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Nannochloropsis species are widly used as food in aquaculture (Maruyama et al. 1986; Volkman et al. 1993; Apt & Behrens 1999 ; Suda et al. 2002 ), as it contains highly nutritional compounds such as sterols and polyunsaturated fatty acids ( Mohammady et al. 2005 ).

The main microalgae used in the hatchery is Nannochloropsis, which is

used to culture rotifers. It is also introduced into the larval rearing tanks to provide a food source for rotifers, as well as to serve as a buffering medium for homogenous light intensity and water turbidity ( sih-yang, S. et al. 2005), and this species was used to feed rotifers that culthured in Bandar Imam Khomaini marine fishes research station also. This study have been carried out to determine the best salinity and culture media for better rearing and further cocentration of N. oculata.

Material and Methods

This experiment has been carried out in Bandar Imam Marine Fishes Research Station, Khuzestan. The experiment includes 12 treatments with 4 salinities ( 20, 25, 30 and 35 ppt ) levels and 3 kinds of culture media ( Giulard F/2 , CONWAY and mSATO ). Each treatment include 3 replicates of 0.5 liter Erlens. Distribiuted between the treatments by completely randomize desighn (CRD). Waters with diferent salinities made from filterd sea water and distilled water. The water has been disinfected with 30 ppm chlorine for 24 hours, then dechlorinated with 15 ppm sodium thiosulphate. Disinfected water added into 0.5 liter erlens and autoclaved at 120 °C and pressure of2. lb/inh2 for 20 mins. When containers were cooled, culture media were dded into them and sterilized again unader the UV for one hour. 30 ml of exponentialy grown algae were added to culture flasks and stored them at an ambient temperature maintained at 23±1 °C. Cultures were continuously agitated by bubbling with stelized air. Illumination was provided by fluorescent lamps with an light intensity of 1500 to 4000 lux under a 16 h/ 8h light/dark regime ( Nagwa, G. E. D. et al. 2005; Lavens, P. & Sorgeloos, P., 1996). The algae cultured for 6 days under the this conditions.

Growth survey

Algae growth was surveid after 6 days by a cell count using a haemocytometer ( Fuchs-Rosenthal grid, 0.1 mm deep ) under a light microscope . Growth data was subjected to standard two-way Analysis of Variance ( two-way ANOVA ) using SPSS13 software. All means were

compared using Duncan methodwith with confidence level of p<0.05.

Results

Results from two-way ANOVA were shown that there was a significant difference (p<0.05) between cell concentration means for different culture media used in this experiment. The maximum concentration has been measured for CONWAY culture media which significantly higher than other culture media ( table 1 ).

Table 1 : two-way ANOVA ruselt of cell concentration of Nannochloropsis oculata at different culture media

Culture media

Mean of cell concentration(106 cells/ml)

CNWAY

95.10 ± 13.94b

Giulard F/2

84.46 ± 8.85a

mSATO

85.13 ± 8.99a

Mean ± SD : treatments with different supperscrit letter have a significant difference ( P < 0.05 ).

Also a significant difference (p<0.05) has been observed between the means of cell concentration at different tested salinities and the best concentration obtained was from 20 ppt salinity. There was no significantly difference between 20, 25 and 30 salinities but all of them were significantly higher than 35 salinity ( table 2 ).

Table 2 : two-way ANOVA ruselt of cell concentration of Nannochloropsis oculata at different salinities

Salinity ( ppm )

Mean of cell concentration(106 cells/ml)

20

93.44 ± 12.51 b

25

91.28 ± 10.25 ab

30

86.72 ± 13.01ab

35

81.47 ± 7.82 a

Mean ± SD : treatments with different alphabet have a significant difference ( P < 0.05 ).

The best cell concentration of N. oculata algae was obtained from treatment 1 that it was include CONWAY culture media and 20 ppt salinity although it was not significantly higher than other salinities with same culture media and 20 and 25 ppt with Guilard F/2 culture media and 25 ppt with mSATO culture media but it was significantly (p<0.05) higher than others ( table 3 ).

Table 3 -Cell concentration comparisonof N. oculata algae at different salinity and culture media.

No. of treat.

Treatment

Cell concentration ( milcell/ml )

1

CON. *20

105.92 ± 12.98 b

2

CON. * 25

91.42 ± 15.63 ab

3

CON. * 30

96.33 ± 17.83 ab

4

CON. * 35

86.75 ± 6.00 ab

5

F/2. *20

91.92 ± 6.28 ab

6

F/2. * 25

87.42 ± 7.38 ab

7

F/2. * 30

84.58 ± 10.76 a

8

F/2. * 35

76.92 ± 5.16 a

9

SATO. *20

82.50 ± 1.39 a

10

SATO. * 25

95.00 ± 10.04 ab

11

SATO. * 30

82.25 ± 5.97 a

12

SATO. * 35

80.75 ± 10.40 a

Mean ± SD : treatments with different alphabet have a significant difference ( P < 0.05 ).

Discussion

Analysis of six day results of growing period of N. oculata algae in different media and salinities showed that the best growth of this algae observed in treatment 1 ( CONWAY culture media * 20 ppt salinity ) although it was no significant difference between its concentration and the concentrations measured for 25 and 30 ppt salinities ( P > 0.05 ). So it,s recommendable to adopt these salinities to grow this species. We could not found any study about this topic but others workers use Guilard s F/2 culture media for growing this species ( Okumufi et al. 2002 ; Xan, L. & Hai, D. X. 2004 ) but this study showed that CONWAY culture media significantly ( P < 0.05 ) better than other

media.

Photosynthesis of algae is inhibited by osmotic stress ( Richmond, A. 2004 ) and the salinity play a main role at this stress. Each algae species appropriates an optimum salinity renge for optimum growth. In all of the studies have been carried out on N. oculata species , researchers used the salinity about 25 ppt to grow it ( Xan, L. & Hai. D. X. 2004 ). Results of

this study showed no significantly difference ( P > 0.05 ) in cell concentration in the salinities in the range of 20 to 30 for this species the result which agree with the results obtained from other studies, thus it possible to conclude " salinities in the range of 20 to 30 ppt in CONWAY culture media are the optimum salinities to grow N. oculata algae " , also for many species of microalgae salinity in the range of 24 to 26 ppt is adopted as an optimum renge for growth ( Lavens, P. & Sorgeloos , P. 1996 ) which clearly correspond to the results odf this study.

Acknowledgements

We thank Dr. Amin Ranjbar , of Bandar Imam Khomeini Marine Fish Research Station , for his helpful suggestions and information and to all other workers of this station for their very helpful aids.

References

Lavens, P. & Sorgeloos, P. 1996 . Manual on the production and use of live food for aquaculture. FAO Fisheries Technical paper. Rome .

Maruyama, I. ; Nakamura, T. : Matsubayashi, T. : Ando, Y. & Bl Maeda, T, 1986 . Identification of the algae known as marine chlorlla as a member of the Eustigmatophyceae. Japanese Journal of Phycology. 34 : pp. 167 - 172.

Mohammady, N. G. E. D. ; Chen, Y. C. ; El-Mahdy, A. R. A. & Mohammad, R. F. 2005

. Physiological responses of the eustigmatophyceae Nannochloropsis salona to aqueous diesel fuel pollution. Oceanalogia, 47 ( 1 ) : pp. 75 - 92

Okumuf , U. ; Baficiner , N. & Ozkan, M. 2002 . The effects of phytoplankton concentration , size of mussel and water temperature of feed consumption and filtration rate of the Mediterranean mussel ( Mytilus galloprovinvialis Lmk ). Turk J. Zool. 26 : pp. 167 - 172.

Richmond, A. 2004. Handbook of microalgae culture: Biotechnology and applied phycology. Blackwell science Ltd. 566 pp.

Sim, S. Y. ; Rimmer M. A. ; Toledo, J. D. ; Sugama, K, ; Rumengan, I. ; Williams, K. C. & Phillips, M. J. 2005 . A guide to samall-scale marine finfish hatchary technology. Australian center for international agricultureal research. 25 pp.

Suda, S. ; Atsumi, M. & Miyashita, H. 2002. Taxonomic characterization of a marine

Nannochloropsis species, N. oceanica sp. Nov. ( Eustigmatophyceae ).

Phycologia, May. 2002.

Volkman, J. K.; Brown, M. R.; Dun Stan, G. A. & Jeffrey, S. W. 1993. The biochemical composition of marine microalgae from the class Eustigmatophyceae. Journal of phycology. 29 : pp. 69 - 78.

Xan, L. & Hai, D. X. 2004 . Effect of temperature , salinity and stoching density on development of mass culture of Isochrysis galbana and Nannochloropsis oculata for breeding. Increasing Aquaculture Productivity , p. 23 ( abstract ).

Department of Natural Resources, Isfahan University of Technology, Isfahan, 84156­83111, Iran.

Introduction

Anesthetics are used with increasing frequency in aquaculture, especially to reduce the stress and mechanical injury to aquatic animals during handling (2). The use of anesthetic agents is particularly common during routine tasks such as biometry (3), transportation, and broodstock management (4).

Anesthesia is a biological state with the partial or complete loss of sensation or loss of voluntary neuromotor control included by chemical or nonchemical means (1). Increased time of exposure or concentration to anesthetic, first sedate an animal, than cause progressive loss of mobility, equilibrium, finally block reflex actions (4). Choosing an anesthetic must be attributed to several characteristics including its efficiency, availability, cost effectiveness, ease of use and safety for the user including fish, humans and the environment (7).

It is generally accepted that anesthesia maybe a useful way to maintain fish stress-free during essential manipulations in aquaculture. However, these agents are also known to cause stress (3), thus stimulating blood indicators of stress (8). In fact, anesthesia generally involves a cessation of breathing which, in turn, reduces gas transfer leading to hypoxia (1). As a result, it has been reported that anesthesia may lead to stress response through changing blood parameters in fish (8).

Clove powder is a dark brown powder from flowers, stalks and leaves of the clove tree Eugenina caryophyllata. Active ingredient is eugenol which has been widely used for several purposes such as antioxidant, antibacterial as well as anesthetic (2). Clove powder has some of

characteristics considered for an ideal anesthetic agent. Eugenal is a phenolic compound which inhibits the prostaglandin H synthesis and result in analgestic effects of clove powder (7). Eugenal-based anesthesia is effective at low dosages, inexpensive, easily obtained and safe for both environment and user. Therefore it could be a promising anesthetic agent in aquaculture, thus it is essential to investigate its anesthetic optimum dosage and effects on physiological parameters such as haematological characteristics at least in commercial fish species such as common carp, Cyprinus carpio.

The aim of this study is to compare some hematological indices such as red blood cell, RBC, White blood cell, WBC, hematocrit, hemoglobin between two anesthetic dosages in common carp as one of the most important commercially produced fish in Iran.

Materials and method:

The experiment conducted on juvenile common carp which were obtained from Isfahan culture and breeding centre, transported to the experimental aquarium in department of natural resources, Isfahan University of technology. Ten common carp weighting (mean weight 50.46 g mean length 15.45 cm) were housed in experimental aquarium and acclimated to it for a one week. Throughout the acclimatization period and during experiment, environmental conditions were monitored and maintained within optimum range of variable. A total 10 fish divided into two groups. In group 1 and 2, fish were anesthetized respectively using 500 and 750 mg/L clove powder. There were 5 fish in each group. These two different concentrations of clove powder (500 and 750 ppm) were chosen according to our pervious pilot study (not published data).

In each experimental group, fish were individually anesthetized with mentioned dosages. The observation of stage 3 anesthesia was considered as complete sedation (7, 10). Blood samples were taken just after complete sedation. Then, the fish was placed in freshwater tank for recovery. Sample preserved in disodium salt of ethylene diamine tetra-acetic acid (EDTA) bottles for analysis. The count of erythrocytes and

leukocytes were enumerated in an improved neubaeur hemocytometer, using hayem and truck diluting fluids (10). The amount of hemoglobin was determined according to cyanomethemoglobin procedure (7, 10). Hematocrit was determined by the standard microhematocrit method and expressed in percentage. The hematological indices (MCV, MCH, and MCHC) were calculated according to Svobodova et al. 1991 (11). The results are expressed as mean values + S.D. The data from experiments were analyzed by one-way ANOVA by using SPSS V.17 followed by student's T-test to compare the results of two experimental groups. In all cases, differences were considered at p<0.05.

Results

Effects of clove powder on the some hematological parameters are shown in Table 1. The fish which anesthetized with 750 mg/L showed higher values of Hct 9.40+0.78 % and RBC 0.78+0.12 (*106 mm-3) than those of in 500 mg/L 8.16+0.98 % and 0.57+0.05 (*106 mm-3) respectively (Table. 1). The rest of the indices (WBC, Hb, MCV, MCH, MCHC and PLT) didn't show any differences between two experimental groups (p>0.05).

Table 1: The effects of exposure to two dosages of clove powder on hematological parameters on Cyprinus carpio.

^^^^^ Group Parameters

500

750

RBC (*106mm-3)

0.57+0.05 a

0.78+0.12 b

WBC (* 103mm-3)

200.44+9.35

180.62+24.13

Hb (g dL-1)

8.88+1.00

9.40+0.78

Hct (%)

8.16+0.98 a

11.70+2.49 b

MCV (fl)

140.86+3.70

146.86+8.33

MCH (pg cell-1)

156.74+24.78

122.72+27.39

MCHC (g dl-1)

1.09+0.19

0.83+0.23

PLT (* 103 |iL-1)

80.75+28.46

135.75+64.79

Values are mean +S.D; values followed by different superscript letters in each row are significantly different at p<0.05 (ANOVA). MCV-mean corpuscular volume, MCH-mean corpuscular hemoglobin, MCHC-mean corpuscular hemoglobin concentrations, WBC-number of leukocytes, RBC- number of erythrocytes, Hb- hemoglobin, Hct hematocrit, PLT-number of plackets.

Discussion

In this study, the significant increase (p<0.05) of red blood cells and hematocrit values after clove powder anesthesia was observed in 750 ppm experimental group in compare to the other group. Haemotological profiles of blood can provide important information about their internal environment (2). In fact, the blood parameters of teleost fish may be affected by acute or chronic stress. The increase may result from the release of immature red cells by the spleen and could be an immediate response to the acute stress mediated by catecholamines (6). Similar results were found in fresh water and marine fish species using other anesthetic agents (6). Hypoxia as a result of reduction in respiratory actions lead in physiological changes in the blood factors such as raising hematocrit and RBC to combat whit lowering in O2 in circulation for breathing and survival (7). Similar results were found in RBC and Hct after using 2-phenoxyethanol as an anesthetic agent on common carp (2). The results indicates that increasing number of RBC, is the major factor which affect Hct value in the fish under defined experimental condition.

WBC was measured to evaluate clove powder effect in two mentioned dosage on fish immune system. It was showed that there was no significant difference between two groups, so, the results indicates releasing of WBC did not changed because of different dosage of clove powder as an anesthetic agent.

Conclusion

The data presented in this study reveal that use of clove powder at anesthetic dosage (500 and 750 ppm) have different effects on at least

some blood parameters. Since the RBC counts and Hct values was significantly higher in fish exposed to 750 ppm of clove powder, the recommended dose will be 500 ppm without to elicit hypothalamus-pituitary-internal axis (HPI) in common carp.

References

Gholipour Kanani H., Mirzargar S. S., Soltani M., Ahmadi M., Abrishamifar A., Bahonar A., Yousefi P. 2011. Anesthetic effect of tricaine methanesulfonate, clove oil and electroanesthesia on lysozyme activity of Oncorhynchus mykiss. Iranian Journal of Fisheries Sciences, 10: 393-402.

Velisek J., Svobodova Z., Piackova V., Groch L., Nepejchalova L. 2005. Effects of clove oil anesthesia on common carp (Cyprinus carpio).Veterinary Medicine

Czech, 6: 269-275.

Weber R.A., P6rez-Maceira J.J., Peleteiro J.B., Garcia-Martin L., Aldegunde M. 2011. Effects of acute exposure to 2-phenoxyethanol, clove oil, MS-222, and metomidate on primary and secondary stress responses in Senegalese sole (Solea senegalensis). Aquaculture, 321: 108-112.

Imanpour M.R., Bagheri T., Hedayati S.A.A. 2010. The anesthetic effects Clove essence in Persian Sturgeon, Acipenser persicus. World Journal of Fish and Marine Sciences, 2: 29-36.

Ross L.G., Ross B. 1999. Anaesthetic and sedative techniques for aquatic animals, 2nd ed. Blackwell Science Ltd., Oxford. 159 pp.

Weber R A., Peleteiro J.B., Garcia Martin L.O., Aldegunde M. 2009. The efficacy of 2-phenoxyethanol, metomidate, clove oil and MS-222 as anaesthetic agents in the Senegalese sole (Solea senegalensis). Aquaculture, 288: 147-150.

Bagheri T., Imanpour M R. 2011. The efficacy, physiological responses and hematology of Persian sturgeon, Acipenser persicus, to clove oil as an anesthetic agent, Turkish Journal of Fisheries and Aquatic Sciences, 11: 477-483.

Bolasina S.N. 2006. Cortisol and hematological response in Brazilian codling, Urophycis Brasiliensis (Pisces, Phycidae) subjected to anesthetic treatment. Aquaculture International, 14: 569-575.

Iversen M., Finstad B., McKinley R.S., Eliassen R.A. 2003. The efficacy of

metomidate, clove oil, Aqui-Sk and BenzoakR as anaesthetics in Atlantic salmon (Salmo salar L.) smolts, and their potential stress-reducing capacity, Aquaculture, 221: 549-566.

Sudagar M., Mohammadi-Zarejabada A., Mazandarani R., Pooralimotlagha S. 2009. The Efficiency clove powder as an anesthetic and its effects hematological parameters on Roach (Rutilus rutilus), Journal of Aquaculture Feed Science and Nutrition, 1:1-5.

Svobodova Z., Pravda D., Palackova J. 1991. Unified methods of haematological examination of fish. Research Institute of Fish Culture and Hydrobiology, Vodnany, Edition Methods, 22. 31 pp.

Evaluation about fatty acid profile Artimia decapsulated cyst and their obtained naplus in three geographic cyst for aquaculture consumption

Nemat Peykaran Mana*1, Mir I Limed Sayed Hasani1, Hadi Asadi2, Hamid Reza Pour Ali1, Naser Agh3, Ahmad nezami Baloochi1.

1 Caspian sea international research institute, Rasht. Iran.

2 M.Sc. Student - Guilan University

3 Center of Artima and aquatic animal research. Uremia- University. * Email:nemat147p@yahoo.com

Introduction

A investigation carried out about determination of Artimia profile fatty acid and their naplus obtained from 3 region of Iran (Uremia lake, Maharloo lake and Mighan lake). The research carried out on laboratories of Artimia Research Center. Most of fish fed concentrate diet but couldn't digestive and suction them. Preparation and production live food is a basic branch in animal aquatic nutrition science especially for rearing of shrimp. Not only live food has very diversity but also they have very different quality. Artimia have a great effect on survival and growth of fish and shrimp larvae ( Azari Takami , 1999) therefore. Evaluation about fatty acid profile Artimia can help for survival and growth rate in aquatic animal also effectiveness on value and price of Artimia

(Agh,2001).

Artimia is a valuable nutrient source for fish and shrimp larvae. artemia use in aquatic animal feed on different way. in aquatic animal feed Utilization of Naplus is very current, because naplus is rich about valuable nutrient (Azaritakami, 2000).also, since 1930, artimia has changed to a valuable nutrient for animal aquatic nutrition ( Agh, 2001).

However, fish can consumption concentrated diet but they can't digestion them. Preparation and production live food is very important in animal aquatic science nutrition, Because actually with out live food, culture of aquatic animal such as shrimp is unavailable

Strain of artimia without poly unstated fatty acid ( EPA) or deficient

doesn't suitable for aquaculture nutrition (Leger et al ., 1987). Genetic structure or parents nutrition variation may led to difference sensible in fatty acid composition for Strains of artimia or one strain. This phenomenon can belonged to admixture of two cases(Schauer et al., 1980) .

Notonly live food has very diversity but also they have very different quality. Artimia "a valuable nutrient" is very important in animal aquatic nutrition such as fish and shrimp. evaluation about biochemical index such as protein, lipid, isocaloric indicator, amino acid, fatty acid , pigments, pollution via poisons and other heavy metal is one of the artimia alimentary value aspects (Hossieni ghatere, 1998).

Material and Method

Preparation of samples for fatty acid analyze, encapsulate cyst and decapsulation carried out via method of Bengston et al ., 1991. also preparation of hatching naplues from decapsulated cyst carried out via method of Sorgeloos et al., 1997. After 24 h, hatching naplus we recollected and washed by water then were transfer to aven (60 oc ) for drying.

After 24h , samples were removed from aven , crunch with pounder and changed to powder form. Samples were stocked to film box and transferred to refrigerator. The samples wee stocked to refrigerator until to analyze time. Fatty acid analyze carried out by gas chromatography system ( CG).

Results

Composition of fatty acid by different decapsulated cyst and their naplues resulted for three Iranian strain artimia were evaluated separately. Kind and quantity of any fatty acid were determined by sediment time, height and surface of sub round compare with standard time for fatty acid fixation for gas chromatography system ( CG).

Limitation of standard fatty acid and limitation of sensibility system led to indiscrimination density fatty acids. Quantity of fatty acid each sample calculated by percent of fatty acid (gr fatty acid / total fatty acid).

Result of samples ( %) showed to diagram 1 and 2.

o5

■5

mm

Пп. п

□ 3jI*-

ooooooooo9766oo5oppppo-

Figure 1: the range of decapsulated artimia cyst fatty acid Obtained from three region in Iran cyst fatty acid

oooooooo ooooooooooooooooo-

^O55!6677 COOCOCOOCOooOoooooo-(i>H О—-О-1-!!!—-О          o                              o                                   o     op""^

Figure 2: the range of naplii fatty acid Obtained from three region in Iran

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oo.oo

o5.oo

J.

3

з­oo.oo

5.oo

Discussion

One of the important topics about utilization Artimia in aquaculture

such as shrimp is Artimia alimentary value. utilization and economic value Artimia determined by fatty acid especially double band high unstated fatty acid and their unit in cyst and artimia biomass (Agh,

2001).

Proportion DHA/EPA is very important for pigment structure and fish resistant against diseases. Result showed a decrease for proportion DHA/EPA in all samples. But proportion DHA/EPA in decapsulate cyst and naplii from Maharloo Artimia were 0.02%. Proportion DHA/EPA in other treatment was zero. Legger (1986) reported amount of (EPA) 20:5n-3 obtained from different artinmia cyst were different about 0.3 to 15.4% in different area. Also they found a increase amount of indispensable fatty acid 20:5(n-3) (27.0 to 14.57 %) for different races about 0.27 to 57/14 %.

Other research indicated, indispensable fatty acid (EPA and DHA) necessary for resistant and survival fish and shrimp larvae against disease .also they have major role for pigment production.. Several study indicated, artimia has low alimentary value for aquaculture larvae because amount of EPA is very low . Also artimia has a little of DHA or not exist (Sorgeloos, 1997b) .

Schauer et al (1980) reported there are very difference for EPA belonged sanfrancisco Artimia strains (0.3 -13). But amount of EPA belonged to uremia lake were lower than monitored strains in 11 and 41 appendix, however were higher than other strains. In the other hand, in this study, the most of samples had more fatty acid unstated 20:5 (n-3) about 0.27 for Arak lake aartimia cyst that were difference with Maharloo lake artimia cyst. This result agreement with other result in the world. This result showed highest fatty acid C20:5n3 (EPA) (14.57%) C22:6n3 (DHA) (0.34) and n-3HUFA (14.92) belonged to Maharloo lake artimia cyst, respectively.

Agh, N.2002. Biology and ecology of Lake Urimia Artimia. Country Scientific Council Final Report. The program of national research. Number of registration 1443. pp: 130

Azari Takami, GH.,2000. The evaluation about resistance for poly unsaturated fatty acid( EPA, DHA) during Artimia enrichment with different oil and starvation periods. Final report. Tehran university of natural resources, pp: 32.

Bruggmean, E., Sorgeloos, P. and Vanhaecke, P. 1980. Improvments in the decapsulation technique of Artemia cysts. In: The brine shrimp Artemi. Vol.3. Ecology, culturing and use in Aquaculture.persoone G., P. Sorgeloos, O. Roels and E. Jaspers (Eds), Universa press, Wetteren, Belgium. pp: 357-372 and pp: 260-268.

Hossini Ghtreh, H., 1998. evaluation abourt Artimia elementary value by determination of protein, lipid and fatty acid composition in different growth stage. PhD Thessis, Uremia University veterinarian, pp: 90.

Leger, P., Bengtson, D.A., Simpson, K.L. and Sorgeloos, P. 1987. International study on Artemia. xxxv.Thechniqes to manipulate the fatty acid profile in Artemia nauplii and the effect on its nutritional effectiveness for the marine crustacean Mysidopsis bahia (M.). In: Artemia research and its Application Vol.3. Ecology, culturing, use in Aquaculture, Sorgeloos, P., D.A., Bengtson, W. Decleir, and EJasper (Eds)Universa press, Wetteren, Belgium, pp: 411-424.

Mayer, R.j. 2002. Morphology and biometry of three populations of Artemia (Branchioopoda: Anostraca) from the Dominican Republic and Puerto Rico,

Hydrobiology, 486, pp:29-38.

Peikaran Mana, N. 2007. Quantitative and Qualitative assessment of Artemia Cysts, Decapsulated cysts and Nauplii from 3 Geographical regions of Iran, MSc thesis, Lahijan Islamic Azad University, pp.144.

Schauer, P.S., Johns, D.M., Olney, C.E. & Simpson, K.l. 1980. International study on Artemia IX. Lipid level energy content and fatty acid composition of the cysts and newly hatched nauplii from five geographical strains of Artemia, pp: 365­373.

Sorgeloos., p. 1997b. Lake urmia cooperation project contral item A: Report on the Determination Identification of Biological Characteristics of Artemia urmiana for Application in Aquacalture. pp:6-17

Vanhaecke, p. and Sorgeloos, P. 1982. International study on Artemia , XVIII. The hatching rate of Artemia cysts - a comparative study. Aquacultural Eng., 1(4):

263-273.

Effects of probiotic Lactobacillus sp. in the diet on hematological and boichimical factors of common carp (Cyprinus carpio) fingerlings

Mohsen pourabasali1*, Mitra esmaili2, Mohammad gholizadeh1, Azanin peyvandi1

1 Department of Fishery, Islamic azad University, babol branch,Iran

2 Department of Fishery, Islamic azad University, azadshahran branch, Iran

Abstract

A 60 days study was conducted to determine the effects of Lactobacillus plantarum isolated from healthy mugil cephalus, as probiotic on blood and serum parameters of common carp fingerlings.The result indicated that all probiotic treatment had better haematological parameters (RBC, total leucocytes, Hb, HCT, MCH, MCHC) performance,when compared with the control group. A significant increase (P< 0.05) in the total protein and globulin concentration of common carp serum could be found in all probiotic treatment compared with the control,while there was no significant difference indicated in albumin level (P>0.05). Lower cortisol and glucose content were observed in T3 (P<0.05) than the control. The serum lysozyme activity was significantly greater in T3 compared with the control fish (P<0.05).

Keyword: Lactobacillus sp, Blood, Serum, Cyprinus carpio

Introduction

It has been well documented that the use of antibiotics makes drug-resistant microorganisms and keeps antibiotic residues in fish flesh and environment (Aly et al., 2008). In this respect, use of probiotic bacteria is a new approach gaining acceptance in aquaculture to control potential

pathogens ( Kim and Austin 2008). Probiotics have been used in aquaculture as a means of disease control, supplementing or even in some cases replacing the use of antimicrobial compounds. Lactic acid bacteria (LAB) are known microorganisms that have probiotic properties. They can produce inhibitory compounds such as lactic acid, hydrogen peroxide, diacetyl, acetaldehyde and bacteriocin. Lactobacillus plantarum is highly versatile and found in many different ecological niches such as meat, fish, dairy products and in the GI tract (Aymerich et al., 2003). The total erythrocyte count (Irianto and Austin, 2002), total leucocyta count fed with Saccharomyces Cerviase, haemoglobin, mean corpuscular volume, haemoglobin concentration and mean corpuscular haemoglobin concentrations, total protein, albumin, globulin, albumin-globulin ratio, cortisol and glucose were reported to increase in Labio rohita provided with Bacillus subtilis as probiotics (Kumar et al., 2008). In the present study, we investigate the effects of isolated lactic acid bacteria L.plantarium as probiotic from intestine of Mugil Cephalus as probiotic supplemented on hematological and boichimacal parameters of Common Carp (Cyprinus carpio) fingerlings.

Materials and methods

2.1 experimental design:

The isolation of Lactobacilus from fingerlings of Grey Mullet (Mugil cephalus) gut were performed using the methods reported by Carnevali et al. (2004). Twelve aquaria (70 L) with three replicates for treatment and control were used. 240 C.carpio fish with mean body weight of (15 ± 1.65 g) were adjusted in three fiberglass tanks (250 L), and were fed basal diet three times daily for 2 weeks and then they were randomly dispensed in 12 aquaria at a density of 20 fish per each for 60 days culture. Fishes were fed three times daily with each feed. The experimental diet was fed at feeding rate of 3% fish weight per week. Temperature range of water was 24-26 C. The Lactobacillus sp was grown for 72 hours at 25oC in MRS agar and harvested by centrifugation (10000 rpm for 15 min). The cells were re-suspended in 100ml of saline.

Egg albumin was added and this emulsion was then applied to fish feed by mixing in a drum mixer for 15 min. The Lactobacillus was added to the diet plates in three levels (T1= 1/5x106, T2: 3x106, T3: 4/5x106 CFU/mg). The basal diet formulation is consist of: 20.5%Fish meal,14.4 %Wheat flour, 5.5% Gluten meal, 5.2 % Fish oil, 17.8 % Rice bran, 31% Soybean meal, 2.25 %Vitamin1 & mineral premix, 0.2% Vitamin C, 2.05% Dicalcium phosphate, 1.1% L-Lysine.The basal diet proximate composition is shown in Table 1.

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