Alternative Fish Feeds

Project IV: Development and evaluation of alternative feed source for
aquaculture
Background and justification
Several attempts have been made to find alternate fish feeds from various sources throughout the
world. These feed sources could originate from either plant or animal sources. Under this project
different feed sources such as the aquatic fern Azolla spp., the vermicomposting earthworm,
Eisenia spp. the cladoceran Brachionus and Daphnia spp., as well as the blue green currently
known as Arthrospira fusiformis ( previously Spirulina platensis) will be evaluated as direct feed
or feed ingredients for fish.
Azolla (association of fern and cyanobacteria) is found to be the most economic and sustainable
animal feed substitute in India, China, Japan, Latin and South America (Chatterjee et al., 2013).
It has high source of protein and thereby a potential feed ingredient for animals (Kamalasanana
et al., 2002). Azolla is a small aquatic floating fern that lives in symbiosis with the nitrogen
fixing blue-green algae ( Anabaena azollae) which has a high nitrogen fixing ability (Van-Hove,
1989).
Vermicomposting is using worms to recycle food scraps and organic matter into a valuable soil
amendment. However, this study aims at propagating the worm in mass for fish feed. Culturing
of the worm requires the need of worms such as bin, container, organic matter (cow/donkey
dung) as feed. The worm needs moisture, air, food, darkness and warm temperature (10-25oC.)
Bedding is important made of newspaper, leaves or for holding moisture and space. The bedding
place can be on the ground or made from wood or concrete. The two worms which are widely
used in vermicultre include Eisenia fetida and Eisenia andredi. These two earthworm species
have been imported and cultured at Ambo Plant Protection Center. This technology has been
imported from Russia through the JERBE group which is one of the collaborative research
project run in collaboration with Russian scientists.
The blue green alga, Arthrospira fusiformis has been widely used for different purposes
including as animal feed and fodder. In nature, Arthrospira fusiformis have been reported from
highly saline and alkaline lakes such as Lakes Abijata, Arenguade and Chitu (Elizabeth Kebede,
1996). In these lakes Spirulina is produced as unialgal mat constituting over 90% of the algal
production and productivity. This alga has been reported to be rich in protein constituting about
60% crude protein (references).
One of the major bottle necks in culturing of fish is the availability alternative live feeds for
larvae and juvenile fish. Currently the brine shrimp Arthemia spp. have been used widely all over
the world. However, despite its importance the seed form is imported from abroad and costs hard
currency which is not affordable by farmers. Therefore, there is a strong need to culture and mass
produce alternate sources from zooplanktons. The rotifer Brachionus spp and the cladocerans are
considered as alternate substitutes to tackle the problem.
Objectves/s
General objective
 To develop fish feeds originating from plant and animal sources for adult and juvenile
fish which can be used as substitutes for enhancing fish culture.
Specific objectives
 To adapt Azolla harvesting techniques and produce a cheap as well as a high quality fish
feed or tilapia.
 To adapt mass propagation nutrient, contents of earthworms and test its use as fish feed
for juvenile catfish.
 To adapt mass culturing technique for rotifers and cladoceran to be used as live feed for
fish larvae
There are seven activities that are included under this project which is listed below.
1. Use of a natural aquatic fern (Azolla spp.) − as a main component of feed for Tilapia
2. Use of vermiculture as direct live feed and ingredient of fish feed for catfish.
3. Evaluate culturing techniques of the freshwater rotifer, Brachionus calyciflorus and its
application in fish larviculture.
4. Effect of fish silage addition into the diets on the growth performance of the African
catfish.
5. Comparative assessment of proximate composition & mineral content in farmed and wild
tilapia.
6. Isolation and culturing of Arthrospira fusiformis (Spirulina platensis) in laboratory scale.
7. Comparisons of periphyton and feed-based ponds for water quality and growth
performance of the Nile tilapia, Oreochromis niloticus in ponds
Activity 1: Use of a natural aquatic fern (Azolla sp.) as a main component of feed for
Tilapia
Introduction
Several attempts have been made to find alternate sources of animal feed throughout the world.
Azolla (association of fern and cyanobacteria) is found to be the most economic and sustainable
animal feed substitute in India, China, Japan, Latin and South America (Chatterjee et al., 2013).
It has high source of protein and thereby a potential feed ingredient for animals (Kamalasanana
et al., 2002). Azolla is a small aquatic floating fern that lives in symbiosis with the nitrogen
fixing blue-green algae ( Anabaena azollae) which has a high nitrogen fixing ability (Van-Hove,
1989). Out of several species of Azolla, Azolla microphylla and Azolla pinatea have been
reported to be best suited for tropical climate feed ingredient for cattle, poultry, fish, sheep, goats
and pigs (Chatterjee et al., 2013, Fiogbe et al., 2003). Currently, it is widely used as an income
generating cheep aquatic plant for commercial and local farms in India (das et al., 2005,vk-
nardep, 2010 ). It grows naturally in stagnant water of drains, canals, ponds, rivers, marshy lands,
and lake basins and it is is an easily propagated aquatic plant in a standing water with a relative
humidity of 85-90%, pH of 4.5-6.5, salinity of between 90-150 mg/L and adequate phosphorus
for its nutritional needs.
Azolla doubles its weight in 3-5 days, from a start of 1t/ha, it can reach a fresh weight of 15-20
t/ha in about 20 days (Basak et al., 2002). The plant is very rich in protein and minerals, on a dry
weight basis; it contains 25 - 35 percent protein, 10 - 15 percent minerals and 7 - 10 percent of
amino acids, bio-active substances and bio-polymers (Van- Hove, 1989). Its high nutrient
composition makes it a highly efficient and effective feed for livestock, poultry, fish and pigs
because of it is easily digestible, owing to high protein and low lignin content. Azolla can be
mixed with feed concentrates or can be given directly to livestock, poultry, sheep, goats, pigs and
rabbits (Vk-Nnardep, 2010). A research conducted by Murthy et al., 2013 showed that 2 Kg
fresh Azolla could replace 50% of concentrate cow feed and resulted in 18.5 % higher milk
production while the feed cost was cut by 16.5 %.. Another research conducted Rai et al., 2012
also showed that Azolla feed given to chickens had gained higher body weight and egg
production.
Livestock, poultry and fish farms in Ethiopia are the most lucrative business which have become
a subsistence economic and food sources for about 85% of the population in the country. The
government has given a special attention for the sector considering that Ethiopia has a huge
economic potential of livestock, poultry and fish resources if the people practices a modern and
an organized way of farming. The government is currently organizing small enterprises and
households in one of the three business areas to change their life thereby to bring an overall
economic growth in the country. Recently, the business sector has become a rapidly developing
enterprise area so that large numbers of farms are being established in different parts of the
country, which create significant employment opportunities to many people and bring foreign
currency. However, the sector couldn't move as needed because of feed scarcity and high cost.
The feed cost incurred about 60-65% of the total cost of culture fish. Availability of quality feed
at a reasonable cost is a key to successful operation. The conventional and open field sources of
animal feeds couldn't be enough to mitigate the shortage of feeds and fodder and to make animal
production viable and profitable. It would therefore be wise to use other cheep alternative feed
to the diet formulation to reduce the production cost for livestock, poultry and fish. This project
idea is adapted mainly from India which never been tried so far in Ethiopia.
Objective/s
General objective
 To adapt Azolla harvesting techniques and produce a cheap as well as a high quality fish
feed or tilapia.
Specific objectives:
 To collect and culture Azolla species in laboratory and open field.

 To adapt mass production technique of Azolla spp. Under controlled condition
 To mass produce Azolla in large amoung to be used as fish feed ingredient for tilapia.
Materials and Methods
Phase one: Azolla sample collection, identification and harvesting activities
Azolla mother seeds from different aquatic environments such as Lakes Ziway, Hawassa and
swampy areas in Deberezeit will be collected in 20 liters plastic jars with 1g of Super Phosphate
or some amount of cow- dung. Azolla mother seeds will be grown in plastic backet/ tray at room
temperature with cow dungs nd other chemicals according to Stanier et al., 1981. Types of the
Nutrient contents of each azolla species will be determined at the Sebeta National Fisheriies &
other Aquatic Life Research Center.
Phase two: Proximate chemical composition and fish feeding trial using Azolla as
ingredient
Following identification and nutrient analysis, optimal growth factors for Azolla microphylla
and/or Azolla pinatea will be determined. Other Azolla species may also be used based on their
nutrient analysis. A number of Azolla microphylla and Azolla pinatea in ponds or plastic trays.
Optimal growth conditions of the Azolla microphylla and Azolla pinatea will be determined
using cheep phosphate nutrient rich sources like cow-dung and poultry wastes considering the
applicability of the project at local farms and household level. Biomass of the Azolla species will
then be harvested and prepared for fish feed in fresh and pellet form. The feed trial will be
conducted either in ponds or tanks at Sebeta National Fisheries and Aquatic Life Research
Center. The Azolla feed will be given to the experimental fish containing different proportions of
the dried Azolla feed. The effect of the various feeds will be evaluated every two weeks. The
growth rate of fish and other water quality parameters will be recorded regularly. Parameters
such as fish total weight, total length of fish will be recorded from each treatment including the
control.
Location
National Fisheries & Aquatic Life Research Center (NFALRC), Sebeta
Duration
2008 . – 2010 E.C.
Work plan
Activity Months
Jul Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun
Use of a natural aquatic fern as

a main component in feed for
tilapia
Dig small ponds for fern x x

propagation
Transfer of collected ferns into x x x x x x x x x x

small ponds
Mass propagation and care of x x x x x x x x

ferns in ponds
Purchase of plastic trays for fern 01-15 01-15

culture
Drying & Chemical analysis of x x x

ferns
Feed formulation for feed x x

experiment
Activity Months
Collection of male tilapia x

fingerlings
Feeding of tilapia with fern feeds x x x x x x
Progress report 23 23 23 23
Budget by code for 2009 E.C.

Code Birr
6114 12179.31
6212 6336.945
6213 3168.473
6215 5280.788
6217 11089.65
6218 8660.492
6219 8660.492
6223 4752.709
6231 61257.14
6232 5280.788
6241 12673.89
6243 5280.788
6244 8713.3
6245 14786.21
6256 4752.709
6313 16898.52
Total 189772.2
Expected output
 A mass production technology of Azolla spp. Will be adopted under controlled system.
 A cheap and high quality animal feed in the form of dried and/or fresh Azolla will be
produced to be used as fish feed for tilapia.
Responsibilities
Responsible persons: Zenebe Tadesse, Adamneh Dagne, Abelneh Yimer & Bizuayehu
National Fisheries & Aquatic Life Research Center
Monitoring and evaluation matrix
Method of Tools for

M & E Information to be data collecting the Method of
Out put Objectives Indicators collected collection information analysis
-
Activities
-
Inputs
-
REFERENCES
 Basak B, Pramanik A H, Rahman M S, Tarafdar S U and Roy B C. ( 2002) . Azolla
(Azolla pinnata) as a Feed Ingredient in Broiler Ration. International Journal of Poultry
Science 1 (1): 29-34,
 Chatterjee A, Sharma P, Ghosh M K, Mandal M and Roy P K. (2013). Utilization of
Azolla Microphyllaas Feed Supplement for Crossbred Cattle. International Journal of
Agriculture and Food Science Technology.ISSN 2249-3050, Volume 4, Number 3
(2013), pp. 207-214
 Das D., Sikdar K., Chetterjee A K.( 2005). Potential of Azolla pinnataas biogas
generator and as a fish-feed. Indian J. Environm. Health, vol. 36, pp. 186-191.
 Ethiopian investment bureau. (2012). A draft proposals for 200 selected priority
investment areas for local and international investors. Addis Ababa, Ethiopia
 Fiogbe E D, Micha J C and Van Hove. (2003). Use of a natural aquatic fern, Azolla
microphylla, as a main component in food for the omnivorous– phytoplanktonophagous
tilapia, Oreochromis niloticus L. J. Appl. Ichthyol. 20 (2004), 517–520
 Kamalasanana Pillai, Premalatha, S. &Rajamony, S. (2002). Azolla – A sustainable feed
substitute for livestock. Leisa India, March, 2002
 Murthy, T N , Ashok M, Thirumalesh T, Umesh B U, Nataraju O R .( 2013). Effect
of partial replacement of AZOLLA for concentrate supplement on lactating
crossbred cows. Environment and Ecology, 31 (2): 415-417
 Rai R B, Dhama K , Damodaran T, Hamid A, Sweta R, Balvir S, Bhatt P..( 2012).
Evaluation of azolla (AZOLLA PINNATA) as a poultry feed and its role in poverty
alleviation among landless people in northern plains of India. Vet. Pract., 13 (2):
250-25.
 Stanier R., Ku-Nisawa R., Mtandel M., Cohen-Bazire G. (1971). Purification and
properties of unicellular blue-green algae (order Chroococcales). Bacteriol.Rev.35:71-
205.
 Van- Hove C. ( 1989). Azolla and its multiple uses with emphasis on Africa. Food and
Agriculture Organization, Rome
 vk-nardep. 2010 Azolla Backyard Cultivation: vknardep.org/services/sustainable-
agriculture/81-azolla-for-the-rescue.html accessed on 11.05.2011
 Watanabe 1. (1998). Azolla and its use in lowland rice culture. Soil and Microbes, Japan,
20, 1-10.
Activity 2: Use of vermiculture as direct live feed and ingredient of fish feed for catfish
Introduction
Vermicomposting is using worms to recycle food scraps and organic matter into a valuable soil
amendment. However, our main target is mass propagation of the worm for fish feed. Culturing
of the worm requires the need of worms such as bin, container, organic matter (cow/donkey
dung) as feed. The worm needs moisture, air, food, darkness and warm temperature (10-25oC.)
Bedding is important made of newspaper, leaves or for holding moisture and space. The bedding
place can be on the ground or made from wood or concrete. The two worms which are widely
used in vermicultre include Eisenia fetida and Eisenia andredi. These two earthworm species
have been imported and cultured at Ambo Plant Protection Center. This technology has been
imported from Russia through the JERBE group which is one of the collaborative research
project run in collaboration with Russian scientists.
The vermicompost which are practiced at APPC aimed at producing natural compost which
serves as natural fertilizers for the local farmers. However, we are using this vermicompost as a
means to mass propagate the worm and use it as live feed or ingredient of fish feed for catfish.
Larvae and juvenile fish are known to feed on live feeds at the early stage of their life cycle.
Fish feed accounts for at least 40 - 60% of the total cost of fish production (Craig and Helfrich
2002; Jamu and Ayinla 2003). In order to enhance aquaculture production, improve food
security, and reduce the level of poverty in developing countries, a search for inexpensive and
locally available feedstuffs is required. However, in Ethiopia a number of by-products from
agricultural processing is available, which are usually not utilized for human consumption, but
may have a high potential for small-scale aquaculture.
Development of a feed for fish production involves evaluation of proximate composition of feed
components, digestibility and performance efficiency as well as cost implications and conditions
of application. Therefore, this activity will be conducted to determine the proximate composition
of selected locally available feed components for small-scale aquaculture feed development. A
specific focus was put on currently underutilized animal protein sources, agricultural and agro-
industrial by-products which are not consumed by the human population. Data from the current
study are also expected to form the basis for further evaluation of the effects of selected feed
components on digestibility and fish growth under different culture conditions.
Objective/s
General objective
To adapt mass propagation nutrient, contents of earthworms and test its use as fish feed for
juvenile catfish.
Specific objectives
 To adapt the culture and mass production techniques of earth worms.
 To determine proximate composition of the worms and compare with other fish feeds in
of animal origin.
 To formulate fish feed using worms as animal ingredients of the feed mixes for feeding
trials.
 To experimentally test the acceptability of live worms and importance of dried

formulated feeds on the growth of juvenile catfishes.
Materials and methods
The following are some of the chemical and experimental trials that will be conducted on station
and on farm at farmer’s yard. Proximate chemical contents of single and mixed feeds will be
determined using standard techniques (AOAC, 2005). Based on the results of chemical
ingredients cheap feeds will be formulated for catfish. Moreover a series of aquaria feeding
experiments will be conducted on stations.,
Experimentt 1. Mass culture and production of worms
The initial inoculum worm (Eisenia vetida) will be transported from Ambo Phyopathology
Research Center to Sebeta, National Fisheries & Aquatic Life Research Center. The starting
inoculums will be further propagated in a series of plastic trays and adapt the propagation
technique.
Culturing of the worm requires the need of worms such as bin, container, organic matter
(cow/donkey dung) as feed. The worm needs moisture, air, food, darkness and warm temperature
(10-25oC.) The bedding place will be either on the ground or made from wood or concrete. For
this study we construct concrete bedding on the ground which serves as inoculums source and a
series a plastic trays for production of worms to be used for fish feeding trial. The efficiency of
cow and donkey dung mixed with wheat bran or brewery wastes will be compared
Experiment II. Proximate composition of dried worms and live feeding trial tom catfish
Proximate composition of dried worms will determined following standard procedures (APHA,
2000). Crude protein will be determined using Kildhal methods after digestion and acid
hydrolysis of a known weight of dried sample. The protein will be estimated by multiply the
level of nitrogen by a factor of 6.25. Total lipid will be determined with standard solvents (ethyl
ether) using soxhlet apparatus. The carbohydrate content will also be determined colorimetrically
(Strickland & Persons, 1986). The acceptance of live worms by juvenile fish will be tasted in the
hatchery using catfish. The feeding will be given at three levels (3%, 6% and 10%) of the fish
total body weight. The feeding experiment will be conducted in triplicate in aquaria indoor.
Experiment III. Test of formulated feeds of worms as substitute for fish meal
Three dried formulated feeds will be prepared containing different proportion of dried earthworm
as indicated below. Similarly, corresponding proportions of three feeds with tilapia carcass will
be prepared for comparison. A total of six feeds will be formulated for the feeding trial.
 10% with worm, with tilapia carcass

The feeding experiment will be conducted in triplicate in aquaria indoor using catfish juveniles.
1. Location
The study will be conducted at Sebeta National Fisheries & Aquatic Life Research Center
2. Duration
Three years, 2008 EC – 2010 EC.
Work plan for 2009

Activity Months
Use of earthworm as feed

ingredient for catfish
Purchase of construction X x x x x
materials for vermiculture
shelter
Construction of shelter house x x x X

for vermiculture
Mass propagation of X x x x x X x x x x
earthworms
Proximate chemical analysis of x x X

earthworms
Collection of catfish fingerling x X

and acclimation to pond/tank
Live feeding of catfish x X x x x x

fingerlings with live earth
worms
Activity Months
Monthly measurements of x x x x x x
morphometric papraameters of
fish and pond dynamics
Progress report X x x x
Budget by code for 2009 E.C.
Code Birr
6114 10034
6212 10034
6213 3169
6215 5809
6217 8977
6218 8396
6219 7868
6223 4753
6231 62313
6232 2640
6241 15314
6243 6337
6244 6337
6245 17427
6256 3697
6313 18483
Total 191587
Expected outputs
The project aims at producing intermediate and final out puts that should be evaluate and
monitored along the implementation process. These out puts are time bound and be assed and
evaluated by different stakeholders.
 Mass culturing (Vermicultre) of worms will be adopted.
 Use of worms as single feed will be experimentally tasted.
 Use of worms as fish feed ingredients will be evaluated.
Responsibilities (persons, centers)

Responsible persons: Zenebe Tadesse, Fasil Degefu, Adamneh Dagne, & Abelneh Yimer
National Fisheries & Aquatic Life Research Center (NFALRC), Sebeta

Monitoring and Evaluation Matrix
Method of Tools for

M & E Information to be data collecting the Method of
Out put Objectives Indicators collected collection information analysis
-
Activities
-
Inputs
-
References
 Aguilar-Manjarrez, J. & Nath, S.S. (1998) A Strategic Reassessment of Fish Farming

Potential in Africa. FAO, Rome.
 AOAC (Association of Official Analytical Chemists) International 2005 Official
Methods of Analysis of AOAC International, 18th Edition. Gaithersburg, Maryland,
USA, AOAC International.
 Craig, S and L.A. Helfrich (2002): Understanding Fish Nutrition, feeds and Feeding,
Cooperative Extension Service, publication 420-256. Virginia State University, USA.
 Edwards, P. (1999) Aquaculture and Poverty: past, present and future prospects of
impact. SIFAR, Rome.
 FAO (2003) Fisheries statistics, http://www.fao.org. Accessed 13th Jan.2006.
 FAO (2010) State of world fisheries and aquaculture 2020, Fisheries Technical Paper
ISSN 1020- 5489, Rome, Italy FAO (2012) Aquaculture topics and activities. State of
world aquaculture. Text by Rohana Subasinghe. In: FAO Fisheries and Aquaculture
Department [online]. Rome. Updated 27 May 2005.
 FAO (2011). Fisheries and Aquaculture in Eastern Africa: An overview of the sectors
challenges and strategic priorities for development. SFE Technical Paper 52 pp.
 Jamu DM and Ayinla OA (2003) Potential for the development of aquaculture in Africa.
NAGA 26(3), 9-13, http://worldfishcenter.org/Naga/naga26_3.htm
 Reyntejens, D. and Tesfaye Wudneh (1998). Fisheries management – A review of the
current status and research needs in Ethiopia. SINET: Ethiop. J Sci.21:231-266.
 Watanabe, T. (2002) Strategies for further development of aquatic feeds. Fisheries
Science 68, 242 252.
 Yang, Y., Xie, S., Cui, Y., Zhu, X., Lei, W. & Yang, Y. (2006) Partial and total
replacement of fishmeal with poultry by-products meal in diets of gibel carp, Carassius
auratus gibelio Blouch. Aquaculture Research, 37, 40-48.
Activity 3: Evaluate culturing techniques of the freshwater rotifer, Brachionus calyciflorus

and its application in fish larviculture

The establishment of hatcheries and inducement of spawning by hormone injection greatly hold
promises for enhanced propagation of most fish species whose larvae can be maintained by
feeding with rotifers. Techniques that will be developed for all year round availability of most
freshwater species must co-opt the technology of feeding with the freshwater rotifer at least for
the first few days of larval culture to enhance its growth and chances of survival. Most fish
larvae are small in size and are very slow swimmers and so require rotifers. Rotifers are valuable
live food for the culture of the larvae of most fish species. Several characteristics of rotifers
including their very small size, relatively slow motility have contributed to their usefulness as
good prey for active larvae (Lubzens et al., 1989). In addition, they have the habit of staying
suspended in the water column, high reproductive rate and high density cultures. They can
tolerate temperatures of between 15 and 31 0C. The optimal pH is 6-8 at 25 0C (Ludwig, 1993).
Brachionus calyciflorus is of course the commonly cultured freshwater rotifer for both
freshwater fish species.
Since these early days, the demand for rotifers has continuously increased and several culture
techniques have been developed for rotifer mass production (Fukusho, 1983). Most popular
techniques in laboratory and commercial hatcheries are batch cultures, semi-continuous cultures
and the high density mass culture of rotifers on algae. Each culturing conditions have their own
merits and demerits.
Batch cultures
Batch cultivation, due to its simplicity, is probably the most common type of rotifer production
in marine fish hatcheries (Snell, 1991). The culture strategy will consists in either the
maintenance of a constant culture volume with an increasing rotifer density or the maintenance
of a constant rotifer density by increasing the culture volume. In the batch culture, a total harvest
of the rotifers is applied with part of the rotifers used as food for fish larvae and part used as
inoculums for the next culture (Hirata, 1980; Lubzens, 1987). Generally, the size of the tanks for
batch culture is flexible, ranging from 500- to 1000 liter plastic tanks up to as large as 10 tons
concrete tanks can be used. Culturing can be started from 200 to 250 rotifers ml -1. Many
disadvantages are, however, attributed to the batch culture system: the cultures are subjected to
highly variable conditions both in growth performance in terms of biochemical composition of
the rotifer population, unstable physico-chemical water parameters, low efficiency in terms of
labour and utilization of infrastructure. These problems contribute to unstable/unpredictable
culture conditions and a relatively low production yield (Walz et al., 1997) at high cost. A lot of
improvements have been made to create more stable culture and rotifer production in batch
cultures. Walz et al. (1997) have developed a turbidostat system. In this system, the rotifer
production is stabilized by maintaining constant algal densities by turbidity regulation.
There are considerable reports on feeding successes using the rotifer, Brachionus as live food for
several marine fish species (Villegas et al., 1990; Reitan et al., 1993; Ottera 1993, Craig et al.,
1994; Castell et al., 2003). However, limited information is available on culturing of aquatic
organisms and using as live food in fish larviculture in the tropics (Lim and Wong, 1997; Awaiss
and Kestemont, 1998). However, the use of rotifers would enable freshwater fish larviculturist to
improve larval performance and increase yield. High specific growth rate and survival in the
African catfish Clarias anguillaris larvae fed on rotifers (Mercier et al., 2004; Arimoro, 2004)
have been reported.
National Fishery and other Aquatic Life Research Center has started to produce quality fish
seeds in the hatchery. It is also known that the success in the hatchery production of fish
fingerlings for stocking in the grow-out production system is largely dependent on the
availability of suitable live food organisms such as rotifers and Artemia. However, the feed
particularly for the newly hatched fish fry is a bottle neck for the long awaited journey of the
research center. The absence of commercially prepared fish feed and some live foods like
Artemia in the country as well as a developed culturing technique are an opportunity for the
researchers to look alternative live foods and develop culturing techniques.
Objective (s):
 to adapt appropriate culturing techniques of Brachionus calyciflorus as live food for
larval fish
 to evaluate the growth and survival rate of Clarias gariepinus larvae fed on live foods
Material and methods
Food preparation for rotifers
To achieve high biomass production of rotifers researchers have to find alternative and cheap
food source. To achieve this, the culture medium will be fertilized with chicken manure and/or
fertilizers such as nitrates, phosphates for growth. Freshwater algae such as Chlorella and
Scenesdesmus are important useful algae for the culture of the freshwater rotifer. To develop the
required algal as food for cultured rotifer, pond water will be filtered using 40 µm mesh size
plankton net. The filtered pond water will be kept in aquaria with light access to allow algal
growth. Different algal species will grow in the aquaria and we will conduct repeated sub-
culturing technique to achieve the required algal culture.
Brachionus calyciflorus stock culture preparation

For starting and initiating the rotifer culture, mixed population of zooplankton comprising of
rotifers, copepods and cladocerans will be collected from the research ponds of National Fishery
and Other Aquatic Life Research Center (NFALRC) using 40 µm mesh size plankton net. The
freshwater rotifer B. calyciflorus will be isolated from other zooplankton groups and other rotifer
species by repeated sub-sampling from wild (pond sampled) stock. Stock cultures will be kept in
aquaria with light access which has selected algal culture.
Culture techniques
From the three rotifer culturing techniques, (batch cultures, semi-continuous cultures and the
high density mass culture) batch culture of rotifers on algae will be evaluated at room
temperature in the hatchery. Petridishes, glass jars and/or aquaria will be used to prepare culture
media for rotifers. Once the one or two algal species are cultured in the jars, the freshwater
rotifer, B. calyciflorus will be inoculated into the culture media. Rotifer culture with algae and
baker yeast will be compared to evaluate their impact in the production of the freshwater rotifer.
The amount of baker’s yeast fed on a daily basis will be at the rate of 1g million-1 of rotifers.
Application of rotifers to fish larviculture

Survival and growth rates of Clarias gariepinus fry fed with a rotifer, B. calyciflorus and
Artemia nauplii during the first week of larval feeding will be investigated. Application and rate
of rotifer feeding will be compared in the survival and growth experiment of the larval fish.
Location: NFALRC
Duration: 2 years
Work plan:
Activities 2009
Algal culture preparation X
Rotifer stock preparation X
Culturing Brachionus calyciflorus X
Running the experiment & larval feeding X
Data collection & data entry X
Data analysis X
Progress report X
Write up X
Budget break down
Budget codes 2009
6114 2000
6218 40,000
6219 65,000
6231 3000
Sub-Total 110,000
Contingency 11,000
Total 231,000
Expected outputs
• Batch culturing technique of rotifer adopted
• Guide line for larval fish live food feeding available
Responsibilities:
Center (s): NFALRC
Person (s): Adamneh Dagne, Kibru Teshome, Esayas Alemayehu

Monitoring and evaluation matrix
Activitytitle: Evaluate culturing techniques of the freshwater rotifer, Brachionus calyciflorus and
its application in fish larviculture
Output M&E Indicators Informatio Method of Tools for Method of

Objectives n to be collection collecting the analysis
collected information
Batch culturing To ensure No. of rotifer No. of eggs Microscopic Microscope, Microscopy,
technique of rotifers the production cycles, and hatched observation, thermometer statistical
adopted implementati pure mono algal rotifers, counting software
on of the cultures
activity & to
take
immediate
action/decisi
on
Activity: Algal culture preparation, rotifer stock preparation, culturing Brachionus calyciflorus, running the experiment & larval
feeding, data collection, entry and analysis
Inputs: Researchers, TAs, aquaria, outdoor tankers, water, algae, rotifer, budget
References
Arimoro FO, Ofojekwu PC (2004). Some aspects of the culture, population
dynamics and reproductive rates of the freshwater rotifer, B. calyciflorus fed
selected diets. J. Aquatic Sci. 19(2): 95-98.
Awaiss K. (1998). Feeding sequences (rotifer and dry chemical
composition of African catfish, (Clarias gariepinus) Burchell (Pisces:
Clariidae) Larvae. Aquaculture Res. 29(10): 731.
Castell J., Blair T., Neil S., Howes K., Mercer S., Reid Young-Lai J., Gullison B.,
Dhert P. and Sorgeloos P. (2003). The effect of different HUFA enrichment
emulsions on the nutritional value of rotifers (B. plicatilis) fed to larvae
haddock (Melanogrammus aeglefinus) Aquacult. Int.11(1–2): 109 – 117.
Coves D., Audineau P. and Nicolas JL. (1990). Rotifer-rearing technology. In:
Barnabe, G.(Ed.), Aquaculture, vol. I, Ellis Harwood, West Sussex, England,
pp. 232–245.
Craig SR, Connie R. and Holt JG. (1994).The effects of enriching live foods with
unsaturated fatty acids on the growth and fatty acids on the growth and fatty
acid composition of larval red drum Sciannops ocellatus. J. World
Aquacult.Soc. 25(3): 424-434.
Fu Y., Hada H., Yamashita T., Yoshida Y. and Hino, A. (1997). Development of
continuous culture system for stable mass production of the marine rotifer
Brachionus. Hydrobiologia 358, 145–151.
Fukusho K. (1983). Present status and problems in culture of the rotifer
Brachionus plicatilis for fry production of marine fishes in Japan. Symposium
International De Aquacultura, Coquimbo, Chile, Sept. pp. 361–374.
Hirata H. (1980). Culture methods of the marine rotifer, Brachionus plicatilis. Min.
Rev. Data file Fish. Res. Kagoshima Univ. 1, 27–46.
Hirayama K, Mariyama I. and Maeda T. (1989). Nutritional effect of freshwater
chlorella on the growth of the rotifer, Brachionus plicatilis. Hydrobiologia
186/187: 39-42.
Lim LC. and Wong CC. (1997). Use of the rotifer, Brachionus calyciflorus Pallas,
in freshwater ornamental fish larviculture Hydrobiologia 358 (1/3): 273 (5).
Lubzens E. (1987). Raising rotifers for use in aquaculture. Hydrobiologia 147,
245–255.
Lubzens E., Tandler A. and Minloff G. (1989). Rotifers as food in Aquaculture.
Hydrobiologia 186/187, 387– 400.
Ludwig GM. (1993). Effects of Trichlorfon, Fenthion, and Diflubenzuron on the
zooplankton community and on the production of the reciprocal-cross hybrid
stripped bass fry in culture ponds. Aquaculture 110: 301-319.
Mercier L., Audet C., Joel de la N., Parent B., Parish CC. and Ross NW. (2004).
First feeding of winter flounder (Pseudopleronectus americanus) larvae: use
of B. plicatilis acclimated at low temperature as live prey Aquaculture 229(229
(1-4):273-278.
Ottera H. (1993). Feeding, growth and survival of Atlantic cod (Gadus morhua)
Larval reared in replicate plastic enclosure. Can J. Fish Aquat. Sci. 50(5): 913 –
924.
Reitan L., Kjell T., Jose R., Gunvor O. and Olsen Y. (1993). Nutritional effects of
algal addition in the first feeding of turbot Scophthalmus maximus larvae.
Aquaculture 188(3): 257-275.
Snell, T.W., 1991. Improving the design of mass culture systems for the rotifer,
Brachionus plicatilis. In: Fulks, W., Main, K.L.(Eds). Rotifer and Microalgae
Culture Systems. Proceeding of US–Asia Workshop, Honolulu, Hawai, Jan. 28–
31, pp. 61–71.
Villegas CT., Millamena O. and Escritor F (1990): Food value of B plicatilis fed
three selected algal species as live food for milk fish Chanos chanos Forsskal
fry production. Aquacult. Fish. Mgt. 21: 213 – 219.
Walz, N., Hintze, T., Rusche, R., 1997. Algae and rotifer turbistats: studies on
stability of live feed cultures. Hydrobiologia 358, 127–132.
Yoshimura K., Iwata T., Tanaka K., Kitajima C. and Isizaki F. (1995). A high
density cultivation of rotifer in an acidified medium for reducing undissociated
ammonia. Nippon Suisan Gakkaishi 61, 602–607.
Yoshimura KA., Hagiwara A., Yoshimatsu T. and Kitajima C. (1996). Culture
technology of marine rotifers and implication for intensive culture of marine
fish in Japan. Mar. Freshwater Res. 47, 217–222.
Activity title 4: Effect of Incorporation of Fish Silage into Diets on Growth Performance
and Body Composition of Nile Tilapia (Oreochromis niloticus)fries
Background and Justification
Aquaculture is now the fastest growing food producing sub sector in many countries and it is
expected that this trend will continue despite numerous constraints, which may become more
challenging in the future(Matthias et al., 2001). In many developing countries there is significant
scope for enhancing contributions of aquaculture to food supplies and poverty alleviation (FAO
1999). However, according to FAO (1999) aquaculture in developing region is facing significant
challenges and among those meeting the growing demands for seed, feed and fertilizers, in terms
of quantities and quality have the highest priority.
Challenges for the development of aquaculture in Ethiopia include not only a lack of knowledge
and institutional support, but also a shortage of several production factors; among them the lack
of reasonably priced and locally available fish feed of sufficient nutritional quality.
Commercially produced compound feeds are readily available for aquaculture in developed
countries. In most developing countries formulated feeds for fish are scarce or entirely
unavailable (FAO, 2005). Although some developing countries are importing formulated fish
feeds, these are usually too expensive for an economically viable fish production. Since feed
costs represent 40-50% of the total variable production costs (Craig and Helfrich, 2002), locally
produced and reasonably priced feedstuffs of sufficient nutritional quality are a key element in
the development of aquaculture in countries like Ethiopia (Gabriel et al., 2007).
As aquaculture production becomes more and more intensive, fish feeds will be a significant
factor in increasing the productivity and profitability of the sector (Jamu & Ayinla, 2003).
Because feed management determines the viability of aquaculture as it accounts for at least 40 -
60 per cent of the cost of fish production (Shang 1992; Craig & Helfrich 2002; Jamu and Ayinla
2003).
Fishing operations yield includes considerable waste materials such as by-catch fish, un
marketed fish and filleting scrap. Unfortunately, the lack of processing facilities in many lesser
developed countries like Ethiopia often leads to the paradoxical situation of wastage accurring
where the need for protein is greater. Although priority should be given to the direct use of fish
for human consumption, there are often considerable quantities of fish wastes available for
animal feeding.
In Ethiopia, where supplies of fish waste may be small and irregular, it is uneconomical either to
transport the waste to a fish meal plant or to build a small local factory. Fish ensilaging may be a
suitable alternative method of preserving fish waste in these circumstances. Acid preserved fish
silage is a liquid product prepared by adding acid to fish or fish waste and liquefaction is caused
by proteolytic enzymes, present naturally in the fish (Backhoff, 1976). The addition of acid
stimulates enzymic proteolysis which help in dissolving bones and prevents bacterial and fungal
spoilage (Wingall and Tatterson, 1976). Feeding trials have shown that fish silage can replace
fish meal in the diets of salmon (Jackson et al.t 1984) and carp (Venujopal and Keshavanath,
1984).
Therefore the present study is aimed at investigating the effect of incorporating fish silage on the
growth performance and body composition of O.niloticus.
Objectives
General Objective
The general objective is to study the effect of incorporating fish silage into the diets of O.
niloticus on the growth performance.
Specific Objectives
 To adopt fish silage preparation technique

 To formulate and identify the best ratio of silage incorporated feed for O.niloticus
juveniles
Preparation of fish silage
To prepare fish silage, those fishes which are un-used for human consumption will be collected
from landing sites of lake Ziway and transported to Ziway Fishery and Aquatic Life Research
Center (ZFALRC) laboratory. Then before preparing the silage, the fish will be washed, minced
and homogenized. Then, the silage will be prepared according to Espeet.al., 1999. During fish
silage preparation, a 3% by weight solution of 98% formic acid will be added to the
homogenized fish mixture and mixed well. Then it will be stored for 48 days by homogenizing
once in a day. Before formulation of the experimental diets, fish silage will be neutralized by
adding 1.6% calcium hydroxide to raise the silage pH from 4.1 to 5.2. Then, fish silage will be
mixed with wheat bran to formulate other diets, containing, 0%, 25%, 50%, 75% and 100% fish
silage.
Formulation of Feed
The ingredients of the experimental diets will be ground and mixed well to pass through 1mm
pore sieve. Thereafter, the grounded feeds will be mixed homogeneously in the ratio of 0:100,
25:75, 50:50, 75:25 and 100:0 of S(silage):WF(wheat bran).
Experimental Unit
A total number of 180 fingerlings of Oreochrornis niloticus will be collected and grouped into
twelve glass aquaria containing equal amount of water. Each aquarium will be supplied with
compressed air through air pump for water aeration. Dechloronated tap water will be used for
replacing about one third of the total water volume in each aquarium daily after the removal of
fecal wastes. Water temperature will be controlled thermostatically by automatic heaters to
maintain in the range between 27-28 C°.
Growth Performance
The growth performance will be studied for a total of 16 weeks after start. During the
experimental period, fish will be fed the experimental diets at a rate of 3% of the live body
weight daily and the feed will be offered twice a day at 9.0 a.m and 3.0 p.m. The fish groups will
be weighed weekly and the amount of the feed will be adjusted according to the actual body
weight changes. Water samples will be taken each week from each aquarium to determine water
quality parameters. One third of water volume will be changed daily and the whole water volume
will be totally changed every week.
Growth and feed utilization parameters
Average weight gain (AWG), average daily gain (ADG), specific growth rate (SGR % d),
feed/gain ratio, feed conversion ration (FCR), protein efficiency ratio (PER) and survival rate
will be calculated according to the following equations:
a) Average weight gain (g/fish) (AWG):
WG = W2-W1
Where: Wl = the initial weight (g)
W2 = the final weight (g)
b) Average daily gain (ADG)
Average daily gain (ADG) will be estimated according to the following formula
ADG = W2- Wl
Where: Wl = The initial weight (g)
W2 = The final weight (g)
T = Experimental period (d)
c) Specific growth rate (SGR)
Specific growth rate (SGR) will be estimated according to the following equation:
SGR =Ln.W2-Ln.Wlxl00
Period (days)
Where: Ln = Natural Logarithm (log)-10
Wl = Mean initial weight (g)
W2 = Mean final weight (g)
d) Relative growth rate (RGR)
The relative growth rate will be calculated according to the following equation
RGR = W2- Wl x lOO
Wl
Where: Wl = The initial weight
W2 = The final weight
e) Feed conversion ratio (FCR)
FCR = Dry feed intake (g)
Live weight gain (g)
f) Protein efficiency ratio (PER)
PER = Live weight gain (g)
Protein intake (g)
Proximate analysis
Proximate analysis for silage, experimental diets, and whole fish bodies will be carried out for
moisture, ash, crude protein and fat according to the methods described by A.O.A.C. (1990).
Water quality parameters
Samples of water will be taken weekly from each aquarium for determination of water
temperature using a water thermometer (daily), water pH value using digital pH meter, dissolved
oxygen concentration using an oxygen meter. Analyses of NO 2, NO3, alkalinity and Hardness
will be carried out using kits.
Statistical analysis
The numerical data will be statistically analyzed using SPSS package for one-way analysis of
variance. When F-test result will be significant, least significant difference will be calculated
according to Duncan multiple range test.
Location
The study will be conducted in hatchery laboratory of National Fishery and Aquatic Life
Research Center, Sebeta, Ethiopia
Duration
3 years (2008-2010 E.C)

Work Plan
No. Title and detail activities Unit Month
J A S O N D J F M A M J
1 Laboratory analysis No. of

measurements
2 Growth performance No. of

evaluation measurements
3 Data analysis No. of days

Budget by code
Budget Code 2009E.C.
6114 15050
6212 5808.9
6213 2112
6215 5703
6217 7393
6218 10350
6219 8238
6223 5281
6231 45943
6232 2112
6241 11090
6243 3697
6244 9294
6245 13202
6256 2640
6271 16899
6313 19539
Total 184,352
Output
 fish silage preparation technique will be adopted
 best combination of silage based feed for O. niloticus will be identified
Responsibilities
Bezuayehu Gutema, Abelneh Yimer, Esayas Alemayehu, Emebet Kebede,

Tools for
M & E Information to Method of collecting the Method of
Out put Objectives Indicators be collected data collection information analysis
-best ratio of - to ensure -average weight -weight of fish - sampling and -hand nets -statistical
fish silage the activity gain weighing of software
-water quality -weighing
based feed is carried out fish
-average daily gain parameters balance
successfully
-sampling and
as per the -specific growth -weight of feed -
analysis of
plan rate intake
water quality
-relative growth parameters in
rate laboratory
-feed conversion
ratio
-protein efficiency
ratio
Activities
- Feed - to ensure
formulation, proper
setting up accomplishm
experimenta ent of sub
l area activities
(aquarium), following
fish feeding, technical
water
quality
monitoring
and growth
data
monitoring
Inputs
-
References
 Craig, S., & Helfrich, L. A. (2002). Understanding fish nutrition, feeds, and
feeding. Virginia cooperative extension, 63, 256-270.
 Halwart, M., Funge-Smith, S., & Moehl, J. (2003). The role of aquaculture in rural
development. Review of the State of World Aquaculture, FAO, Rome, 47-58.
 Tacon, A. G., & Nates, S. F. (2007, May). Meeting the feed supply challenges of
aquaculture. In Global trade conference on aquaculture. FAO Fish. Proc (Vol. 9, pp.
117-121).
 Jamu, D. M., & Ayinla, O. A. (2003). Potential for the development of aquaculture in
Africa. NAGA, WorldFish Center Quarterly, 26(3), 9-13.
 Kassahun, A., Waidbacher, H., & Zollitsch, W. (2012). Proximate composition of
selected potential feedstuffs for small-scale aquaculture in Ethiopia. Livestock Research
for Rural Development, 24(6).
 Craig, S., & Helfrich, L. A. (2002). Understanding fish nutrition, feeds, and
feeding. Virginia cooperative extension, 63, 256-270.
 BACKHOFF, H. P. (1976). Some chemical changes in fish silage. International Journal
of Food Science & Technology, 11(4), 353-363.
 Wignall, J., & Tatterson, I. (1976). Fish silage. Process Biochemistry, 11(10), 17-19.
 Venugopal, M. N., & Keshavanath, P. (1984). Influence of supplementary feeds on the
biochemical composition of flesh of freshwater carps Catla catla (Ham.) Cirrhinus
mrigala (Ham.) and Cyprinus carpio (Linn.). Indian Journal of Animal Sciences.
Activity 5: Comparative assessment of proximate composition and mineral content in
farmed and wild Nile Tilapia (Oreochromis niloticus)

Fish provides superior quality protein to that of meat, milk and eggs and well balanced essential
amino acid profile, necessary minerals and fatty acids. In addition to that fish flesh is tasty and
highly digestible. Over and above it minimizes the risk of heart diseases and increases life
expectancy.
Protein, fat, ash and moisture (proximate composition) are traditionally used as indicators of the
nutritional value of fish. Nile Tilapia as an important food fish in developing counties is widely
used in commercial farming systems for intensive aquaculture. Fish feeds supplement were
critical in intensive system, and is usually a major part of production cost. Therefore, fish
nutritionists have made several attempts to partially or totally replace animal derived protein
with less expensive, unconventional and locally available plant protein source. However, plant
protein source are full of anti-nutritional factors which reduces bioavailability of nutrients.
Therefore it is imperative to compare the nutritional composition of Nile Tilapia fillets obtained
from wild and culture.
Objective
 To compare proximate composition and mineral content in farmed and wild Nile Tilapia
Description of the study area
The Nile Tilapia culturing will be conducted at Arsinegelle district of West Arsi zone. Data to be
taken during pond culture are type of commercial feed used, feeding time and rate, stocking
density of fish in the pond, average water temperature, salinity and pH.
Proximate composition of Experimental Feeds

The proximate composition (moisture content, crude protein, crude fat and ash) of experimental
feeds will be analyzed according AOAC procedure.
Bioavailability of experimental feeds
To calculate bioavailability of minerals in experimental feeds, anti-nutritional factors like

phytate, oxlate and tannin will be determined at Ethiopian Health and Nutrition Research
Institute.
Sample collection and preparation
Table size adult farmed (cultured) Nile Tilapia will be obtained from Aquaculture farm which is
cultured for other purposes by our center.Approximately equal weight to farmed fish will be
taken from the same water bodies used as fingerlings source for fish culturing. Live fish will be
transported to Zeway fisheries resources research center layering with flaked ice using ice box.
Fish specimen will be cleaned, descaled, eviscerated and filleted manually using plastic knife.
Proximate composition determination
Moisture content
Immediately after filleting, moisture content will be determined using oven drying methods. For
the purpose 5 g of fillet will be placed in oven and dried at 105 0c for 3 hours until constant
weight is obtained. The remaining fillet will be dried at 60 0 c for 72 hours and then dried fillets
will be finely powdered using laboratory mill to 0.3mm size. Finally the powder will be packed
and stored in desiccators for further analysis.
Weigh tofwetsample−weig h tofdriedsample

Moisturecontetnt= ×100
Weig htofwetsample
Determination of crude protein

Crude Protein in fish fillets will be determined by Kjeldahl methods. 0.5 g of powdered fish fillet
will be weighed into Kjeldahl flask and will be digested by heating at 370 0c for four hours in the
presence of 6 ml sulfuric acid (H2SO4), 3.5 ml Hydrogen peroxide (H2O2) and 3 g of catalyst
copper sulfate (CuSO4) and potassium sulfate (K2SO4). After digestion will be completed,
formed clear solution will be cooled for 30 minutes and neutralized by addition of 25 ml of
NaOH (40 %) and diluted using 25 ml distilled water. Then 25 ml of distilled water, 25 ml of
boric acid and 3 drops of methyl blue will be added into receiving flask of 250 ml capacity
connected to the distiller by tube. The distillation process will be terminated when the volume of
receiving flask reached between 200 to 250 ml. Eventually the nitrogen content will be estimated
by titration of the borate anion with 0.1 N HCl using the following formula.
( VolofHCl consumed bysample−VolofHClconsumedbyblank ) L g

% N =N HCl× X 14 X 100
Weig htofsample mole
Note: all reagents will be added to blank except the sample.
Determination of crude fat
Crude fat will be determined by semi-continuous solvent extraction methods (Soxhlet method).
Accordingly 2 g of fine powdered fish fillet will be placed in porous cellulose extraction thimble
and covered with fat free cotton. The thimble will be placed in extraction chamber which is
suspended above a flask containing 50 ml diethyl ether. The flask will be heated at 55 0 c and the
solvent evaporates and moves up into the condenser where it converted into a liquid that trickles
into the extraction containing the sample. At the end of extraction process, which typically lasts
for 3 hours, the flask containing the solvent and lipid will be removed, the solvent will
evaporates at 70 0c and the weight of lipid remaining will be quantified gravimetrically.
Weig h toffat
Fatcontent = X 100
Weig h tofsample
Determination of total ash
Total ash content will be determined using dry ashing method. For the purpose, 2 gram of
powdered fish fillet will be weighed into ashing crucibles, placed on a hot plate under a fume
hood and the temperature will slowly increased and awaited until smoking ceases and the
samples become thoroughly charred. The crucibles will be placed inside muffle furnace set at
550oc for 4 hours, and removed from the muffle furnace and then placed in desiccators for 1 hour
to cool. The amount of ash in the sample will be measured from difference in weights and
expressed as
Massofcrucibleswit h as h−Massofemptycrucibles
As h content= X 100
Massofsample
Calculation of gross energy
Gross energy value (Kcal/ 100g) will be calculated according to Atwater’s conversion factors; by
overall addition of the protein content multiplied by 4 and total fat content by 9.
Minerals analysis
The ash obtained from wet ashing will be wetted completely with 5 ml of 6 N HCl and dried on a
low temperature hot plate. 7 ml of 3N HCl will be again added to the dried ash and heated on the
hot plate until the solution just boils. The will be cooled to room temperature in a hood and
filtered into 50 ml volumetric flask using a 125 mm filter paper. The cooled solution will be
filtered through Whatman filter paper No. 41 and made up to 25ml with 2N nitric acid; a ``blank''
will be prepared in a similar manner. Phosphorus will be determined by UV-VIS
spectrophotometer at 715nm using the ammonium Phosphomolybdate method. Sodium and
potassium will be determined by flame photometer (AOAC, 969, 23) while Calcium, Iron,
Magnesium and Zinc will be determined by Flame Atomic Absorption Spectrometry.
Data management
All proximate composition and minerals content will be done in triplicate. Proximate
composition and minerals data will be from both wild and culture will be compared using student
t-test.
Work plan
Months
Activities J A S O N D J F M A M J
Laboratory arrangement X
Purchase of chemicals and equipments X
Sample collection X X X
Sample analysis X X X
Data coding, entry and analysis X X X
Write up X
Responsibles: Alemu Lemaa, Getacew Senbete and Bezuayehu Gutema
Budget by code
Budget Items Unit price Quantity Total price

code
6114 20000
6216 10000
6232 50000
11760
Polyethylene bags 0.5 168 84
Copper Sulfate (CuSO4) 650 1kg 1600
Potassium Sulfate (K2SO4) 650 1kg 650
Conc. Sulfuric acid (H2SO4) 520 1L 3000
Hydrogen peroxide (H2O2) 150 1L 1500
Hydrochloric acid (HCl) 450 1L 2000
Diethyl ether 300 10L 13000
Thimble 3000 1 pack 6000
Sodium Hydroxide (NaOH) 220 2kg 8000
Methyl blue 240 0.25L 60
Cotton wool 45 1 roll 45
Boric acid (H3BO3) 320 1kg 3000
Boiling chips 0.5 50 pcs 25
Total 18562
References
LFDP. 1997. Lake Fisheries Development Project, Phase II. Working paper number twenty
three. Second edition. Lake management plan. Fisheries Resources Development
Division (MoA).
Association of Official Analytical chemists (AOAC). 1998. Official methods of Analysis of
AOAC international. 16th Edition. 4th revision.
Activity 6: Laboratory scale Isolation and Culturing of Spirulina (Arthrospira) using

standard medium supplemented with soda lake water
Background and Justification
Arthrospira commercially known as spirulina is a blue green multi cellular filamentous micro
algae that have a commercial importance. It is used as a source of protein (50–70 % protein by
dry weight, exceeding all known standard plant proteins), unusual fatty acids (gamma-linolenic
acid), vitamins (e.g., β- carotene and B12), minerals (e.g., iron) and essential amino acids. It has
also healing effects on various health problems including hyperlipidemia, nephrotoxicity,
diabetes, obesity, hypertension, cancer and even HIV (Belay and Ota 1993; Belay et al. 1996;
Belay 2002).
Arthrospira has gained considerable popularity in the health food industry and increasingly as a
protein and vitamin supplement to aquaculture diets (FAO, 2008). As a result, commercial scale
production of Arthrospira has got global attention. It is one of the microalgae that reproduce fast
and has high productivity (Kilic et al. 2006). A number of environmental factors including
carbonate–bicarbonate alkalinity, pH, nutrient availability, temperature and light contribute for
the growth and biomass production of Arthrospira (Vonshak 1997; Habib et al. 2008). Biomass
of Arthrospira can be increased when these environmental factors are optimized. It preferably
dominates in tropical and sub-tropical water bodies characterized by high carbonate–bicarbonate
alkalinity and pH (Ciferri 1983; Cogne et al. 2001). However, optimization of growth conditions
is the major challenge to biomass production of Arthrospira in cultivation systems and it remains
coasty. Nutrients account 15-25% of the total production cost of the total production cost of
Arthrospira biomass, which form the second major cost item (Belay 1997; Vonshak 1997; Habib
et al. 2008). Therefore, standard carbonate medium is unaffordable for production of Arthrospira
biomass production.
Various biotechnological researches and innovations were undertaken to investigate low cost,
alternative methods of cultivation without reducing biomass productivity (Vonshak 1997).
Promising results have been found in experiments using seawater enriched with phosphate and
urea. In such experiments, Materassi et al. (1984) and Tredici et al. (1986), in laboratory and
outdoor mass cultivations, respectively, obtained biomass yields which are slightly less than
those obtained with the standard medium. Studies in Brazil by Costa et al. (2003) have also
shown that comparable Arthrospira biomass could be obtained using lagoon water supplemented
with bicarbonate and urea. Use of different organic sources of carbon, nitrogen and phosphorus
has also resulted in increased biomass yield of Arthrospira (Neilson and Larsson 1980; Baldia et
al. 1991; Habib et al. 2008). Although soda lakes are conducive for Arthrospira, supporting its
abundant population, there are limited studies to evaluate their potential for biomass production
under laboratory and outdoor conditions.
Ethiopia has alkaline soda lakes such as lakes Chitu and Shala, which have conducive features
for Arthrospira (Tadesse et al., 2014). Lake Chitu is a small shallow lake best known for its
natural monoculture of Arthrospira (Talling et al. 1973; Kebede 1996). Harvesting Arthrospira
biomass from the natural ecosystem of the small lakes like Chitu will not be sustainable
ecologically and economically (Tadesse et al., 2014). Lake Shala is a large and deep soda lake
whose gross water chemistry is very similar to that of the nearby Lake Chitu and which is
suitable for Arthrospira production (Wood and Talling 1988; Kebede 1996). Location of the
lakes is in areas with high temperature and irradiance, and nearly constant photoperiod, which
seem to be ideal tropical climatic conditions supporting the high productivity of Arthrospira
(Richmond and Grobbelaar 1986; Vonshak 1997; Talling and Lemoalle 1998).
Evaluation of growth and biomass production of Arthrospira in waters of the soda lakes Chitu
and Shala with or without supplementation with standard Spirulina medium (Zarrouk medium as
modified by Aiba and Ogawa (1977) ) has been conducted and comparable growth and biomass
production were achieved (Tadesse et al., 2014). The result also reveals the need for further
studies to optimize the medium proportions particularly in fulfilling nitrogen requirement of
Arthrospira for better growth and biomass production. Therefore, this study aimed to identify
alternative recommendations of Chitu and Shala lakes water with and without supplementation
of different standard Spirulina medium (CFTRI medium) for enhanced growth and biomass
production.
Objectives
 To adopt Arthrospira culturing technique in a laboratory scale

 To identify the best combination of standard medium (CFTRI) and soda lake water for
enhanced productivity of Arthrospira.
Materials and Methods
Measurement of Physicochemical Parameters of the Soda Lakes
In situ measurement of pH, temperature, salinity and conductivity will be done before collection
of the biological material and 10 liter water samples will be collected from each soda lake for
physicochemical analysis of some parameters in laboratory. Alkalinity will be determined by
titration with 1N HCL to pH 4.5 using a mixed indicator (Bromocrysol green-methyl red).
Carbonate-bicarbonate alkalinities as CaCO3 and their ions will be calculated from total
alkalinity and pH according to APHA et al. (1999). NO 3−, NH3, PO4-P and SO4 2−
will be
determined, using samples filtered through Whatman GF/F, by sodium salicylate (APHA 1995),
phenate, ascorbic acid and turbidimetric methods, respectively (APHA et al. 1999). Samples
acidified to a pH of 2 with HNO3, will be used for the analysis of major ions and some
micronutrients according to the standard analytical methods outlined in APHA et al. (1999): Na+
and K+ by flame photometric method, Ca2+ by direct nitrous oxide-acetylene flame method, Cl−
by argentometric method, Mg2+, Fe, Zn, Mn, Cu, and Co by direct air-acetylene flame method.
Boron (B) was determined by azomethine H-colorimetric method (FAO 2008).
Sample Collection and Isolation
Water samples will be collected from lake Chitu and Shala using plankton net. A sub sample will
be taken and concentrated by centrifugation at 7000 rpm for 15 minute. The sediment will be
transferred to small test tubes containing standard medium and brought to laboratory for
isolation. Arthrospira trichomes will be isolated by serial dilution technique according to
Andersen and Kawachi 2005. An aliquot from a sample of the lake water will be diluted with
liquid SM, CFTRI in a small test tube from which drops will be transferred to multi-well plate
using Pasteur micropipette and concentration of trichomes will be checked under an inverted
microscope. After a series of similar dilutions and observations, some trichomes will be picked
up with the micropipette and introduced into 6 small test tubes of 15 ml capacity containing
about 4 ml of SM. The trichomes in the test tubes will be allowed to grow at a photon flux
density of about 20 μmol photons m−2 s−1 (produced by two fluorescent lamps, 36 W each) and
temperatures of 22–24 °C. Dense cultures of the micro alga will be diluted by the addition of the
SM and scaled up to a large volume (250 mL). Manual mixing of cultures will be applied four to
five times a day. These cultures will be used as a source of inocula for the experiment.
Preparation of Growth Media /Experimental Design/
Water from soda Lakes will be filtered through a plankton net (64 μm pore size) to remove large
microorganisms and particulate materials, within a few hours of its collection. Then, the filtered
water will be sterilized and neutralized chemically according to Kawachi and Noel (2005)
procedure. To evaluate growth responses of Arthrospira, with the aim to reduce the cost of
growth media by substituting standard medium with soda lake waters (by10%, 40 %, 70% or 100
%), nine different types of media will be prepared as indicated in the table 1 below.
Table 1: SM: Standard Medium, SLW: Saline Lake Water, CBW: Chitu Based Water, SBW:
Shala Based Water
Treatments Growth Medias R1 R2
Control SM:SLW (100:0)
T1 SM:CBW (70:30)
T2 SM:CBW (40:60)
T3 SM:CBW (10:90)
T4 SM:CBW (0:100)
T5 SM:SBW (70:30)
T6 SM:SBW (40:60)
T7 SM:SBW (10:90)
T8 SM:SBW (0:100)
The Experiment
The experiment will be carried out in 250 mL Erlenmeyer flasks holding 150 mL culture
medium in duplicate. The culture flasks will be inoculated with 15-mL aliquots, constituting 10
% of the final culture volume, that will be taken from exponentially growing cultures. The
cultures will be provided with an artificial light source from fluorescent lamps (three, 36 W
each), providing a photon flux density of about 50 μmol photons m −2 s−1 on the surface of the
culture. The temperature of the experimental cultures will be maintained using heater to regulate
the temperature of the bath at 35 °C and consequently to ensure constant temperature among the
treatments in a light–dark cycle of 10:14 for 18 days. These light and temperature conditions
were reported in several research articles as the optimal levels for Arthrospira growth in the
laboratory (Vonshak 1997; Oliveira et al. 1999; Andersen and Kawachi 2005). Mixing of the
cultures will be carried out manually by gentle shaking of the culture flasks four times a day. The
independent variable to be optimized will be the growth medium, keeping such growth
conditions as temperature, light and mixing constant. The response variables analyzed will be
specific growth rate (μ), biomass production (B) and doubling time (dt).
Laboratory Analysis
Before inoculation, initial value of the biomass index - chlorophyll-a (chl-a) of the inocula and
the initial pH and conductivity of all culture media will be measured. During the experimental
period, similar measurements will be taken every 2 days except for the first set of measurements,
which will be taken after 24 h to check the occurrence of a lag phase. Chl-a will be determined
spectrophotometrically from 5-mL samples filtered onto GF/F and extracted in 90%acetone and
the concentration of chl-a will be estimated using the equation in Vonshak (1997). Biomass
production (B, mg L−1) will be calculated as the change in biomass per volume of sample
filtered (Colla et al. 2007). Specific growth rate (μ) and doubling time (dt) will be calculated
using the equations used for batch culture of microalgae in the exponential growth phase
according to Guillard 1973; Vonshak 1997.
Statistical Analysis
The differences in growth parameters and some growth conditions of the various media will be
analyzed by one-way ANOVA, and Tukey’s LSD post-hoc test will be used for multiple
comparisons. The variables, which contributed to the observed variations in growth and biomass
production of Arthrospira in various media, will be determined by multiple regression analysis.
All statistical analyses will be done using SPSS statistical program.
Location
Samples will be collected from lake Chitu and Shalla, and the study will be conducted in
National Fishery and Aquatic Life Research Center, Sebeta, Ethiopia.
Duration
Two years (2009 to 2010 E.C)

Work plan
No. Title and detail Unit Annual plan Month

activities
J A S O N D J F M A M J
1 Laboratory medias and No of 5 growth medias *

experimental setup experime with full
preparation ntal experimental setup
medias
2 Isolation and culturing No. of 5 (water sample * *

of Arthrospira days collection)+
fusiformis 15(isolation)+ 30
(culturing)=
3 Water quality No. of 1(water quality )* * *

monitoring measurm 15 (times)= 15
ents
4 Proximate analysis No. of 1 *

measurm
ents
Budget by code
Budget Code 2009E.C
6114 23000
6212 3000
6213 85000
6215 20000
6217 36000
6223 46000
6231 32000
Total 245000
Output
 laboratory scale culturing technique of Artrospira fusiformis will be adopted

 appropriate ratio of SM with soda lake water will be identified for low cost Artrospira
production
Responsibilities
Bezuayehu Gutema, Kibru Teshome and Adamneh Dagne

Tools for
M & E Information to be Method of data collecting the
Out put Objectives Indicators collected collection information Method of analysis
-proper ratio of - to get low -Biomass -chlorophyll-a -Laboratory -water filtering -statistical software
SM and SLW cost medium production -water quality analysis apparatus,
for comparable for production -specific growth parameters (pH and centrifuge,
production of of Artrospira rate conductivity) spectrophotometer
Artrospira -doubling time
Activities
- experimental
setup
preparation
-strain
collection,
-isolation and
culturing of
the strain
-
Inputs
-
References
 Aiba S, Ogawa T (1977) Assessment of growth yield of a blue-green alga Spirulina

platensis in axenic and continuous culture. J Gen Microbial 102:179–182
 American Public Health Association (APHA) (1995) Standard methods for the
examination of water and wastewater, 19th edn. Washington, DC
 American Public Health Association (APHA), American Water Works Association
(AWWA), Water Environment Federation (WEF) (1999) Standard methods for the
examination of water 20th and. Washington, DC
 Andersen RA, Kawachi M (2005) Traditional microalgae isolation techniques. In
Andersen RA (ed) Algal culturing techniques. Elsevier, London, pp 83–100
 Ayala F, Vargas T, Cardenas A (1988) Chilean experiences on microalgae culture. In:
Stadler T,Mollion J, VerdusMC, Karamanos Y,MorvanH, Christiasen D (eds) Algal
biotechnology, proceedings of the 4th international meeting of the SAA. Elsevier,
London, pp 229–236
 Baldia SF, Nishijima T, Hata Y (1991) Effects of physicochemical factors and nutrients
on the growth of Spirulina platensis isolated from Lake Kojima Japan. Nippon Suisan
Gakkaishi 57:481–490
 Belay A (1997) Mass culture of Spirulina outdoors—the Earthrise Farms experience. In:
Vonshak A (ed) Spirulina platensis (Arthrospira): physiology cell-biology and
biotechnology. Taylor & Francis, London, pp 131–158
 Belay A (2002) The potential application of Spirulina (Arthrospira) as a nutritional and
therapeutic supplement. JANA 5:27–48
 Belay A, Ota Y (1993) Current knowledge on potential health benefits of Spirulina. J
Appl Phycol 5:235–241
 Belay A, Kato T, Ota Y (1996) Spirulina (Arthrospira): potential application as an animal
feed supplement. J Appl Phycol 8:303–311
 Tadesse O, Demeke K, Tadesse F, Baye F (2014) Evaluation of growth and biomass
production of Arthrospira (Spirulina) fusiformis in laboratory cultures using waters from
the Ethiopian soda lakes Chitu and Shala. J Appl Phycol 14: 251-254
 Chen F, Zhang Y, Guo S (1996) Growth and phycocyanin formation of Spirulina
platensis in photoheterotrophic culture. Biotechnol Lett 18:603–608
 Ciferri O (1983) Spirulina, the edible micro-organism.Microbiol Rev 47: 551–578
 Cogne G, Lasseur C, Cornet JF, Dussap CG, Gros JB (2001) Growth physiology of
microorganism (S. platensis) by pressure measurement. Biotechnol Lett 23:1309–1314
 Colla LM, Reinehr CO, Reichert CJ, Costa AV (2007) Production of biomass and
nutraceutical compounds by Spirulina platensis under different temperature and nitrogen
regimes. Bioresour Technol 98: 1489–1493
 Food and Agriculture Organization of the United Nations (FAO) (2008) Guide to
laboratory establishment for plant nutrient analysis. FAO Fertil Plant Nutr Bull 19, Rome
 Guillard RRL (1973) Culture methods and growth measurements division rates. In: Stein
JR (ed) Handbook of phycological methods. Cambridge University Press, Cambridge, pp
289–311
 Habib MAB, Parvin M, Huntington TC, Hasan MR (2008) A review on culture
production and use of Spirulina as food for humans and feeds for domestic animals and
fish. FAO Fisheries and Aquaculture Circular No 1034, Rome, pp 33
 Jourdan JP (1993) Solarium Spirulina farm in the Atacama desert (North Chile) In:
Doumenge F, Durand-Chastel H, Toulemont A (eds) Spiruline algue de vie. Bull Inst
Oceanogr Monaco Mus Oceanogr 12:191–194
 Kawachi M, Noel MH (2005) Sterilization and sterile techniques. In: Andersen RA (ed)
Algal culturing techniques. Elsevier, London, pp 65–81
 Kebede E (1996) Phytoplankton in a salinity–alkalinity series of lakes in the Ethiopian
rift valley. Dissertation, Uppsala University
 Kebede E (1997) Response of Spirulina platensis (Arthrospira fusiformis) from Lake
Chitu Ethiopia to salinity stress from sodium salts. J Appl Phycol 9:551–558
 Kilic C, Goksan T, Ak I, Gokpinar S (2006) Iki farkli Spirulina paltensis supunun
buyume ozelliklerinin karsilastirilmasi. EU J Fish Aquat Sci 23:189–192
 Legesse D, Gasse F, Radakovitch O, Vallet-Coulomb C, Bonnefille R, Verschuren D,
Gibert E, Barker P (2002) Environmental changes in a tropical lake (Lake Abiyata;
Ethiopia) during recent centuries. Palaeogeogr Palaeoclimatol Palaeoecol 187:233–258
 Neilson AH, Larsson T (1980) The utilization of organic nitrogen for growth of algae:
physiological aspects. Physiol Plant 48:542–553
 Tudorancea C, Zinabu GM, Elias D (1999) Limnology in Ethiopia. In: Wetzel RG, Gopal
B (eds) Limnology in developing countries, vol. 2.
 International Association for Limnology (SIL) International Scientific Publications, New
Delhi, pp 63–118
 UNESCO (1983) Algorithms for computation of fundamental properties of seawater.
 UNESCO technical papers in marine science 44, 58 pp
 Zinabu GM (2002) The effect of wet and dry seasons on the concentrations of solutes and
phytoplankton biomass in seven Ethiopian rift valley lakes. Limnologica 32:169–179
Activity 7: Comparison of periphyton and conventional formulated feed-based fish farming in

growth performance of Oreochromis niloticus in ponds
Responsibiles: Kibru, Abelneh, Fikadu-T, Fikadu-HM
Introduction
Periphyton is an community consisting of various taxa attached on submerged or floating substrates. Hunt
(1952) described periphyton to comprise sedentary organisms attached to submerged objects forming
slimy matrix and free living organisms associated with them (biofilms). Due to their large biomass
compared to phytoplankton, studies suggest that periphyton, together with macrophytes are positioned as
the main primary producers in ponds and shallow lakes (Saikia and Das, 2009).
Due to the growing understanding of periphyton biology, their functional role in the aquatic food web is
being established; they are already being used as fish food in fresh water aquaculture in many Asian
countries (Saikia and Das, 2009). Several studies indicated that periphyton serves as high quality food for
consumers including fish, which respond with increased assimilation efficiency and growth rates (Dugan,
2001). Due to economic reasons associated with modern aquaculture using formulated feed as the major
input that incurs high cost, fisheries scientists have frequently raised economic sustainability of those
systems(Azim et al, 2002; 2004b). As periphyton increases the total primary production of shallow
ponds, their potential as fish food for future aquaculture development is promising.
Small pond-based aquaculture is a recent effort to introduce aquaculture to local communities so that it
complements the existing farming tradition in which the most common pond inputs are wheat bran, mill
dust and other household cereal and house-hold food leftovers. Nevertheless, those inputs are increasingly
facing price hikes because these ingredients are used for livestock feeds, a sector that has deep rooted
tradition. Lack of affordable formulated feed with high quality therefore jeopardizes the advancement of
aquaculture in addition to other economic challenges. Therefore alternative production schemes are most
welcome.
In semi-intensive farming, periphyton can replace formulated diets thereby reducing feed cost to small
holder fish farmers which otherwise would contribute up to 60 percent of the total cost (El-Sayed, 1998).
For instance, biological and economic comparison of polyculture systems in a periphyton-based
aquaculture in Bangladesh revealed increased fish yield (up to 59%) over fertilized (with chemical
feritilizer) ponds indicating that periphyton based aquaculture can be profitable (Azim et al, 2004b).
Tilapia zilli and Cyprinus carpio are well known benthic omnivores. Saikia and Das (2008) found a
positive correlation between periphyton biomass and carp gut content and body condition in rice field
integrated fish farming with preference of periphyton to phytoplankton as the rice stem contained more
periphyton. Therefore, ecological fish farming technologies bring noble opportunities to the future of
freshwater aquaculture.
The price of formulated diets with all its limitations is becoming unaffordable for semi-intensive small-
scale fish farming. Alternative live feeds such as periphyton should be sought in order to agument the
economic challenge faced by the small-holder fish farmers. The aim of this study is to evaluate
periphyton-based tilapia farming and compare with that of conventional feed-based farming. The study
includes the bio-economic evaluation of both farming systems in terms of fish biomass and pond
management.
Objectives:
General objectives:
 To evaluate the possibility of substituting formulated tilapia feed with periphyton
Specific objectives
 To compare the growth performance of Nile Tilapia (O. niloticus) in ponds on-station
 To compare pond water quality dynamics in periphyton-based and feed-based tilapia farming
systems
Methods
Hypotheses:
• Improved water quality parameters will be achieved in periphyton-based ponds compared to feed-
based ponds
• Periphyton will result in comparable fish growth and survival rate as in feed ponds
The study involves a series of experiments in mainly two phases. The first phase of the experiment
comprises periphyton production optimization. Potential substrate materials on which periphyton
community biomass accumulates are evaluated using theoretical models developed previously by Degefu
et al (2011). Water quality parameters are also monitored over time and space along with periphyton
standing biomass. The second phase of the experiment comprises evaluation of periphyton based-fish
farming. Six ponds will be stocked with O. niloticus fingerling with average total length of 10cm at a
stocking density of 4 fish/m2. The ponds are grouped in to three treatment groups. Periphyton-based only,
periphyton combined with feed and feed only shall make up the three treatment groups. Fish will be fed
up to 5% body weight once daily for treatments involving feed supplementation. Fish growth rate
(specific and daily growth rate) are monitored every month by taking length-weight data of up to 20 fish
per pond. Water quality parameters are also monitored over the entire 8 month growth experiment
period. For the last three month of growth period, gut content will be collected from sampled fish from
all ponds for feed composition and relative importance. Water quality parameters such as dissolved
oxygen, temperature, pH, plankton biomass shall be monitored. Inorganic nutrients, total alkalinity, total
hardness are also monitored measured for both water source and pond water. Substrates for periphyton
treatments shall be planted up to a total surface area of 1m 2 per m2 of pond area.
Materials
• eight 5 x 8 m earthen/lined ponds
• Water source
• Substrate plant material – Giant cane - (Arundo donax), Arundinaria sp., and other wood
materials
• O. niloticus fingerling 10cm TL
• Giant cane stems of total surface area of 1m/m2 will be planted in T2 and T3
• Known proximate composition
• Laboratory facility and measuring equipments (e.g. probes)
Data handling
Raw data are processed in spreadsheet software. Water quality and fish growth measurement dynamics
are presented graphically and interpreted in-terms of threshold values. ANOVA will be used to determine
differences in the measurements between treatment groups (Tukey-test). The latter will also be used to
determine differences in fish growth performance (SGR,DGR, CF).
Location:
NFALRC Ponds
Duration: 2009-2010 EC
Budget
Budget code Budget (Birr)
2009 2010
6114 45000 30000
6217 10000 13000
6218 45000 6000
6219 20000 4500
6221 9000 12000
6223 19000 17000
6231 19800 16900
6241 10000 15000
6244 36000 0
6256 12000 13600
Total 227809 143000
Grand total 370809

Tentative work plan 2008 EC
No. Activity Unit Quantity/ Months Remarks
No. of Jul Aug. Sept. Oct. Nov. Dec. Jan. Feb. Mar. Apr. May Jun.
samples/
parameters
1 Comparison of periphyton and feed-based fish No. of experiments

farming in growth performance of O.niloticus in x x
2*4= 8
ponds (Kibru, Abelneh, Fikadu-T, Fikadu-HM)
 Pond preparation, substrate collection no. of ponds/ no.of 9/3 x
substrate plants
 Periphyton production optimization experiments no.of days 60 x x
 Fish growth performance and water qualityno.of days 240 x
dynamics experiment
 Water quality evaluation no. of measurements 20 x x x
 Laboratory analysis x x x x x x x x
Expected outputs:
Alternative nature-based fish feed technology for small-scale fish farming
References
APHA (1999) American Public Health Association: Standard methods for the examination of water and waste
water. 20.ed. Washington, D.C: APHA, 1999. 1325p.
Azim M. E., Verdegem M. C. J., Khatoon H., Wahab M. A., van Dam A. A. and Beveridge, M. C. M. (2002)
A comparison of fertilization, feeding and three periphyton substrates for increasing fish production in
freshwater pond aquaculture in Bangladesh. Aquaculture, 212:227-243.
Azim, M.E., *, M.M. Rahamanc, M.A. Wahabc, T. Asaedaa, D.C. Littled, M.C.J. Verdegemb
(2004b)/Periphyton-based pond polyculture system: a bioeconomic comparison of on-farm and on-
station trials. Aquaculture. 242 (2004) 381–396
Edwards, P., 1993. Environmental issues in integrated agriculture–aquaculture and wastewater fed fish culture
systems. In: Pullin, R.S.V., Rosenthal, H., Maclean, J.L. (Eds.), Environment and Aquaculture in
Developing Countries, ICLARM Conf. Proc., vol. 31, pp. 139–170.
El-Sayed, A.F.M (1998) Total replacement of fishmeal with animal protein sources in Nile tilapia
Oreochromis niloticus (L) feeds. Aqauculture. Res., 29:275-280.
Dugan I. (2001) The ecology of periphytic rotifers. Hydrobiologia 446/447: 139–148.
Degefu, F. (2011) Modelling periphyton based fish production. Journal of Agricultural Development. 2(1): 1-
17.
Degefu F., S. Mengistu, M. Schagel (2011) Influence of fish cage farming on water quality and plankton in
fish ponds: A case study in the Rift Valley and North Shoa reservoirs, Ethiopia. Aquaculture. 316:(1–
4) 129–135.
FAO. (2010a). State of World Fisheries and Aquaculture 2010. Food and Agricultural Organization of the
United Nations. Available at: http://www.fao.org/docrep/013/i1820e/i1820e00.htm
Hunt, B.P.(1952) Food relationship between Florida spotted gar and other organisms in the Tamiami Canal,
Dade Country, Florida. Trans. Amer. Fish. Soc., 82: 13-33.
Yared, T. (2010)Stocking based fishery enhancement programmes in Ethiopia. Ecohydrology &
Hydrobiology. 10( 2–4): 241-246.
Young, O. W.(1945) A limnological investigation of periphyton in Douglass lake, Michigan. Trans. Amer.
Micr. Soc., 64:1-20.

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Project IV: Development and evaluation of alternative feed source for

Background and justification

 To collect and culture Azolla species in laboratory and open field.

Materials and Methods

Phase one: Azolla sample collection, identification and harvesting activities

Use of a natural aquatic fern as

Dig small ponds for fern x x

Transfer of collected ferns into x x x x x x x x x x

Mass propagation and care of x x x x x x x x

Purchase of plastic trays for fern 01-15 01-15

Drying & Chemical analysis of x x x

Feed formulation for feed x x

Collection of male tilapia x

Feeding of tilapia with fern feeds x x x x x x

Budget by code for 2009 E.C.

Monitoring and evaluation matrix

Method of Tools for

 To adapt the culture and mass production techniques of earth worms.

 To experimentally test the acceptability of live worms and importance of dried

Materials and methods

 10% with worm, with tilapia carcass

 15% with worm, with tilapia carcass

Work plan for 2009

Use of earthworm as feed

Construction of shelter house x x x X

Proximate chemical analysis of x x X

Collection of catfish fingerling x X

Live feeding of catfish x X x x x x

Budget by code for 2009 E.C.

 Mass culturing (Vermicultre) of worms will be adopted.

 Use of worms as single feed will be experimentally tasted.

 Use of worms as fish feed ingredients will be evaluated.

Responsibilities (persons, centers)

National Fisheries & Aquatic Life Research Center (NFALRC), Sebeta

Method of Tools for

 Aguilar-Manjarrez, J. & Nath, S.S. (1998) A Strategic Reassessment of Fish Farming

Activity 3: Evaluate culturing techniques of the freshwater rotifer, Brachionus calyciflorus

Background and justification

Brachionus calyciflorus stock culture preparation

Application of rotifers to fish larviculture

Algal culture preparation X

Rotifer stock preparation X

Culturing Brachionus calyciflorus X

Running the experiment & larval feeding X

Data collection & data entry X

Budget codes 2009

• Batch culturing technique of rotifer adopted

• Guide line for larval fish live food feeding available

Center (s): NFALRC

Person (s): Adamneh Dagne, Kibru Teshome, Esayas Alemayehu

Output M&E Indicators Informatio Method of Tools for Method of

Background and Justification

 To adopt fish silage preparation technique

Growth and feed utilization parameters

a) Average weight gain (g/fish) (AWG):

Where: Wl = the initial weight (g)

W2 = the final weight (g)

b) Average daily gain (ADG)