Effects of four different doses of organic manures in the production of Ceriodaphnia cornuta

Bioresource Technology 97 (2006) 1036–1040

Effects of four different doses of organic manures in theproduction of Ceriodaphnia cornuta

Ashutosh Srivastava, Raja Mansingh Rathore, Rina Chakrabarti *

Aqua Research Lab, Department of Zoology, University of Delhi, Delhi 110 007, India

Received 28 January 2005; received in revised form 28 April 2005; accepted 29 April 2005

Available online 28 June 2005

Abstract

Mass culture of Ceriodaphnia cornuta was done by using a mixture of organic manures: cattle manure:poultry droppings:mustard

oil cake (1:1:1) at four different doses: 0.263 kg/m3 (first dose), 0.526 kg/m3 (second dose), 1.052 kg/m3 (third dose) and 2.104 kg/m3

(fourth dose). The peak of C. cornuta was found on 10th day of inoculum in first two doses and on 14th and 18th day in third and

fourth doses, respectively. Among these four doses, significantly (P < 0.01) higher numbers of organisms (1930/l) were found in the

fourth dose followed by third (1470/l), second (1017/l) and first (733/l) doses, respectively. The number of organisms decreased faster

in two lower doses than higher doses. pH ranged from 7.20 to 8.09, 7.46 to 8.01, 7.55 to 7.89 and 7.61 to 8.03 in first, second, third

and fourth doses, respectively. Dissolved oxygen showed inverse relationship with the dose of manures applied and direct relation-

ship with number of organisms. This study showed that 3.28–4.63 mg/l dissolved oxygen was optimum to obtain the bloom of C.

cornuta under the present manure schedule. Maximum number of organism was found when unionized ammonia and phosphate

levels ranged between 0.65–0.85 mg/l and 0.42–0.98 mg/l, respectively. The fourth dose of organic manure is optimum for the culture

of C. cornuta in outdoor condition and the bloom of the live food can be obtained within 18 days of inoculum.

� 2005 Elsevier Ltd. All rights reserved.

Keywords: Ammonia; Ceriodaphnia cornuta; Dissolved oxygen; Organic manures

1. Introduction

The Cladocera Ceriodaphnia cornuta (Class: Crusta-

cea, Order: Cladocera) popularly known as water flea,is a preferable food item of first feeding larvae of many

freshwater fishes due to their suitable size (258–588 lm)

for gape limited larvae. Several studies have documented

the size-selectivity of fish for their invertebrate food

(Brooks and Dodson, 1965; Dodson, 1974; Zaret,

1980; O�Brien, 1987). Cladocerans are easily digestible

due to the presence of digestive enzymes (Kumar

et al., 2005) and they have high energetic caloric value(Morris and Mischke, 1999). Gut content analysis of

0960-8524/$ - see front matter � 2005 Elsevier Ltd. All rights reserved.

doi:10.1016/j.biortech.2005.04.044

* Corresponding author. Tel.: +91 11 27666496.

E-mail address: [email protected] (R. Chakrabarti).

fourth day post hatch larvae of Indian major carp

showed the presence of this organism (Kumar, 2002)

in digestive system. Many studies (Sharma and Chak-

rabarti, 1998, 2000; Kumar et al., 2000; Rathore et al.,in press) evidenced the successful rearing of Indian

major carps and exotic carps, fed exogenously produced

mixed live zooplankton dominated by Ceriodaphnia spp.

during early ontogeny.

Most cladocerans remove particulate organic matter

from water by filtration. The size of particles that can

be cleared from the water is a function of the morphol-

ogy of the setae on the moving appendages (He et al.,2001). The ability of the cladocerans to ingest food of

a wider size range, and their higher filtering rates, give

them a better competitive edge over the rotifers (Allan,

1976; Lynch, 1980). Daphnia spp. with the help of spe-

cialized combs of setae on the thoracic appendages,

mailto:[email protected]

A. Srivastava et al. / Bioresource Technology 97 (2006) 1036–1040 1037

can utilize algae, bacteria, fungi, protozoa and organic

debris and even small food particles of 1–60 lm (Jorgen-

sen, 1966). Jana and Chakrabarti (1993a) also found the

presence of organic detritus in the gut of Moina micrura.

In the culture of larval fish of various species, manage-

ment of the zooplankton forage base is critical to suc-cessful transition of larvae to fingerling stage. The

information regarding the relative status of plankton

communities gives insight into water quality parameters

and the possible success of culture season (Morris and

Mischke, 1999). The phytoplankton and zooplankton

communities, which develop in rearing ponds, are influ-

enced by interactions between temperature, photo-

period, water quality, nutrient availability and fishpredation (Geiger, 1983). Liquid manures were highly

effective for culturing natural forage suitable for fish

growth (Schluter and Groeneweg, 1981). Animal man-

ures like cattle manure, poultry droppings and agricul-

tural by-products like mustard oil cake are wastes in

India. These can be used as potential organic manures

to increase the productivity of water body. Application

of organic manures influenced water quality. Jana andChakrabarti (1993a) found interactions between chemi-

cal oxygen demand, dissolved oxygen, phosphate levels

and primary productivity resulting in optimal conditions

were responsible for large variations in life history char-

acteristics of M. micrura. The present investigation aims

to study the effects of four different doses of organic

manures on the production of C. cornuta and linking

the changes in production over time with changes inwater quality parameters. Live food produced by using

optimum dose will be utilized for intensive larval rearing

as well as feeding of adult aquarium fish.

Fig. 1. Abundance of Ceriodaphnia cornuta in four culture systems

varying in doses of manure. Each point represents the mean of values

derived from three replicates.

2. Methods

Mass culture of C. cornuta was done by using amixture of organic manures: cattle manure:poultry

droppings:mustard oil cake (1:1:1) in 0.45 m3 outdoor

plastic tanks. Organic manures were used in four differ-

ent doses: 0.263 kg/m3 (first dose), 0.526 kg/m3 (second

dose), 1.052 kg/m3 (third dose) and 2.104 kg/m3 (fourth

dose). Three replicates were used for each treatment.

Organic manures were decomposed for 12 days before

the inoculation of zooplankton. This manure–waterwas mixed thoroughly daily manually. There was no

additional feeding of C. cornuta with phytoplankton.

and no aeration in the plankton culture tanks. On

13th day, C. cornuta were inoculated @ 20/l. Water

quality parameters such as temperature, pH (HANNA-

HI 8224), dissolved oxygen (Orion DO Meter-810), un-

ionized ammonia (Orion Ionalyser-720), nitrite, nitrate

and phosphate (APHA, 1995) were measured at weeklyinterval from each tank. Sub-samples of C. cornuta were

collected on every alternate day. The water containing

live food were thoroughly mixed before sampling. Five

liters of zooplankton rich water was collected from five

different sites of tank and was passed through plankton

net (mesh size: 75 lm). It was preserved in 4% formalin

and was concentrated to 10 ml. One milliliter of concen-

trated plankton sample was taken in Sedgwick raftercounter chamber and the number of C. cornuta was

counted under microscope and it was expressed as no./

l of water.

2.1. Statistical analysis

Data of plankton abundance and water quality

parameters were compared using one-way analysis ofvariance (ANOVA) and Duncan�s multiple range test

(Montgomery, 1984). Statistical significance was ac-

cepted at P < 0.05 levels.

3. Results and discussion

In the first dose (0.263 kg/m3), the peak of C. cornutawas found on 10th day of inoculum (733/l) and gradu-

ally the number of plankton declined. Peak appeared

on 10th day in the second dose (1017/l), whereas in the

third dose, the peak was found on 14th day of inoculum.

But the number of organism was 31–50% higher in

the third dose (1470/l) than first two doses. In the fourth

dose, the peak was found on 18th day of inoculum

(1930/l). Among these four doses, significantly (P <0.01) higher number of organism was found in the

fourth dose followed by the third, second and first doses

(Fig. 1). But the peak of plankton was found 4–8 days

earlier in first two doses than others. The number of

organism decreased faster in lower doses than higher

doses. The information about time required to obtain

the peaks in various doses used in the present study have

aquacultural importance. The fish farmer can manipu-late the dose of organic manures depending on their

requirement to feed the fish. The dynamic characteristics

1038 A. Srivastava et al. / Bioresource Technology 97 (2006) 1036–1040

of zooplankton populations have led the use particular

fertilization techniques and species-specific zooplankton

inoculations in culture ponds (Colura and Matlock,

1983; Geiger, 1983; Farquhar, 1984; Turner, 1984; Gei-

ger et al., 1985). In static batch culture,Moina mongolica

were fed with Nannochloropsis oculata and the density ofM. mongolica reached 540–3500 ind/l during 28 days

culture period (He et al., 2001). The food supply could

be a limiting factor in this type of culture due to the high

filtering rate. It is difficult to maintain the density of this

species >1000/l by using algae as sole food supply. Har-

vesting is recommended when the animal density reaches

500–1000/l. In the present study, 733–1930 C. cornuta/l

was obtained using only organic manures. This wascommercially viable, especially in the present scenario

of organic farming.

In this experiment, organic manures were decom-

posed for initial 12 days before the inoculum of C. corn-

uta. In a study of life table responses of M. micrura and

Daphnia carinata, Jana and Chakrabarti (1993b) sug-

gested that the interval between 2 and 10 days after

manure application is a prerequisite for establishingsatisfactory environmental conditions for zooplankton

production. Water temperature ranged from 26.83 to

30.5 �C throughout the culture period. There was no dif-

ference in water temperature among the treatments. pH

ranged from 7.20 to 8.09, 7.46 to 8.01, 7.55 to 7.89 and

7.61 to 8.03 in first, second, third and fourth doses,

respectively. There was direct relationship between the

dose of manure applied and free carbon dioxide levelin the present study. Free carbon dioxide was signifi-

cantly (P < 0.01) higher in the fourth dose than other

three treatments throughout the study period (Fig. 2).

Dissolved oxygen showed inverse relationship with

the dose of manures applied in the culture systems. Dis-

solved oxygen ranged from 4.63 to 6.43, 4.08 to 5.94,

3.08 to 5.43 and 1.27 to 4.63 mg/l in first, second, third

0

10

20

30

40

50

60

70

10 24 41 45

Days after inoculum

Fre

e C

arbo

n di

oxid

e (m

g/l)

First Dose Second Dose Third Dose Fourth Dose

17

Fig. 2. Free carbon dioxide levels of water in four different treatments

of Ceriodaphnia cornuta during 45 days of culture period. Each point

represents the mean of values derived from three replicates.

and fourth doses, respectively. Direct pond fertilization

using excessive manure can lead to water quality deteri-

oration (Boyd, 1982), including severe depletion of dis-

solved oxygen and high chemical oxygen demand of

water. Similarly, in the culture tanks of C. cornuta, dis-

solved oxygen level was minimum and maximum in firstand fourth doses, respectively. Dissolved oxygen was

minimum at the beginning of the study and maximum

at the end of the experiment in all these treatments

(Fig. 3). This data supports the findings of earlier

researchers. Jana and Chakrabarti (1993b) found that

dissolved oxygen level depleted to about half of its con-

trol value on second day of manure application, but im-

proved gradually as the days after manure applicationincreased, showing a nearly 2-fold rise on 11th day after

application. Morris and Mischke (1999) suggest that or-

ganic fertilizers may cause dissolved oxygen problems

during initial decomposition. There was direct relation-

ship between dissolved oxygen and number of organism.

In the first and second doses the dissolved oxygen levels

were 4.63 and 4.08 mg/l on day 10 when the maximum

no. of organisms were found. In the third dose the dis-solved oxygen level was around 4.32 mg/l when the

bloom of C. cornuta was obtained. In the fourth dose,

dissolved oxygen was 1.27 mg/l in the first week and

the number of organisms were 180/l. Gradually, the

amount of dissolved oxygen increased and when it

reached 3.28 mg/l, the peak of zooplankton (1930/l)

was obtained. Therefore, the range of dissolved oxygen

level should be maintained between 3.28 and 4.63 mg/l.So, the production of C. cornuta was inhibited when

the concentration of dissolved oxygen was lower. As

dissolved oxygen level increased in the culture system

with the progress of culture period, number of organism

also increased. The decomposition process encountered

immediately after manure application is highly adverse

to aquatic life and it is likely that populations of zoo-

0

1

2

3

4

5

6

7

8

10 17 24 41 45

Days after inoculum


Dis

solv

ed o

xyge

n (m

g/l)

Fig. 3. Oxygen levels of water in four different treatments of

Ceriodaphnia cornuta during 45 days of culture period. Each point


0

2

4

6

8

10

12

10 17 24 41 45

Days after inoculum

Nitr

ite (

mg/

l)


Fig. 5. Nitrite levels of water in four different treatments of



A. Srivastava et al. / Bioresource Technology 97 (2006) 1036–1040 1039

plankton are largely altered due to environmental per-

turbations (Jana and Chakrabarti, 1993b). Here, in the

initial days of manuring, the decomposition of manures

required more dissolved oxygen, which resulted in the

depletion of dissolved oxygen in the culture system. Bio-

logical oxidation converts the organic matter in manureto carbon dioxide. This free carbon dioxide quickly

combines in water to form carbonic acid, a weak acid.

Therefore, at the initial days of plankton culture, pH

and dissolved oxygen values were low and free carbon

dioxide was higher. This is related with the doses of

manures. The manures decomposed with time and the

toxic effects gradually reduced. This led to the abun-

dance of C. cornuta.Ammonia and nitrite are the main nitrogen com-

pounds considered risk factors in the growth of crusta-

ceans (Mevel and Chamroux, 1981). When manures

decay, ammonia is released and its nitrification produces

acidity (Boyd and Tucker, 1998). In this present study,

unionized ammonia (NH3) level was significantly

(P < 0.01) higher in the fourth dose than others (Fig. 4).

In all these treatments, unionized ammonia was higherat the beginning of the study and gradually ammonia le-

vel decreased. In the third and fourth doses, ammonia

levels were 9.69 and 17.44 mg/l, respectively on 10th

day of culture. When ammonia levels reached 0.723

and 0.85 mg/l in these two treatments, the number of

zooplankton become 1470 and 1930/l in third and fourth

doses, respectively. Whereas in the first and second

doses the highest no of C. cornuta was found whenammonia levels were 0.69 and 0.65 mg/l, respectively.

The optimum level of ammonia ranged between 0.65

and 0.85 mg/l in the present culture conditions. So this

is the optimum range of NH3 for production of C. corn-

uta. Schluter and Groeneweg (1985) found that in rotifer

Brachionus rubens, the reproduction rate decreased at

ammonia levels of 3–5 mg/l. The rotifer died within

0

2

4

6

8

10

12

14

16

18

20

10 17 24 41 45

Days after inoculum

Am

mon

ia (

mg/

l)


Fig. 4. Ammonia levels of water in four different treatments of



2 days at ammonia concentrations above 5 mg/l. Likeammonia, nitrite level was minimum in the first dose

and this was higher in the fourth dose (Fig. 5). Nitrate

level was minimum at the beginning of the study regard-

less of culture systems.

In the first, second, third and fourth doses, bloom of

live food organism were obtained when the phosphate

levels were 0.74, 0.98, 0.42 and 0.55 mg/l, respectively.

In first and second doses, phosphate level was signifi-cantly (P < 0.05) higher on 17th day of inoculum,

whereas, in third and fourth doses, significantly higher

amount of phosphate was obtained on 10th day of inoc-

ulum (Fig. 6). Phosphate level was minimum on 44th

day of inoculum in all treatments, except the fourth dose

whereas, minimum level was obtained on 24th day of

inoculum.

In conclusion, the fourth dose of organic manure wasoptimum for the culture of C. cornuta in outdoor condi-

tion and the bloom of the live food could be obtained

within 18 days of inoculum. This information would

be helpful for the farmer in managing the farm.

0

1

2

3

4

10 17 24 41 45

Days after inoculum

Pho

spha

te (

mg/

l)


Fig. 6. Phosphate levels of water in four different treatments of



1040 A. Srivastava et al. / Bioresource Technology 97 (2006) 1036–1040

Acknowledgements

Indian Council of Agricultural Research (I.C.A.R.),

New Delhi supported this research.

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Documents

Effects of four different doses of organic manures in the production of Ceriodaphnia cornuta