In this study, the toxicity of MTBE on the crustacean Daphnia magna (Cladocera) has been studied. At the beginning, for finding the range of sensitivity of D. magna to MTBE, a test had been conducted at 24h. The initial and final tests with synthetic MTBE has been done with containing solutions of water accommodated fractions (WAFs) in periods of 24 and 48h and then static bioassay was calculated in two phases of tests and the data were obtained and analyzed by SPSS 13 by using probit analysis. In 24h, lc10, lc50 and lc90 for MTBE on D. magna were 345, 646 and 941mg/l and for 48h, they were 361, 476 and 893mg/l, respectively. Also, the maximum allowable concentration (MAC) of MTBE (WAFs) on D. magna was determined in 24 and 48h, 64.6 and 47.6mg/l, respectively. Results indicated that D. magna could be used for toxicity tracing MTBE in surface and ground water, because these crustaceans have perfect resistance against pollution of MTBE. It is also concluded that after 24 and 48h exposure time, there was no significant difference observed in the activity and mortality of D. magna

Jaafar Zadeh, N.

Khodadady, M.

Piri, M.

Ranjbar, M.

Aquatic Commons

 Iranian Journal of Fisheries Sciences                                                               10(3) 541-545                     2011         
Survey of Methyl Tertiary Butyl Ether (MTBE) toxicity using 
bioassay on Daphnia magna 
 
Ranjbar M.
1*
; JaafarZadeh N.
2
; Piri M.
3
 ;  Khodadady M.
4 
 
Received: January 2010                   Accepted: May 2010 
 
1- Payam Nour University, Iran. Department, Payame Noor University,19395-4697 Tehran, 
I.R. Iran. 
2 - School of Public Health, Medical Sciences Faculty of Jondishapour University, Ahwaz, 
Iran. 
3- Inland Waters Aquatics Stocks Research Center, P.O. Box: 139 Gorgan, Iran. 
4 – Science and Research Branch of Islamic Azad University, Ahwaz, Iran. 
*Corresponding author’s email: Email:ranjbar_45522@yahoo.com 
 
Keywords: Bioassay, MTBE, Daphnia magna, Water pollution 
In this study, the toxicity of MTBE on the 
crustacean Daphnia magna (Cladocera) 
has been studied.  At the beginning, for 
finding the range of sensitivity of D. 
magna to MTBE, a test had been 
conducted at 24h. The initial and final tests 
with synthetic MTBE has been done with 
containing solutions of water 
accommodated fractions (WAFs) in 
periods of 24 and 48h and then static 
bioassay was calculated in two phases of 
tests and the data were obtained and 
analyzed by SPSS 13 by using probit 
analysis. In 24h, lc10, lc50 and lc90 for 
MTBE on D. magna were 345, 646 and 
941mg/l and for 48h, they were 361, 476 
and 893mg/l, respectively. Also, the 
maximum allowable concentration (MAC) 
of MTBE (WAFs) on D. magna was 
determined in 24 and 48h, 64.6 and 
47.6mg/l, respectively. Results indicated 
that D. magna could be used for toxicity 
tracing MTBE in surface and ground 
water, because these Crustaceans have 
perfect resistance against pollution of 
MTBE. It is also concluded that after 24 
and 48h exposure time, there was no 
significant difference observed in the 
activity and mortality of D. magna.  
Since Methyl Tertiary Butyl Ether 
(MTBE) was used widespread as a fuel 
oxygenated in gasoline, it commonly 
contaminates ground water, drinking 
water, coastal waters, and other aquatic 
bodies (Werner et al., 2001). 
Ecotoxicologic studies of MTBE were 
mainly performed in freshwater ecosystem 
and reported the concentration of acute 
toxicity, ranging from 136 to 1000mg/L 
which varied according to organism 
species. Therefore, the toxicological 
studies mixture of chemicals is growing 
(Hernondo et al., 2002). The main goal of 
this study was to evaluate toxic effects of 
MTBE using Crustacean (D. magna). The 
second goal of this research included 
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  542    Ranjbar et al., Survey of Methyl Tertiary Butyl Ether (MTBE) toxicity………. 
assigning of (MAC) of MTBE and 10% 
concentration of this material in water. 
Static bioassay was calculated in two 
phases of examination and the data 
obtained were analyzed by SPSS 13 using 
probit analysis.  
In this stage, the toxicity test has been 
done on D. magna using water 
accommodated fractions prepared in the 
laboratory in 24 and 48h. The results 
showed that in 24h, mortality of D. magna 
begun with concentration of 310mg/L and 
in 48h mortality ended with concentration 
of 30mg/L and 497mg/L. The results 
indicated in which concentrations the 
mortality was zero and in which 
concentrations mortality was 100 percent. 
At the end, by entering these 
concentrations into SPSS software, data 
were analyses in probit model. The 
analyzation of data was preformed by 
using ANOVA and effect values of MTBE 
on D. magna with use of variance ratio 
(F), chi-square (x
2
) and values of 
regression coefficient (Tables 1 to 4).  
Data on toxicity of MTBE suggest that 
this compound has a low toxicity towards 
aquatic organisms. Acceptable data on the 
toxicity of MTBE to freshwater organisms 
were available for three species of fish, six 
species of aquatic invertebrates, and three 
species of plants. For example at 
concentrations of 388-2600mg/l, MTBE is 
toxic to vertebrates (Rousch & 
Sommerfeld, 1998). Acute toxicity values 
(96h LC50) for the rainbow trout 
(Oncorhynchus mykiss) (Hocket, 1997) 
and bluegill sunfish (Lepomis 
macrochirus) ranged from 887mg/l to 
1054mg/l (Hocket, 1999). The 96h LC50 
estimate for juvenile fathead minnows 
(Pimephales promelas) was 980mg/l. Also 
the toxicity value of MTBE on trout fish 
had been studied and the obtained results 
showed that 96h Lc50 value for this fish 
was 752mg/l (Beigy, 2006). On the other 
hand, MTBE in water and soil of gasoil 
which pumps around Tehran had also been 
studied by Ardalani (2003).  
Acute toxicity values for invertebrates, 
ranged from 340mg/l (48h LC50) for 
Ceriodaphnia dabia (Hocket, 1997) to 
1742mg/l (48h E50 survival) for 
Chironomus tentans (Hocket, 1999). 
Depending on the time of exposure, 
MTBE is a toxic to invertebrates at a 
concentration of 57 to >1000mg/l (Werner 
et al., 2001). EC50 values of 7.4-41.8mg/l 
have been reported for acute toxicity of 
MTBE towards bacteria such as 
Salmonella tyhimurium and 
Photobacterium phosphoreum (Kado et 
al., 1998). In microalgae, MTBE is toxic at 
the concentration of 184-4800mg/l, when 
tested with species such as Selenastrum 
capricornutum and Synechococus 
leopoliensis (Werner et al., 2001). 
MTBE’s toxicity on D. magna 
(EC50=720mg/l) is content with a reported 
96h EC30 measurement of 681mg/l 
(Hernando et al., 2002). Other researchers 
have claimed that MTBE is not toxic to D. 
magna at the concentration of 1-1000mg/l 
in exposures up to 48h (Gupta & Lin, 
1995). These results, obtained with the 
static mode of the D. magna assay are 
explicable by volatilization of MTBE. 
Volatilization can greatly reduce the 
concentration of MTBE seen by the test 
organisms over duration of the assay 
(Hernando et al., 2002). In conclusion, 
MTBE alone is toxic to the tested species 
at concentrations that are significantly 
greater than the levels of pollutants 
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                                                     Iranian Journal of Fisheries Sciences, 10(3), 2011                                        543  
  
commonly detected in the environment 
(Hernondo et al., 2002).  
In this research, we used standard 
Checkousluaky (No. 46 6807) (Asobeda  
& Ukuosa, 1995) (Table 9) in limitation 
between 100 to 1000 which showed 
MTBE had low toxicity on D. magna and 
also MTBE did not have significant effects 
on the organism's mortality. These 
crustaceans have perfect resistance against 
pollution of MTBE and the results 
indicated that D. magna could be used for 
toxicity tracing MTBE in surface and 
ground water. Therefore, MTBE that was 
determined after 48h exposure (according 
to standard of Checkousluaky) had a low 
toxicity. 
On the whole, the animals are sensitive to 
pollution in all seasons and this depends 
on metabolic activities and physiological 
manners of them. So, different species 
have different sensitivities regarding water 
accommodated fractions (WAFs) 
(Shahlapour, 1992). Therefore, MTBE and 
concentration of pollution in certain times 
of the year may cause mortality for a 
species of  animals and does not have any 
effect on the others. Generally circular 
forms forms Shows more toxicity to chain 
forms (Piri and Falahi, 1998). The other 
case is toxicity of some refined since we 
can't generalize the results yield of finding 
with the MTBE to other oil products at all.  
 
Acknowledgments 
We would like to appreciate and express 
our gratitude to the Official Ahvaz Water 
and Waste Central Laboratory which 
generously cooperated with us in preparing 
this research.. It would be appreciated for 
help of  Mr. Rabiae and respected 
employees of this center for their helps and 
other staffs. 
 
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                                                     Iranian Journal of Fisheries Sciences, 10(3), 2011                                        545  
  
  
 
Table 1:  ANOVA for data of logarithmic (11) in 24h 
  
 
 
 
 
 
Table 2:  ANOVA for data five treat in 48h 
 
 
 
 
 
Table 3: Coefficient for data of logarithmic (11) in 24h 
 
 
 
    
 
Table 4: Coefficients for data of five treat in 48h 
 
 
 
 
 
 
 
       
     
Model 
Sum of 
Squares df Mean Square F Sig. 
Regression 14,826 1 14,826 1,793 ,189 
Residual 281,174 34 8,270 - - 
Total 296,000 35 - - - 
Model 
Sum of 
Squares df Mean Square F Sig. 
Regression 1,342 1 1,342 ,351 ,562 
Residual 61,158 16 3,822 - - 
Total 62,500 17 - - - 
Model 
Unstandardized 
Coefficients 
Standardized 
Coefficients 
T sig - - 
- B Std. Error Beta - - 
Case Sequence ,062 ,046 ,224 1,339 ,189 
(Constant) ,190 ,979 - ,195 ,847 
Model 
Unstandardized 
Coefficients 
Standardized 
Coefficients T Sig. 
 - B Std. Error Beta -  -  
Case Sequence ,053 ,089 ,147 ,593 ,562 
(Constant) ,667 ,961 - ,693 ,498 
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Short communication: Survey of Methyl Tertiary Butyl Ether (MTBE) toxicity using bioassay on Daphnia magna

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Short communication: Survey of Methyl Tertiary Butyl Ether (MTBE) toxicity using bioassay on Daphnia magna

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