the meaninitial fish weight.

(ii) Percentage body weight gain (PWG) = (MWG x 100)/Wi

(iii) Specific growth rate (SGR, %) (Brown 1957) = [(log Wflog Wi) x 100)]/D

(iv) Feed conversion ratio (FCR) = feed consumed (dry weight)/fish weight gain


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