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Effect of Wolffia sp. as Supplemental Feed on Growth Performance and Body Composition of Indian Major Carp, Labeo rohita (Hamilton, 1822) Fingerlings

The effect of Wolffia sp. as supplemental feedon growth performance and the body composition of Indian major carp, Labeo rohita fingerlingswere examined. The experiments were carried out in the laboratory in triplicates. The commercial feed of tokyu was used as feed for all experimental fish. In addition to tokyu, 10 gram wet weight of Wolffia sp. was directly introduced as supplemental feed for all experimental fish except control. Good survival rate was observed in control (95%) and experiments (97%). At the end of experiments, the growth of all Wolffia sp. fed L. rohita showed significantly higher (p<0.05) than control fish. More significantly differentin weight gain, protein efficiency ratio, food conversion ratio, specific growth rate, means daily weight gain and condition factor were also found in Wolffia sp. fed L. rohita than that of control. The protein content of Wolffia sp. fed L. rohita was better than that of control fish. All experimental fish gave positively length weight relationships and L. rohita consumed Wolffia sp. throughout the experimentation was observed. The present study can be concluded that the use of Wolffia sp. improve the nutritional quality of L. rohita fingerlings.


Introduction
Fish feed are very important and of interest in the economic production of fish by farmers in the world. Each of the fish production countries has produced a variety of local traditional commercial fish diets. If commercial fish diets alone are used in the culture fish species. Though, feeding management plays a critical role in the success of fish culture (Murugesan et al., 2010).
Most commercial fish feed ingredients are costly so a lot of research has been undertaken to find out suitable and cheaper ingredients in fish feed. Fish farmers need plenty and cheaper fish feed than local traditional commercial fish diets. The advantages of live food are superior because of its nutritional value which promotes normal growth and good health. To obtain natural live food for cultured of many herbivorous fish species should be investigated. A number of plants were continuously investigated for their potential in supplementing or replacing fish meal (Amisah et al., 2009).
Aquatic weeds and algae of Lemma, water hyacinth, Hydrilla, Spirogyra, Wolffia, Ipomoea, Azolla and Elodia etc. are abundantly found in Mandalay Environs. It should be considered of these aquatic weeds and algae to use for the supplemental feed of some fish species. The duckweed (Family Lemnaceae), are mainly the rootless Wolffia arrhiza, which are able to rapid vegetative reproduction and adaptable to unfavorable  (Murugesan, Sivasubramanian and Altaff, 2010). Therefore Wolffia sp. was chosen and used as supplemental feed in this experiment because live feeds play a vital role in the enhancement of fish growth. The research on Wolffia arrhiza used as food and feed additives were used for supplemental feed replacing soybean meal in the diet of Nile Tilapia, Oreochromis niloticus (Chareontesprasit and Jiwyam, 2001). Labeo rohita is also chosen in this experiment because it is one of the economic cultured fish species in Myanmar fish farms.
Thus, the study on the effect of L. rohita fingerlings and Wolffia sp. as supplemental live food was investigated. The production of fish was often been significantly influenced by the quality and quantity of phytoplankton in fish ponds and plants of the Lemnaceae, called duckweeds and water meals has high protein content from 15 to 45%, making it an excellent material for animal, poultry, and fish feed (Padmavathi and Veeraiah, 2009). This research was carried out to investigate the growth response of L. rohita which were fed with the feed using Wolffia sp. as supplemental feed under laboratory.
Only dietary protein concentration in live food may show significant influence on fish growth and fish performance. The growth of Wolffia sp. can be a good resource of proteins and utilized for the production of value-added products especially supplement fish feed.
The objectives of experiment were to study on the growth of L. rohita fingerlings and to determine the body composition of L. rohita fingerlings due to the effect of Wolffia sp. as supplemental food.

Study Area and Period
The study was conducted at the laboratory of Zoology Department, University of Mandalay at Mandalay region. All experiments were carried out from June to November 2011.

Collection of fish and Wolffia sp.
The Labeo rohita fingerlings were collected from Thetyetkone Fish Culture Farm, Mandalay. They were brought alive immediately to laboratory in oxygenated plastic bags and taken immediately to the laboratory.
Fresh Wolffia sp. were collected from fish ponds at east of the Bawdikone Banyan Pagoda campus (Fig. 1), which were put into the sterile-bottles and taken immediately to the laboratory. On the day of sampling, taxonomic identification was  (1968) and Desikachary (1959). They were recorded as photomicrographs by using DP 12 Olympus digital camera.
Then the identified species were placed in glass aquaria (30 cm x 15 cm x 15 cm) containing 5 L tap water and they were kept under the sunlight to culture as supplement

Experimental design and diets
The experiments were carried out in rectangular six glass aquaria (60 cm x 30 cm x 30cm) each containing 40 L tap water and 15 L. rohita fingerlings were stocked.
Adequate aeration used by air pump and the level of water were maintained throughout the experimental period. All aquaria water was changed once a week to keep the water in good quality. There were triplicates in each treatment.
At the beginning of experiment, fish from all aquaria were fed with the diet tokyu (10% of initial total body weight) twice a day, during 9-10 am and during 1-2 pm ( Fig. 2 A). In addition, 10 ml (wet weight) of Wolffia sp.(smallest flowering plant) was introduced directly as supplemental food for all experimental fish except control during 4-5 hours (pm) (Fig. 2 B and C).

Growth Parameter
Fish mortality and the parameters of water quality such as temperature,

Statistical analyses
Data were analyzed by one way ANOVA (SPSS version 11.5) and was used to compare difference among group means.

Results
Growth performance and nutrient digestibility of Labeo rohita fingerlings fed with the experimental diets were shown in  At the end of experiments, faeces were observed to contain the incompletely undigested parts of Wolffia sp. especially cell wall and detritus (Fig. 2 D and E). The growth of L. rohita fingerlings in the Wolffia sp. supplemented food showed significantly higher (p<0.05) than controls. Comparative growth of all experimental fish was shown in Fig. 3 A. The length weight relationship was observed R 2 = 0.9228 in control fish and Wolffia fed fish was R 2 = 0.724 at the end of experiment (Fig. 3 B and C).
During the experiments, water temperature was observed within 20-22°C in the morning and 25-28°C in the evening, dissolved oxygen (DO) was 3 mg/L-5.8 mg/L and pH was within 7-8.5.

Discussion
At the end of experiments good survival rate (SR%) and low mortality (M%) were observed in all experimental fish of Labeo rohita fingerlings. The final means length (FML) and final means weight (FMW) of 14.49 ± 1.47 cm and 26.48 ± 6.48 g was recordedin Wolffia sp. fed L. rohita fingerlings and 11.56 ± 2.62 cm and 9.75 ± 2.89 g in control fish. The FML and FMW of Wolffia fed L. rohita fingerlings were significantly (p<0.05) higher than control fish.
According to the results more weight gain (WG) of 9.08 ± 7.4, specific growth rate (SGR) of 0.79 ± 0.4 and daily weight gain (DWG) of 8.34 ± 3.33 were found in L. rohita fingerlings fed with the diets Wolffia than in control fish WG of 4.85 ± 2.66, SGR of 0.69 ± 0.15 and DWG of 5.34 ± 0.95. Agreed with Chareontesprasit and Jiwam (2001), reported that some aquatic plants have been commonly used as supplementary feeds, e.g. high protein Azolla, Lemna whilst Wolffia, a floating aquatic plant with high protein contents could possibly do the same.
In Wolffia fed L. rohita fingerlings were found PER of 17.86 ± 13.63andFCR of tokyu (2.26 ± 2.65) and Wolffia sp. (0.72 ± 0.85). In control fish was also found PER of 1.43 ± 0.73 and FCR of tokyu was 0.44 ± 0.19. This result gave the well PER and lowest FCR were found in control fish and in diet Wolffia sp. fed L. rohita fingerlings. Therefore it may be assumed that experimental fish can be easily consumed of the Wolffia sp. as tokyu. Wolffiaarrhiza is favorite food of fish, therefore it is beneficial to pisciculture especially when they are young (Parmar and Patel, 2010). The body compositions of Wolffia sp. fed L. rohitafingerlings gave crude protein of 16.9% (wet) and 60.2% (dry), crude fat of 3.5% (wet) and 6.5% (dry), total ash of 4.5% (wet) and 12.5% (dry), moisture of 70.4% (wet) and 3.5% (dry). In the tissue of control fish also gave crude protein of 11.59% (wet) and 41.29% (dry), crude fat of 13.0% (wet) and 10.14% (dry), total ash of 15.43% (wet) and 23.86% (dry), moisture of 80.49% (wet) and 4.0% (dry).
According to the results, body composition of L. rohita fingerlings was recorded more crude protein, lower moisture, crude fat and total ash were found in Wolffia fed fish than that of control fish. The experimental diet of Wolffia sp. contained crude protein content and moisture of 3.06% and 95.87% in wet and 17.94% and 11.68% in dry.
The condition factors K value of 1.07±0.47 was found in Wolffia fed L. rohita fingerlings and that was 2.22±2.54 in control fish. The K value was not significantly difference between Wolffia sp. supplemented food fedL. rohita fingerlings and control fish. In fisheries science, the condition factor is used to compare the "condition" "fitness" or wellbeing of fish (Anene, 2005). Length weight relationship of L. rohita fingerlings was positively correlated with R 2 values of 0.724 for diets Wolffia fed fish and control fish was 0.9225. During the experimental period, the values of water temperature, DO and pH were more or less similar in all aquaria.
The faeces of Wolffia sp. fed fish expect control and the gills of death L. rohita were also examined. Faeces were observed to contain the incompletely undigested parts of Wolffia sp. especially cell wall and detritus ( Fig. 3D and E). Dead fish were suffered from Dactylogyrus sp. infection on gills filaments because of these parasites were During the experiment, more growth was found as well as non-polluted the water quality in all aquaria. Therefore Wolffia sp. could be used not only supplement feed but also aquatic plant in fish culture ponds. In addition the aquatic plant of Wolffia sp. is occurred annually found in many ponds and Lakes. Fish, like other organisms, required food (energy) in order to grow, survive and reproduce, the food items (source of energy) in aquatic habitat are in the form of plankton, periphyton, nueston, benthose, nekton and plants are available throughout the year (Narejo et al., 2010).
Thus it could be considered as a supplement feeding material for fish as a direct feeding stuff or could possibly be substituted in place of commercial feed. Furthermore, the market price of commercial feed could be relatively high and so other sources of high protein contents from aquatic plants should be substituting. As such, supplement alternative feeds should be used to reduce the cost of feed in the aquaculture industry. Therefore the aquatic plant Wolffia sp. could be used as supplementary feed for L. rohita fingerlings because Wolffia sp. could provide this amount of protein without any cost and could be easily obtained from the fish ponds.

Conclusion
The result found that the supplemental feed of Wolffia sp. was suitable feed for L. rohita fingerlings. According to the present study, Wolffia sp. should be used in improving the nutritional quality of practical feed of fish because more growth and good body composition gave in L. rohita fingerlings. It was readily accepted that the Wolffia sp. were observed to consume feed throughout the experimentation. On the basis of this investigation, it was concluded that Wolffia sp. could be considered as a feeding material for fish either as a direct feeding stuff or a source of protein for any formulated rations and it could possibly be substituted in any other formulated or commercial fish feed.