Abstract
Pesticides are the major source of concern as water pollutants. Persistent organochlorines can accumulate in food chains. Chlorpyrifos (O,O -diethyl O -(3,5,6-trichloro-2-pyridinyl) phosphorothioate; CAS No. 2921-88-2; CPY). CPY is a widely used organophosphorus insecticide that is available in a granular formulation for treatment in soil. Pesticides are used to control a wide range of pests including Mosquitoes. Mosquito borne diseases infect millions of people every year globally. The aim of current study was to screen the fresh water pollutants, water quality parameter in irrigation water from El Mahmodia stream, El-Beheira Governorate, Egypt and to determine the adverse effects of Chlorpyrifos on the larvae of Culex mosquito larvae as bio-indicator. The LC95 of Chloropyrifos insecticide was 6331.30 at 24h and increased to 230506.4 after 48h of exposure to the Chloropyrifos insecticide. It is noted that the effect of the exposure time of Chloropyrifos insecticide on the LC50, LC25 and LC95 values had a synergistic interaction with time, as it increased after 48h of exposure when compared to 24 h of exposure. The 0.09 ppm concentration of Chloropyrifos had no effect on the second instar Culex larvae, as there is no mortality over time; the same result is also with the control 0 ppm. There is no effect after 72, 96h of exposure of the population to the detected insecticide. This study concerns with studying the pollutants along El Mahmodia stream in El Beheira governorate in Abo Homs city with its abundance during the four seasons (2016-2017), as well as studding the physicochemical parameters in it. Another concern of this study is estimating the effect of one of this pesticides (Chloropyrifos) insecticide on the second instar Culex mosquito larvae, determining the lethal concentration of this insecticide on the Culex larvae. Along the study area, pesticides are used within a high ratio on the agriculture scale with its four main categories organophosphates, organochlorine, pyrthoid and carbamates. Organophosphates and organochlorine are used at a wide range. Pollutants measuring achieved by using GC-MS as water samples collected seasonally and analyzed, there is a big number of Pollutants which was found as well as other compounds which are banned, such as DDT. The physicochemical parameters Turbidity, COD, BOD in El Mahmodia stream exceeded the desirable limits of (Egyptian Law 48/1982), (WHO, 1993) and (FAO, 1985) although the other parameters as EC, PH, DO,TDS TSS are to be within the permeable limits. HCO3, NH4. Cu also was found to exceed the desirable limits while, Pb, Mn, Fe and Cd within the permeable limits. Chloropyrfos as an organophosphate pesticide used in the present study which was found with 0.09 mg/l in the stream water, used to estimate its effect on the Culex mortality, determining LC25, LC50 and LC95. The experiment continued for 96 h but after 48 h there is no effect of Chloropyrfos on Culex larvae. The experiment began with 20 second instar Culex larvae immersed in 100 ppm, 10 ppm, 1 ppm, 0.1 ppm and finally 0.09 ppm of Chloropyrfos insecticide with five repeats to each concentration, it is noted that the lethal concentration increase after 48h of experiment, the larval mortality decrease with time.
Author Contributions
Copyright© 2019
E Eissa Ebrahim, et al.
License
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This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Introduction
Persistent organochlorines can accumulate in food chains. This bioaccumulation has been well documented with the pesticide dichlorodiphenyltrichloroethane (DDT) There are 76 drains discharging drainage water into Nile system with annual volume of about the half of the total drainage water Trace metals such as Zn, Cu and Fe play biochemical role in the life processes of all aquatic plants and animals. In the Egyptian irrigation system, the main source of Cu and Pb are industrial wastes, while that of Cd is the phosphatic fertilizers Chlorinated pesticides (OCPs) and polychlorinated biphenyls (PCBs) were routinely used in large quantities for agricultural and industrial purposes
Materials And Methods
Water samples were taken from El Mahmodia stream 31̊ 06̍ 16̎ N 30̊ 18̍ 52̎ E. Water samples (2.5 L) were collected in clean glass bottles at water surface and 50 cm below water surface. Water samples were collected during the period of September 2016 to August 2017 (samples were taken seasonally); Heavy metal ions were measured by using the Atomic Absorption spectrophotometer Model THERMO SCIENTIFIC ICE 3000 series AAS with hollow cathode lamp for each element being measured (Cu, Pb, Zn, Cd and Fe) according to GC-MS analysis of water: Extraction of water samples using Empore disc technology according to EPA 3535 The percentage of 48% EC chlorpyrifos (devagro kimya tarim san vetic Torkey) was used to determine the lethal concentration LC25, LC50 and LC95 against Culex larvae. Mosquitoes culture and rearing: Mosquitoes culture brought from Alexandria University faculty of Agriculture and accommodate for (2) weeks in laboratory. Bioassay of Detected Pollutant in Water The mosquito larvae were exposed to a wide range of tested concentrations to find out the activity range of the materials under test. After determining the mortality of larvae in this wide range of concentrations, a range of 5 concentrations, yielding between 10% and 95% mortality in 24 h or 48 h is used to determine LC50 and LC95 values. Batches of 20 insects at the second instar larvae were transferred by means of droppers to Petri dish each containing 20 ml of water. Small, unhealthy or damaged larvae were removed. The appropriate volume of dilution is added (20 ml) water to Petri dish to obtain the desired target dosage, starting with the 100, 10, 1, 0.1, 0.09 ppm concentration. Five replicates were set up for each concentration and an equal number of controls (5 replicates) are set up simultaneously with tap water. After 24 h exposure, larval mortality was recorded. For slow acting insecticides, 48 h reading was required. Moribund larvae are counted and added to dead larvae for percentage mortality. Dead larvae are those that cannot be induced to move when they probed with a needle in the siphon or the cervical region. Moribund larvae are those incapable of rising to the surface or not showing the characteristic diving reaction when the water is disturbed. The results are recorded to detect the LC25, LC50 and LC95 values. The form will accommodate sex separate tests of four concentrations, each of five replicate. Analyzing the data occurred by using SAS and LDP Line The first analysis examined the abundance of the physicochemical parameters and heavy metals in the water samples which collected from EL Mahmodia stream, measuring its mean ,SD and 95% SD of it. The second analysis examined the lethal concentration LC25, LC50, LC95 and X
Results
Physicochemical determination of water samples were collected in August 2017 from four locations repeats along El Mahmodia stream, Water samples were taken (about 20 cm) below the water surface to avoid floating matter. The Electrical Conductivity (Ec) was determined with a mean of 0.50 mg/l and ± 0.01 for SD ( EC: Electrical Conductivity , TDS: Total Dissolved Solids , TSS: Total Suspended Solids , TS :total solids , DO: Dissolved Oxygen , BOD: Biological Oxygen Demand, COD: Chemical Oxygen Demand, MPN : Microbiological examination. FID: Fold of increase or decrease = (detected value – reference value)/ reference value * 100. * A, FAO 1985 .,* B, law 48/1982,* C, WHO (1993) ,* D, the guidelines of WHO (Chapman, 1992). The total solids (TS) were determined with the mean of 293.3mg/l, ±27.83 for SD. The Dissolved Oxygen (DO) detected with 4.35 mg/l for its mean, SD value was ±0.1 ( (Na): sodium, (K): potassium, (Ca): calcium, (Mg): magnesium, (HCO3):bicarbonate alkalinity, (NH4):Ammonia, (PO4): Phosphorus, (NO3):Nitrate, (Fe): Iron, (Cu): Copper, Manganese, (Cd): cadmium. A, FAO 1985 .,* B, law 48/1982,* C, WHO (1993) ,* D, the guidelines of WHO (Chapman, 1992), F, USEPA, 2001. The potassium (K) determined with 7.99 mg/l for mean value. The calcium (Ca) means detected with 31.8 mg/l, SD value was±4.54. 16.38 mg/l is the value of magnesium (Mg) mean which recorded, SD value was ±5.30 ( Extraction Efficiency (Recovery tests): For assessment the efficiency of SPE approach as extraction tools for extraction the pesticide residues in water samples, the average percentage of recoveries (%Rec.) from fortified blank samples of water were determined and the percent relative standard deviation (%RSD) for recoveries were calculated. For that purpose a laboratory water blank were fortified with the mixture of OPCs to reach the final concentration of 0.1 ug and 1ug/l. Fortified water samples were extracted and analyzed as previously mentioned. Average percentage of recoveries (%Rec.) were determined and the percent relative standard deviation (%RSD) for recoveries were calculated. All data of residue analysis were corrected according to these obtained recovery percentage values. ( As shown in The detected compounds in winter stream water samples are: Decamethylcyclopentasiloxane, 1-(2-Acetoxyethyl)-3,6-diazahomoadamantan-9-one oxime, (2-(Aminoacetyl)amino-4-methylpentanoyl)amino)acetic acid, 2',6'-Dihydroxyacetophenone, bis(trimethylsilyl) ether, 4H-1-Benzopyran-4-one, 2-(3,4-dimethoxyphenyl)-3,7-dimethoxy-, 11,16-Bis(acetyloxy)-3,20-dioxopregn-4-en-21-yl acetate, 9,12-Octadecadienoic acid (Z,Z)-, 2,3-bis(trimethylsilyl)oxypropyl ester m-Dioxane, 5-(hexadecyloxy)-2-pentadecyl-, trans-, Dodecamethylcyclohexasiloxane, 2-(9-Borabicyclo3.3.1non-9-yloxy) ( The side effects on the second instar mosquito larvae: The present study had been undertaken in order to screen the pollutants, water quality parameter, and mineral content in irrigation water from El Mahmodia stream, El-Beheira Governorate, Determine the adverse effects of detected-pesticides (Chlorpyrifos) on the larvae of Culex mosquito larvae as a bio-indicator, with a serial number of Chloropyrifos concentration (100ppm, 10ppm, 1 ppm, 0.1ppm and 0.09 ppm), cross ponding to determine the lethal dose concentration LC25 ,LC50 and LC95 of cholropyrifos insecticide on Culex larvae. The treatment occurred by a serial concentration of Cholorpyrifos; 0, 0.09, 0.1,1,10,100 ppm applied on the mosquitoes larvae. After 24h, 48h mortality percentage was recorded as; at 24h, Cholorpyrifos killed 50% of the mosquito larvae population at 24.52 ppm. While at a longer time 48h, the 50% of the mosquito larvae population were killed by 755.65 ppm. After 24h, the detected concentration of Cholorpyrifos on 25% population mortality was 2.51 ppm, although it was 72.37 ppm after 48h of Cholorpyrifos exposure to 25% population of mosquitoes. It is detected that after 24h, the Cholorpyrifos killed 95% of the mosquito larvae population at 6331.30 ppm, while after 48h, the 95% of the mosquitoes larva population were killed by 230506.4 ppm. Mortality percentage were calculated using LDP line software (Ehab soft, Egypt) according to Finney 1951. Mortality percentage were calculated using LDP line software (Ehab soft, Egypt) according to Finney 1951. The concentration of 0.09 ppm had no mortality on mosquito larvae in the second 48 h to 1h. The control also not had any mortality population on mosquito larvae. It was noted that after 72 h and 96 h there was no effect on mosquitoes larvae, as the equal number of inserted larvae were constant at the end of the experiment.
Parameter
Mean
SD
95% Confidence Limits
Reference values
Ref Association
EC
0.50
± 0.01
0.006
0.043
0.31–1.87mS cm–1
A
pH
7.68
±0.05
0.03
0.1985
7.94–8.506-8.56.5 - 8.4
BCA
TDS
249
±14.16
8.02
52.81
500 mg/l
B
Turbidity
9.17
±2.66
1.51
9.94
5 NTU
D
TSS
51
±3.36
1.90
12.552
<100
B
TS
293.3
±27.83
15.76
103.8
-
-
DO
4.35
±0.1
0.05
0.3729
<5
B
BOD
23.75
±2.5
1.41
9.3214
<6-10 mg/l.
B
COD
24.75
±3.86
2.18
14.4004
<10-15 mg/l
B
MPN
12075
±3379.7
1914.6
12601.4
5000/100cm3
B
Parameter
Mean
SD
95% CL SD
Reference
Ref Association
Na
49.91
0
.
.
200mg/l
C
K
7.99
0
.
.
12 mg/l
E
Ca
31.8
±4.54
2.57
16.93
-
-
Mg
16.38
±5.30
3.004
19.77
100 mg/l
E
Cl
56.40
±3.28
1.86
12.24
(less than 200 mg/l)
B, C
HCO3
253.3
±11.71
6.63
43.66
<200 mg/l
B
NH4
1.42
±0.14
0.08
0.55
<0.5
B
NO3
5.89
±0.93
0.53
3.48
(not exceed 45 mg/l).
B
PO4
0.01
±0.004
0.002
0.01
1 mg/l
B
Fe
0.65
±0.24
0.12
0.12
(<1 mg/l)
B, C
Cu
10.91
±2.59
1.46
9.66
(<1.0 mg/l)
B,C,F
Mn
0.09
±0.01
0.007
0.08
(<0.5 mg/l)
B, C
Cd
0.002
±0.001
0.007
0.004
0.003mg/l
C
OCPs
(Rec. %) ± RSD1ug/l
Chlorpyrifos-methyl
99.3 ± 4.0
Heptachlor
92.8 ± 6.2
Dieldrin
99.1 ± 1.2
p,p-DDD
95.6 ± 1.5
p,p-DDT
102.3 ± 4.0
Methoxychlor
99.3 ± 4.0
N
Rt
COMPOUND NAME
A
W
Sp
Su
1
5.37
Decamethylcyclopentasiloxane
√
√
2
5.65
1-(2-Acetoxyethyl)-3,6-diazahomoadamantan-9-one oxime
√
√
3
5.76
Nonadecane
√
√
4
6.1
2',6'-Dihydroxyacetophenone, bis(trimethylsilyl) ether
√
√
5
6.3
4H-1-Benzopyran-4-one,2-(3,4-dimethoxyphenyl)-3,7-dimethoxy-
√
√
6
6.41
11,16-Bis(acetyloxy)-3,20-dioxopregn-4-en-21-yl acetate
√
√
7
6.57
9,12-Octadecadienoic acid (Z,Z)-, 2,3-bis(trimethylsilyl)oxypropyl ester
√
√
8
7.03
m-Dioxane, 5-(hexadecyloxy)-2-pentadecyl-, trans-
√
√
9
7.13
Dodecamethylcyclohexasiloxane
√
√
10
7.28
2-(9-Borabicyclo3.3.1non-9-yloxy)-3-((2-(9-borabicyclo(3.3.1) phenyl non-9-yloxy)ethyl)sulfanyl)propyl ether
√
√
11
7.95
Sulfurous acid, butyl hexyl ester
√
√
12
8
1-Tridecene
√
√
13
8.04
) )5LPregnane- 3,20 L diol, 14à,18à-(4-methyl-3-oxo-(1-oxa-4-azabutane-1,4-diyl))-, diacetate
√
√
14
8.21
2,7-Diphenyl-1,6-dioxopyridazino(4,5:2',3') pyrrolo(4',5'-d)pyridazine
√
√
15
9.15
Phthalic acid, butyl tetradecyl ester
√
√
√
16
10.05
Phenol, p-tert-butyl-
√
√
17
10.91
3-Hydroxyspirost-8-en-11-one
√
√
18
11.1
Hexadecamethyl-cyclooctasioxane
√
√
19
12.18
Dasycarpidan-1-methanol, acetate (ester)
√
√
√
20
13.3
Phthalic acid, isobutyl octadecyl ester
√
√
√
21
13.37
Phthalic acid, butyl 2-ethylbutyl ester
√
√
22
14.24
n-Hexadecanoic acid
√
√
√
23
14.32
1,2-Benzenedicarboxylic acid, dibutyl ester
√
√
√
24
15.95
Phenol, 3,5-bis(1,1-dimethylethyl)-
√
√
25
17.89
1,2-Benzenedicarboxylic acid, butyl phenylmethyl ester
√
√
26
18.18
2,3-Bis((trimethylsilyl)oxy) propyl (9E,12E,15E)-9,12,15-octadecatrienoate
√
√
27
19.83
6,9,12,15-Docosatetraenoic acid, methyl ester
√
√
28
19.83
Fenretinide
√
√
√
29
20.45
Methyl((24-oxo-3,7,12 tris((trimethylsilyl)oxy)cholan-24-yl)amino)acetate
√
√
30
21.22
cis-13-Eicosenoic acid
√
√
31
21.44
Propanoic acid, 2-(3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl)-
√
√
32
22.08
Cyclopropaneoctanoic acid, 2-octyl-, methyl ester
√
√
33
24
Estra-1,3,5(10)-trien-17β-ol
√
√
34
25.59
Dihydroxanthin
√
√
35
26.23
1-Hexadecanol, 2-methyl-
√
√
36
26.5
Corynan-17-ol, 18,19-didehydro-10-methoxy-, acetate (ester)
√
√
37
26.61
16-Octadecenoic acid, methyl ester
√
√
38
27.06
Pentadecanoic acid, methyl ester
√
√
39
27.12
1,2,4-Trioxolane-2-octanoic acid, 5-octyl-, methyl ester
√
√
40
31.46
Tricyclo(20.8.0.0(7,16)]triacontane, 1(22),7(16)-diepoxy-
√
√
41
32.19
9,12,15-Octadecatrienoic acid, 2,3-bis((trimethylsilyl)oxy)propyl ester, (Z,Z,Z)-
√
√
42
32.52
1H-Cyclopropa(3,4)benz(1,2-e)azulene-5,7b,9,9a-tetrol, 1a,1b,4,4a,5,7a,8,9-octahydro-3-(hydroxymethyl)-1,1,6,
√
√
43
33.24
Oleic acid, 3-(octadecyloxy)propyl ester
√
√
44
33.88
Phthalic acid, di(2-propylpentyl) ester
√
√
45
33.92
Bis(2-ethylhexyl) phthalate
√
√
√
46
34.41
Benzeneacetonitrile, α-((4-(dimethylamino)-2,5-dimethoxyphenyl)methylene)-4-nitro-
√
√
47
34.59
9-Desoxo-9-x-acetoxy-3,8,12-tri-O-acetylingol
√
√
48
35.05
Olean-12-ene-3,15,16,21,22,28-hexol, (3β,15α,16α,21β,22α)-
√
√
49
35.21
Oleic acid, eicosyl ester
√
√
50
36.81
Pregnane, 3,11,17,20,21-pentamethoxy-, (3α,5β,11β,17α,20β)-
√
√
LC95
LC25
Confidence Limits of LC50
LC50
Time
Higher
Lower
1.41
6331.30
2.51
45.65
14.78
24.52
24
1.43
230506.4
72.37
5485.96
258.10
755.65
48
Discussion
Various physicochemical parameters like temperature, pH, DO, turbidity, BOD, nitrate, phosphate, TDS, and fecal coliform were determined by following the standard methods The suspended particles are the main source of turbidity in water. In this study, the suspended solid concentration in waters a long El Mahmodia stream was reported as 51 mS/cm. In this study, the suspended solid concentrations in waters along El Mahmodia stream within the permissible limits of law 48/1982 (<100 mg/l). Total Dissolved Solids (TDS) concentration in water samples collected along El Mahmodia stream was 249 mg/l. The TDS show an increase in its values at all recommended sites. In irrigation water, the salinity hazard is related to the high values of TDS. The total dissolved salts along the El Mahmodia stream were less than 450 mg/l and there was no restriction on using it for some susceptible crops Oxygen concentration in water is very important for fish. It is worth mentioning that unpolluted waters typically have BOD values of 2 mg/l or less, whereas those receiving wastewater may have values up to 10 mg/l or more, particularly near to the point of wastewater discharge The ammonia NH3 concentration in stream water was 1.42mg/l, the concentrations exceed the desirable Limits <0.5 (Egyptian Law (48/1982). These high values may be attributed to the increased de nitrification in water, when the oxygen concentration is low. The total alkalinity (HCO3 concentrations) in water samples was 253.3 mg/l. The concentrations of HCO3 concentrations was high that can be attributed to the decomposition in the dead phytoplankton leading to the release of CO2 dissolving to water in the form of HCO3. The concentration of HCO3 measured at El Mahmodia stream exceeds the permissible limits of Egyptian Law (48/1982) which was<200 mg/l. Phosphorus is an essential nutrient element for living organisms and exists in water bodies as both dissolved and particulate forms. Natural sources of phosphorus are mainly derived from weathering processes of phosphorus bearing rocks and the decomposition of organic matter The term heavy metal refers to any metal and metalloid element that has a relatively high density ranging from 3.5 to 7 g/cm3 and is toxic or poisonous at low concentrations Most of the organic pollutants detected at the peaks in GC-MS data analysis were identified as endocrine disrupting phthalate esters, fatty acids, phenolic acids, carcinogens, and aquatic toxicants, plasticisers, which are classified as priority pollutants due to their severe toxicity in living being Dihydroxybenzoic acid might be raised in El Mahmodia stream water as a key metabolite of biodegradation of polyaromatic hydrocarbons (PAHs) during wastewater treatment It is estimated that the total amount of reused treated wastewater in Egypt was about 1.4 billion m In the present study, bioassays were carried out to evaluate the insecticidal concentration of chlorpyrifos on the second instar Culex larvae. Surveys in Egypt date back to 1903. According to these surveys eighteen culicine and eleven anopheline species have been encountered in the different parts of Egypt. The mean level of physicochemical parameters and heavy metals as Turbidity, BOD, COD, NH4, HCO3, MPN, Cu and physicochemical parameters which determined showed an increase in its values compared to the standard safety criteria of the Egyptian Law (48/1982), the guideline of WHO Organochlorine pesticides, prohibited since the early 1980s, are still detectable in the environment. Organophosphates are found in high rate in the stream, Chloropyrifos is an Organophosphate pesticides found at concentration of 0.09 m/l in the stream water. Effect of the exposure time of Chloropyrifos insecticide on the LC50, LC25 and LC95 values had a synergistic interactions with time as it increased after 48h of exposure when compared to 24 h of exposure. The 0.09 ppm concentration of Chloropyrifos had no effect on the second instar Culex larvae, as there is no mortality. Also there is no effect on mosquito mortality after 72h and 96h of exposure to the detected concentration of Chloropyrifos insecticide.