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1Raw waste water (RWW) 2 Treated waste water (TWW)
As shown in table 2, the incorporation of bacterial cells in the rearing tanks increased significantly (P<0.05) the mean weight increments for Common Carp larvae in T1 (rearing in treated waste water) in comparison with T2 (rearing in raw waste water).
The fishes of bioremediated treatment also recorded higher mean individual length increments in comparison with T2 and T3 (rearing in tap water).
The reared fish larvae in tap water showed a significantly (P<0.05) greater condition factor (1.96) and Specific growth rate (SGR), but the lowest of condition factor was in bioremediated treatment (T1).
Table2. The values of growth parameters of Common Carp larvae in different treatments
1 Food conversion ratio (FCR) = food intake (g) / living weight gain (g)
2Specific growth rate (SGR) = 100x [ln final weight of fish-ln initial weight of fish)]/days of feeding. 3Condition factor (CF) = 100x [(g final weight of fish)/ (total length of fish- cm) 3].
4Averagr daily growth (ADG) =100x [(final weight of fish- initial weight of fish) / (initial weight of fish).days of feeding].
Administration of the Bacillus sp. via direct inoculation to rearing tanks resulted significant higher growth and feeding performance in comparison with reared Carp larvae in untreated waste water. Microorganisms play important roles in the natural aquatic environment. They contribute to the geological cycle of elements and transformation of natural chemicals. Microorganisms are extremely diverse and are capable of utilizing the contaminant as an energy and carbon source to survive in inhospitable environments (Watanabe, 2002). Moreover, Bacilli use ammonium ion as the nitrogen source for its growth under aerobic conditions and, hence, are actively involved in nitrification in aquatic systems (Koops and Moller 1992). In this study the Total ammonium
Nitrogen, significantly decreased in bacterial treatment. The content of total ammonium Nitrogen decreased from 2.78 to 1.01 mg/l. The results
of this study clearly demonstrate that the bioremediation of waste water of carp cultivation system had good potential in exploitation and re-use of this sewage for cultivation of Common carp larvae in tank culture. The inoculation of belend of Bacillus licheniformisi, B. subtilis, B. polymyxa, B. laterosporus and B. circulans to the culturing tank increased the growth rate of Common Carp larvae but the best result were obtained in tape water (clean water). This is in agreement with the findings by Jafaryan et al., (2007) who also reported higher growth parameters for Persian Sturgeon (Acipenser persicus) larvae. In contrast of with our results, Jafaryan,et al., (2011) indicated that the addition of probiotic bacilli to rearing tanks water had negative effects on the growth parameters of Big head larvae when they were fed on different Artemia nauplii. This suggests that the addition of probiotics optimized the feed consumption and promoted growth parameters (Lara-Flores et al., 2003). In contradiction with our results Boyd et al., (1984) reported that the adding commercial probiotic bacteria didn't have any significant effect on growth parameters of channel catfish. Addition of bacteria bioencapsulated in Artemia metanauplii to a rearing system for halibut larvae (Hippoglossus hippoglossus L) didin't increased the growth parameters of this fish (Makridis, et al., 2001).
In agreement with these results Ziaei-Nejad et al., (2006) reported that when the probiotic bacilli were added to rearing tanks at the concentration of 7.3x106 CFU/ml, increased the growth parameters of Indian white shrimp (Fenneropenaeus indicus) in comparison with control.
However the present results indicated that the inoculation of probiotic bacilli to effluent or waste water of fish cultivation system, in processing of microbial bioremediation can use as a potential for reuse in rearing tanks of Common Carp (Cyprinus carpio) larvae.
Boyd, C.E., Hollerman, W. D., Plumb, J. A., Saeed, M. (1984) Effect of treatment with a commercial bacterial suspension on water quality in channel catfish (Ictalurus punctatus) ponds. Prog fish-Culture. 46: 36-40.
Gomez-Gil, B., Herrera- Vega, M. A., Aberu- Grobis, F.A, Roque, A. 1998. Bioencapsulation of two different vibrio species in nauplii of the Brine shrimp
(Artemia fransiscana). Appl. Envirn. Microbiol. 64: 2318- 2322.
Jafaryan, H., G. A. Takami, A. Kamali, H. Soltani and M. Habibirezaei. 2007. The use of probiotic bacillus bioencapsulated with Artemia urmiana nauplii for the growth and survival in Acipenser persicus larva. J. Gorgan Univ. Agricult. Sci.
and Natu. Res. (Iran). 14: 87-97.
Jafaryan, H., Soltani, M., Noferesti, H. and Ebrahimi, P. 2011. Effect of adding probiotics into the rearing tanks of grass carp (Ctenopharyngodon idell ) for the exploitation of Artemia urmiana, Artemia fransiscana and Artemia parthenogenetica Nauplii. International Journal of Veterinary Research. 3: 125128.
Koops, HP. and Moller, UC. 1992. The lithotrophic ammonia-oxidizing bacteria. In: Balows A, Truper HG, Dworkin M, Harder W, Schleifer K-H (eds) The prokaryotes, vol III. Springer, Berlin, Germany, pp 2625-2637.
Lara-Flores, M., Olvera-Novoa, Miguel A., Guzman-Mendez, Beatriz E., Lopez-Madrid, W. (2003) Use of the bacteria Streptococcus faecium and Lactobacillus acidophilus, and the yeast Saccharomyces cerevisiae as growth promoters in Nile tilapia (Oreochromis niloticus). Aquaculture. 216: 193-201.
Makridis, P., Bergh,Q., Skjermoj, J. and Vadstein, O. 2001. Addition of bacteria bioencapsulated in Artemia metanauplii to a rearing system for halibut larvae. Aquacult. Inter. 9: 225- 235 .
Nogami K. Maeda, M. (1992) Bacteria as biocontrol agents for rearing larvae of the crab Portunus trituberculatus. Canadian J. of Fish. and Aqua.Sci. 49: 23732376.
Olafsen J.A. Hansen, G.H. (1992) Intact antigen uptake in intestinal epithelial cells of marine fish larvae. J. of Fish Biolog. 40: 141-156.
Singh, I. S. B., N. S. Jayaprakash, P. Somnath. 2001.Antagonistic bacteria as gut probiotics. IP 24. p. 55-59. National Workshop on Aquaculture Medicine, January18- 20, 2001. Abstracts, CFDDM, SES, CUSAT, India.
Watanabe K (2002). Linking genetics, physiology and ecology: an interdisciplinary approach for advancing bioremediation. J. Biosci. Bioeng. 94 (6): 557-562.
Ziaei-Nejad, S., Habibi Rezaei, M., Azari Takami, G., Lovett, D.L., Mirvaghefi , A.R. Shakouri, M. (2006) The effect of Bacillus spp. Bacteria used as probiotics on digestive enzyme activity, survival and growth in the Indian white shrimp Fenneropenaeus indicus. Aquaculture. 252: 516-524.
Improvement of the Induced Spawning of Bunnei (Barbus sharpeyi, Gunther, 1874) in Iraqi Hatcheries
Sajed S. Al-Noor2, Mustafa A. Al Mukhtar1, Nadir A. Salman2, Malik H. Ali1
1 Marine Science Center, Basrah University, Basrah-Iraq
2 College of Agriculture, Basrah University.
Bunnei (Barbus sharpeyi) is one of the important local fishes in Tigris-Euphrates Basin, and the most important species in the marshes of this basin. Recently its population has sharply been declined in the marshes of southern Iraq due to the drying crime. The enhancement of this population needs the use of induced spawning to produce a good quantity of fingerlings to be released in the marshes. This article will discuss in details the technical steps for the induced spawning of this species, conducted in the fish hatchery of Marine Science Center (Basrah University, Iraq). The brooders were captured from AL-Huwaizha marsh and reared and fed in earthen ponds for three months with artificial ration. They were proved to be suitable for induced spawning. The suitable time for artificial breeding extends from the first week of March till the last week of April. Females received two doses of pituitary gland extract, a preparatory dose of 0.5 mg/ kg and a second dose of 3.5 mg/kg, while the dosage for the male was 3-3.5mg/kg injected at the time of the females second dose. An artificial hormonal preparation (OVAPRIME) was also used for the induced spawning with dosage of 0.5 ml/kg, giving better results for Bunnei induced spawning. An average weight of 1800g of fertilized eggs was obtained from the injected female. It was found that the best brooders weight for female was 750-1000g and for male 500-900g. The suitable hatching water temperature ranged between 22-24 C° with water flow rate of 600 mil/min. Both dry method of fertilization and use of fertilization solution were practiced. Tannic acid was used for the elimination of egg stickiness. The larval rearing and feeding step practiced in this experiment proved to be suitable both in using the live food and artificial diet.
The Iraqi marshes have been recognized to be the largest wetland in the Middle East and it has regional and global significance both for biodiversity and for human culture. These area cover 20000 square kilometer of open water, includes both permanent and seasonal marshes. The marshes provide important spawning, rearing and growing habitat for a wide range of economically- important fish species, such as Barbus sharpeyi, B. xauthepterus and B. grypus.
Bunnei (Barbus sharpeyi) is the most important species. Unlike other species, Barbus sharpeyi spawn only in shallow open water which is less than 75cm depth of the marshes (Jasim1988). Currently Bunnei fish stocks had changed dramatically in the recent decades due to the negative effect of the marsh drainage program (UNEP, 2001). The present marsh fisheries are dominated by the uneconomic species such as black and grey mullet and two introduced Carp species. After the restoration of the marshes, emphasis is laid upon restoration and enhancement of fish stock for the important local fishes such as Bunnei.
Some efforts have been done for culture of Bunnei in Iraq( Ali et al. 1885,1986; Farga and Chabak 1988; Al Nasih 1992). In Iran (Yazdi pour et al. 1994; Moazedi 2000) work on the propagation of this species. The main purpose of these efforts was the enhancement of Bunnei stock in the inland waters. All these efforts were succeeded to obtain good numbers of fingerlings.
Materials and Methods
Brooders for the hatchery were collected from AL-Huwaizah marsh which is considered as the most suitable site for Bunnei brooders (Fig.1) after studying fish catch composition of many sites. Hour AL-Dessam, a part of AL-Huwaizah marsh which was refolded recently, represents the exact location for collection. Water depth ranged between 0.5-2 m, salinity 2-3 ppt and pH 7.5-8.1. There are different types of aquatic plants such as Phragmites, Typha, Ceratophyllum, Vallisneris, Potamogeton, and Scripus with varying densities. Three stations were selected using
gill nets with 75X 75mm mesh size by local fishermen for brooder catch. They were kept in net cages with dimension of 1.5X1.5X2m, constructed from steel frame covered by net with mesh size 1.5X1.5 cm. Before transport, the morphometric characteristics of the mature Bunnei and sex dimorphisms of brooders were checked. Transportation was done by truck supplied with 1 ton tank aerated with oxygen bottles. In each trip 15-25 brooders were transported. The distance between the collection site and the hatchery approached 150 km. Transportation was conducted during the early morning.
A total of 693 fish were collected during the trial period. Brooders were then released in two earthen ponds constructed near the Marine Science Center (MSC) hatchery, with dimensions 45x 30x 1.7 m. Artificial diet was provided for the brooders twice a day, With the level of 5% of total biomass.
The composition of the diet was as follows:
Soya beans ................................. 25%
Minerals & Vitamins.................2%
Health condition of the brooders and water quality of the ponds were monitored daily. Fish maturity, fecundity and sex ratio were also checked during the rearing period of the brooders, along with condition factor and feeding status. Sex-separated brooders were kept inside the hatchery in (1.5 x 1.75 x 1 m) tanks supplied for 24 hr with oxygen from Oxygen generator.
Fig (1) The collection site (Alhuwaizah Marsh)
Results and Discussion
Collection period of brooders extended from 27/12/2004 to 30/4/2005. A total number of 693 fish were collected with weight ranging between 500-1500 kg and the length between 34-60 cm. Transportation trials showed that Bunnei brooders could be transported in a density ranging from 15 to 25 fish/ton of water. Low mortality rate of only 0.93% was recorded during transporting the brooders from fishing site to the hatchery. This confirms the possibility of using oxygenated plastic tank under suitable thermal conditions (18-24°C) for successful transport of brooders. However, upon keeping brooders in rearing tanks, saprolegniases (infection with Saprolegina fungus) caused nearly 5.19% mortality, especially among injured fish. Most of the injuries occurred as a result using gill nets for fishing, which is the only available fishing gear in this area. It was found that the smallest fishes were more tolerable to handling and transportation than bigger ones.
Monthly length frequency distribution of the collected Bunnei brooder
iran-Larvi, ll-l2 December 2Ol2-Karaj
in Al-Huwaizah Marsh during the period from January to April is shown in (Fig2). In January fish of 40-50 cm length group were dominated the length frequency of Bunnei brooders. The results showed also that, the large individual appeared at the beginning of the spawning season while small -sized fish appeared later on. This might be due to the fact that large individual mature early than the smaller one.
Fig. (2): Monthly length frequency distribution Alhuwaizah Marsh during January-April 2005
of Bunnei brooders collected from
Many criteria were used to select the suitable brooders for artificial breeding such as weight, length, shape of the cloacae, body depth, belly, good bright color and lack of external disease and parasite. Body shape of males differs from that of females in having more elongated body shape, smaller belly and less body depth, with narrowest cloacae.
The prepared artificial food ration was consumed successfully by Bunnei brooders in the rearing ponds, this has resulted in good condition
and survival of brooder for induced spawning activities. According to the data of Table (1) it seems that the ecological parameter of the rearing ponds were suitable for Bunnei brooders, as they resembled the natural values in the spawning area which were recorded by Jasim (1988). A remarkable behavior was noticed among, brooders in the rearing ponds. They swim mostly near the bottom during January and February and near the surface during March and April when water temperature increased (Table 1), especially at mid-day, and moving in schools. This behavior assists the brooders to be subjected to the required temperature for gonad maturation.
According to the previous experience with Bunnei species in Iraq it appeared difficult to spawn Bunnei artificially after being kept in rearing ponds (Ali et.al.,1985). In this trial, however, it was possible to get sex products from brooders reared in ponds for more than 2 months, due to maintaining suitable rearing conditions and efficient feeding. The hatchery was supplied with those brooders from mid February till 30 April at batches of 8-16 fish for both sex.
Table (1) Water quality of brooders rearing ponds during February-March 2005 (weekly average)
Successful breeding was achieved in 150 brooder from both sexes.
The brooders were injected either with Pituitary gland extract (P.G extract) or Ovaprime hormonal preparation. The Ovaprime was used for the first time for Bunnei, with a dosage of 0.5 ml/kg for both sexes in one injection. It has been found that eggs can be obtained after 12-14 hrs of injection, and the smaller size brooder showed more positive response for hormonal administration.. Injecting Ovaprime hormones caused 5.05% mortality among fish with more than 1kg weight. It appeared that the usage of Ovaprime is more easier than PG extract, because it decreased the handling process of the brooders by deceasing the induced spawning steps, as well as the time required for each spawning batch. P.G extract, on the other hand, was injected in two dosages. The first (preparatory dosage) was 0.3-0.5 mg/kg for females only. After 12 hr the second injection (final dosage) was given to both sexes which was 3-3.5 mg/kg, depending on water temperature. The behavior of the injected brooders was monitored continuously. After 12-13 hrs from the second injection the stripping process was conducted to get the sexual products. The average weight of the ova obtained was 1800 gr. (Table 2).
It was found that the most suitable period for getting sex products extended from the first week of March till the last week of April. Brooders with weight ranged 750 - 1000 kg for female and 500-900 kg for males are the most suitable for positive response to hormonal injection.
Dry fertilization method is more suitable for Bunnei, which mean the mixing of the sex products without water. Mixing was done by aid of feather for few seconds, and then the fertilization solution added, just covering the eggs. The fertilization solution had the same composition of the washing solution.
Table (2) Induced spawning results for Bunnei (Barbus sharpeyi) in MSC hatchery (2005)
NO. OF FEMALES
NO. OF MALES
The fertilized ova were washed with washing solution (160 NaCl + 120 Urea + 40 litter water from brooders tanks) for 10 minute with this solution followed by washing with water for 10 minute. This sequence was repeated three times. Then the fertilized ova were washed with tannin solution (10 gm/10 litter water) for 30 seconds followed by washing with water for 30 seconds. The fertilized ova were incubated in Zougar Jars (10 L) with an incubation density of 50-100 g for each Jar. Water flow rate was 600 ml/min, just enough for ova circulation. The incubation period extended from 72-96 hr at water temperature of 22-24°c . Dissolved oxygen concentration in the hatching jars approaching saturation at 10-12 ppm.
Hatched larva were transport to incubation tanks (0.75 m3) till complete absorption of yolk sac which extended for 24-36 hr. At post-larval stage, they were transferred to indoor rearing tanks and fed with live food (rotifers nupualii). The total number of larva produced was 600,000 with 40-60% survival rate. For fry production, indoor open water system has been constructed for rearing post larvae which contains
20 tanks (260-300 l. each) rearing density was 100 larvae/1.
Live food was used for feeding the post larvae, which includes rotifers (Brachionus sp.) and Artemia (Artemia salina) napulaii. In the first week the larval feeding was restricted to rotifers only. From the second week and on Artemia napualii were added to the feeding regime, with small amount of artificial supplementary food during the 4th week supplied twice daily. The amount depends on number of post larvae in the tanks. during the rearing period the growth rate of the post was monitored exhibiting promising weight increment due to well feeding and good water quality, which can be summarized as follows:
Water temperature (24 -26 C), pH (7.3 -7.9), Dissolved oxygen (9.3 -10.1 ppm), Salinity (1.2 - 1.6 ppt), Nitrate (0.25 - 0.5 ppm), Ammonia < 0.1 ppm.
Table (3) showed the length and weight increments of post larvae and fry till the last week of June. During July 2005 about 300,000 Bunnei fingerlings with good health and condition were released in selected station of Al Hammar marsh.
Table (3) Weight and length increment of Bunnei post larvae and fry in indoor rearing system during the period April-May 2005
LENGTH INCREM. (mm)