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2 Department of Natural Resources, Isfahan University of Technology, Isfahan 84156-83111, Iran. E.mail firstname.lastname@example.org, email@example.com
In this study early cichlid larvae (5-weeks-old, 15.8 mm length and 110 mg weight) and advanced larvae (10-weeks-old, 25.6 mm length and 240 mg weight) consumed C. quadrangula at range 220-584 ind. day1 larvae-1 and 5281956 ind. day-1 larvae-1, respectively, as well as suitable growth and survival rate. The study demonstrated that C. quadrangula could be used as live feed purposes for larval rearing in aquaculture.
Keywords: Freshwater Ceriodaphnia quadrangula, Mayan cichlid larvae, ornamental fish larvae, ingestion rate
Use of this species as live food is very suitable for aquarium or ornamental fish larvae, such as Mayan cichlid, Cichlasoma urophthalmus. This species belongs to Cichlidae family. It has interesting colors, with its eight black bands and its large ocellus on the caudal peduncle which gives it its scientific name. It is native to the Atlantic slope of tropical Mesoamerica (Central America) and inhabits lakes, rivers, rocky shorelines, lagoons, estuaries, coastal islands and mangroves. It inhabits waters with temperatures from 18 to 34 °C and salinity from 0 to 40 ppt.
This species is a dietary generalist, consuming organisms from a variety of disparate taxa, especially crustacean zooplankton. This paper aims to mass culture of C. quadrangula and its consumption by an ornamental fish, Mayan cichlid (Cichlasoma urophthalmus) larvae.
Materials and methods
Mass Production of C. quadrangula to Rear Mayan Cichlid Larvae
Ceriodaphnia quadrangula was cultured for two months in three plastic tanks (50 + 3 L) containing of EPA medium with initial density of 30 ind. L-1. Green microalgae, Scenedesmus quadricauda was added at a concentration of 4-6 x 105 cells mL-1 every other day depending on C. quadrangula population size and growth of S. quadricauda in the cultures. Additional set of C. quadrangula was estimated by counting the total number of population and were processed for feeding of Mayan cichlid larvae. After live harvesting of C. quadrangula individuals, ingestion rate of two groups of Mayan cichlid (different in age, size and weight, Table 1) were determined using different density of C. quadrangula. The experiment was carried out with cichlid larvae which were obtained from aquarium shop located in Isfahan, Iran. The cichlid larvae stock were acclimated to laboratory conditions of filtered autoclaved freshwater, 26 + 1 °C temperature and 12h: 12h light : dark for 72 hours and fed on artificial feed which use in aquarium shop.
The cichlid larvae density in each beaker was 5 larvae L-1. Each beaker was stocked with 10 larvae. Aeration and uniform distribution of C. quadrangula was provided. Each feeding treatment was replicated thrice. Three control beakers (without larvae) containing only C. quadrangula were used for each concentration. The fish larvae were transferred daily to new container with the same volume of water and prey density. C. quadrangula counting and the required number for experiment were performed in a zooplankton chamber (Bogorov's plate chamber). The prey density in both control and feeding treatments (with larvae) was measured daily from at least 5 sub-samples. Ingestion rates (IR) and weight specific ingestion (WSI) of Mayan cichlid larvae were calculated according to Farhadian et al. (2007).
Table 1- Mean (± S.E, N=10) total length, width, wet weight, and dry weight of cichlid
Cichlasoma urophthalmus larvae.
Data were analyzed using two-way analysis of variance (ANOVA). Differences in treatment means were compared by Duncan's multiple range tests. The maximum population specific growth rates (SGR) were transformed to Arcsine square root to ensure a normal distribution (Zar, 1984) and tested for statistical significance by two-way ANOVA. All statistical analyses were carried out using statistical package of SPSS
(SPSS, version 2002).
Mass production of C. -q1uadrangula in plastic tank (50 L) had a density of 2000 ± 235 ind. L-1 after 20 day culture fed on S. quadricauda. Results showed that early cichlid fish larvae consumed C. quadrangula at 220-584 ind. day-1. Similarly, advance cichlid fish larvae ingested C. quadrangula at 528-1956 ind. day1. This study indicated that cichlid larvae, at early and advance stage, can ingest C. quadrangula. The WSI significantly increased with increasing prey density (P<0.05, Fig 5). The WSI values ranged from 2.99 to 7.95 % for while for early fish larvae, these values ranged from 3.95 to 14.62 % for advance larvae per day at different food concentrations (Fig 1). The larvae dry weights in each larval group were significantly higher at 10 ind. mL-1 than 2 and 5 ind. mL-1 (Table 2). The maximum survival rate and dry weight were observed at 10-weeks larvae at prey density of 10 ind. mL-1.
Table 2- Dry weight (nig/larvae) and survival rate (%) of cichlid Cichlasoma urophthalmus larvae fed on Ceriodaphnia quadrangula in 10 days period at three prey densities (ind.mL-1).
Prey d amity
90 3 c
96 6 "
86 6 "
Values are mean ± standard error from three replications. Values within each row do not share the same superscript are significantly different (P<0.05)
Figure 1 Mean (±SE, N=3) ingestion rates (A) and weight specific ingestion (WSI) (B) of Mayan cichlid larvae fed on different Ceriodaphnia quadrangula density. Bars in the same letters are not significantly different (P>0.01).
In this study, cichlid larvae ingested C. quadrangula in both larval sizes (Table 2, Fig. 1) due to its suitable body size and absence of spine in the caudal parts. Some species of zooplankton did not consume by fish larvae due to spines in body (Zaret, 1980). Compared results between 5-weeks-old and 10-weeks-old larvae showed that ingestion rate 10-weeks-old larvae were increased. These differences could be related to improvement of prey capture and handling skills of 10-weeks-old cichlid larvae, therefore they consume higher numbers with increasing prey density (Alanis et al., 2009).
Authors are grateful to Isfahan University of Technology (IUT), Isfahan, Iran to support this study.
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Investigating the salinity potential of Persian Sturgeon, Acipenser persicus during early developmental stages
M. Mardaneh Khatooni*1, B. Mojazi Amiri1, A. Mirvaghefi1
1* Department of Fisheries and Environmental Sciences, Faculty of Natural Resources, University of Tehran, 31585-4314 Karaj, Iran, Email:maryamkhatooni85 @ gmail.com
The factors which affect fish eggs mortality were divided in environmental and internal factors (Heath, 1992). Some studies have investigated the environmental factors impacts on abnormalities and morality during early developmental stages of sturgeon (Dettlaff and Ginsburg, 1992; Ruban et al, 2006). Although Salinity effects have been studied extensively in marine fish embryos and larvae (Yang and Chen, 2006; J0rgensen and Hansen, 2010), but they are limited in freshwater species (Albert et al, 2004; Bonislawska, 2009). It has shown that some freshwater teleost eggs can be incubated and hatched in 5ppt salinity but fertility and eggs hatching rate of freshwater teleost decreases in saline water (Gbulubo and Erondu, 1998; Fashina-Bombata and Busari, 2003). Holliday and Jones (1967) found that the egg salinity resistance was lowest in Blastula and gastrula in freshwater teleost eggs and Tylore (1971) found the most sensitivity just enclosing blastopore prior to hatch. However, there is limited information on how salinity affects developmental stages of sturgeons (Jian-Yi et al., 2006).
Materials and methods
the fertilized eggs were divided into 50g groups (and transferred directly into recycling incubators that designed for this study with 2, 4, 6, 8ppt salinity treatments. Trial duration at the end of larvae hatching was 7 days at 18°C. Mortality and abnormality percent were distinguished and determined according to Dettlaff and Ginsburg (1992) method during three developmental stages of gastrula, s-type heart formation and before hatch stages during exposure to the different salinity treatments.
Result has shown that the effect of salinity on abnormality percent was different in each developmental stage. For example the abnormality percent increased significantly in gastrula stage only in 6 and 8ppt treatments. The abnormality rate in s- type heart formation stage was significantly higher in all treatments compare to control but it was still below 20%. The abnormality rate increased significantly in 4 and 6ppt treatments before hatch stage. Also our result showed that the salinity significantly affect abnormality percent during developmental stages of each treatment (P< 0.05). these abnormalities were included some disruption in cell segmentation, retreated developing embryos and the yolk abnormal shortening in 6 and 8ppt in gastrula stage. The hatching rate was recorded 84.19±3.2% in 4ppt treatment, while it was recorded 64.03%±1.8 in 6ppt treatments. Hatching did not occur in 8, 10 and
Comparing the results of salinity effects on each stages of all (0.5, 2, 4, 6 and 8ppt) studied treatments showed that the embryos of A. persicus were more sensitive to salinity in gastrula stage in higher salinities like 6 and 8ppt, but they are more tolerable in gastrula stage as the abnormality and mortality rate were higher in heart formation and before hatch stage in lower salinity (2 and 4ppt). Zotin (1965) reported that the water absorption will cease in gastrula stage and it will start to absorb rapidly in heart formation stage again in Beluga (Huso huso). However, Holliday and Jones (1967) found that the egg salinity resistance was lowest in blastula and gastrula in freshwater teleost eggs and Tylore (1971) found the most sensitivity just enclosing blastopore prior to hatch. Sawant, et al (2001) also reported increasing salinity potential along with developmental stage in zebra fish in low salinity as 2ppt (from embryonic cleavage up to gastrula stage) and cytological examinations indicated that higher salinity mainly impaired the nuclear division of the embryonic cells. Others reported some less salinity potential in fresh water embryos
during incubation period. For example more than 2ppt in zebrafish (Sawant et al, 2001), 4.8ppt in White fish (Coregonus lavaretus)(Albert et al, 2004) and 3ppt in Sea trout (Salmo trutta L.) (Bonislawska, 2009) was recorded critical to hatch. Consequently Our result showed the more salinity tolerance in low salinity range in gastrula stage in Persian sturgeon but this potential decreased along with increase in salinity more than 6ppt.
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The effects of oligofructose on haematological parameters and survival rates of Acipenser stellatus juvenile
Mahsa khoshbavar Rostami1, Hoseinali Khoshbavar Rostami2, Mohammad Ahmadi3
1 Department of Fisheries, Islamic Azad university of Qhaemshahr, Iran
2 Iranian fisheries research organization, inland waters research center of Gorgan, Iran
3 Head of Islamic Azad University, Qaemshahr Branch, Iran
This study investigates the effects of oligofructose on some haematological factors and survival rates of Acipenser stellatus juvenile. The study included 3 treatment repeated in triplicates. Fish (29±3 g) were supplied and stocked at density of 10 fish in tanks and fed on experimental diets for 76 days. Our results showed that prebtioic positielt affect white blood cells and neutrophils while other haematological parameters remained unaffected. The highest neutrophil and white blood cells observed in 1% treatment. Also fish fed control diet had highest survival rate, 96.67±5.77. these results showedthat oligofructose affect the components of innate immune system and subsequently alter disease resistance. Thus, we suggest administration of this dose in Acipenser stellatus diet.
Keywords: oligofructose, prebiotic, haematological factors, survival, Acipenser stellatus
Sturgeons are commercially valuable species, which are currently highly endangered due to over fishing, loss of habitat and decrease of water quality (Carmona et al. 2009). for stock enhancement beside artificial
propagation, culture up to marketable size and brood stock culture should be considered (Ustaoglu et al., 2004). Fish welfare and survival are the two important consideration in aquaculture which depends on physiological status of fish (Flikinger et al., 2003). Administration of dietary supplements like pro and prebiotics suggested for improvement of fish health (Hoseinifar & Pooramini,...). Prebiotics, defined as non-digestible food ingredients which beneficially affect the host by selectively stimulating the growth of and/or activity of health-promoting bacteria in the intestinal tract (Gibson et al. 2004). Previous studies on administration of dietary inulin and oligofructose confirmed increase of production, survival and health status of Siberian sturgeon (Mahious & Olevier, 2005), turbot (Mahious et al., 2006), Beluga (Akrami, 2009; Hoseinifar et al., 2011), Indian white shrimp (Hoseinifar et al., 2010) and Caspian roach (Khosravi, 2011). However, there is no information available on the effects of oligofructose as prebiotic on Acipenser stellatus. Haematological parameters are proper tools for determination of fish health status (Hoaston, 2006). However, possible effects of dietary prebiotics on various aspects of fish physiology remained to be investigated.
Thus this study investigates the effects of oligofructose on some haematological factors and survival rates of Acipenser stellatus
Material and methods
This study performed in Ghare Soo research station for 76 days. Acipenser stellatus (average weight of 29±3 g), obtained from the International research Institute of Sturgeons (Guilan province, Iran), were randomly stocked into 12 tanks at a density of 10 fish per tank (3 tanks per treatment). The prebiotic used in this study, oligofructose, was produced by Sigma and used 1 & 2 % in diet. Commercial diet (Biomar) milled in order to produce a fine powder. After addition of prebiotic levels, Warm water was added to the milled basal mixture and grinded to produce 2 mm pellets. The pelleted diets were stored in plastic bags at 4 °C for further use. At the end of feeding trial blood samples (2cc) from 3 fish per tank were taken from the caudal vein for haematological analysis. Whole blood was suspended in diluent (Natt and Herrick 1952) for erythrocyte and total leukocytes counts (WBC) using a haemocytometer. Haematocrit was
determined by the microhaematocrit method (Brown, 1988). Haemoglobin levels were estimated using Sahli's method (Blaxhall and Daisley 1973). Mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were calculated according to the standard formula.
Survival rates were conducted based on following formula: Sr=(Nt-No)x100 Water quality parameters were measured daily using WTW device (Germany). After checking for normality and homogeneity of variance data were subjected to a one-way analysis of variance (ANOVA). When significant differences were observed, Duncan's multiple range tests were performed (Zar 1994).
The results of this study showed that oligofructose had no significant effects on Hb, red blood cells count, MCV, MCH, MCHC. However, fish fed prebiotic had significantly higher Neutrophils and WBC when compared to control (P < 0.05). The highest values observed in 1% treatment. Moreover, the highest survival rate was observed in control treatments (96.67±5.77) while fish fed 2% prebiotic had lowest survival rate (83.33+15.28) (Fig 1).
Table 1. Haematological parameters of Acipenser stellatus fed different levels of dietary oligofructose
226.88 +13.26 a
45.89 +1.64 a
19.38 +3.43 a
0.9 +0.24 a
0.96 +0.03 a
(mm3 x106) RBC
20.67+ 4.58 a
25.22+ 3.17 a
4.11+ 1.53 a
4.49+ 0.19 a
( g/dl) Hb
Table2. Differential count of WBC of Acipenser stellatus fed different levels of dietary oligofructose
8844.44 +302.46 b
81.11 +7.04 a
94.78 +1.64 a
80.67 +16.65 a
2.44 +0.69 a
Our results revealed that oligofructose had no effects on survival rates of Acipenser stellatus. Similar results reported about inulin effects on beluga (Akrami, 2009). Likewise, inulin had no effects on trout survival rate (Akrami et al., 2007). In addition, MOS negatively affect survival rate of Nile tilapia (Vendemiatti et al., 2003). However, Hoseinifar (2007) reported that oligofructose significantly increased beluga survival. Prebiotics effects on fish survival are depending on ability of intestinal microbiota of fish gut for fermentation of prebiotics (Hoseinifar & Zare, 2010). Fermentability of prebiotics is depends on degree of polymerization. Inability of Prebiotics fermentation and accumulation of prebiotic will results in negative effects on fish physiology (Olsen et al.,
The haematological parameters of fish are reported to be effected by a range of factors which include species, size, age, physiological status, environmental conditions and dietary regime (e.g. quality and quantity of food (Dumeizan et al., 1997). Our results confirmed that prebiotic had no significant effects on haematological parameters. However, fish fed 1% dietary oligofructose had higher Htc, Hb. The highest WBC count was observed in 1% treatment was significantly higher that other treatments. AKrami reported that the highest WBC count in beluga fed 1% prebiotic treatments. Elevation of WBC count can be considered as a positive effect on immune system (AKrami et al., 2007). Similarly, Hoseinifar (2007) reported that although RBC count remained unaffected, WBC
count was significantly higher in 2% treatment.
However, it has been reported that prebiotic had no effect on fish survival. Administration of 0.5 and 2% inulin in trout diet had no significant effects on haematological parameters include RBC, Htc, Hb (Sheikholeslami et al., 2007).
Differential count revealed that neutrophil was significantly higher in 1% treatments. However, Hoseinifar (2007) reported that prebiotic didn't affect Neutrophil while significantly increased lymphocytes amount. Similarly, inulin significantly increased lymphocyte amounts in beluga (Akrami et al., 2007). The results of the present study hint at a possible immunomodulatory effect; however, additional studies are required to focus on the effects of oligofructose on the immune response and disease resistance of Acipenser stellatus.
Special thanks to Dr. Rostami head of inland water research center of Gorgan, Dr. Ahmadi head of Islamic Azad University, Savadkooh branch, staff of Gharehsoo research station, Mr. Iri and Molarami for their helps during the trial
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The effect of diazinon on osmoregulation capability in Caspian roach (Rutilus rutilus) fingerlings
Kheyrollah Khosravi Katuli1*, Bagher Mojazi Amiri1, Smaeel Smaeelzadegan1, Saeed Yelghi2
1 Department of Fisheries, Faculty of Natural Resource, University of Tehran, Karaj, 31585-4314 Iran
2 Iranian Fisheries Research Organization (IFRO), Inland Waters Fisheries Research, Gorgan, Iran
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The Caspian roach (Rutilus rutilus) is an indigenous, economically important and a main food sources for sturgeon species in the Caspian Sea (Coad, 1980; Keyvanshokooh and Kalbassi, 2006). This species is considered at the risk due to overfishing, water pollution, and loss of habitat and spawning sites (Kiabi et al., 1999). Therefor Iranian Fisheries organization attempted to semi-synthetic propagation and restock of juveniles to the estuary of Qare Soo River located in northern Iran.
Diazinon [O, O-diethyl O-(2-isopropyl-6-methylpyrimdin-4-yl) phosphorothioate], is an organophosphate pesticide and is being used extensively to control rice pests. High levels of diazinon were reported in the Qare soo rivers estuaries (Shayeghi et al., 2006). Thus we investigated the effect of sublethal diazinon exposure on osmoregulatory capabilities of caspian roach fingerlings. Fresh water conditions during the early stages of the life cycle of anadromous fish can have a significant impact on survival and the successful return of spawning adults in marine environment.. The effect of diazinon exposure on the changes of physiological indices of Caspian roach fingerlings were testified. Active ion uptake in the gills takes place by means of chloride cells. These highly specialized cells are affected by many kinds of pollutants, such as pesticides (Wendelaar Bonga and Lock, 1991). Electrolytes are involved in osmotic activity and provide buffer systems and mechanisms for the regulations of pH (acid-base balance). These parameters were also measured to determine the ability of
fingerlings to osmoregulation activity in sea water after exposure to the diazinon pesticide.
Caspian roach fingerlings (average weight of 0.770 ± 0.05 g), used in this experiment originated from the Sijowal Caspian sea teleost fish reproduction center (Golestan province, Iran), and were acclimatized for at least 1 week under laboratory conditions. Amount of pesticide that used in this experiment was close to amount of this pesticide in river and selected according to 96 h LC50 value 0.0 (control), 1.0, 2.0 and 3.0 mg L-1 of diazinon.