Electrolyte Analysis of drinking water of Djamaa (Algeria) region and study its effect on the kidneys in Wistar rats
Samir Derouiche, Taissir Cheradid, Messaouda Guessoum
Faculty of Natural and Life Sciences, El-Oued University, El Oued 39000, El Oued, Algeria
*Corresponding Author E-mail: dersamebio@gmail.com
ABSTRACT:
The aim of the current study is to evaluated the effect of drinking Water in Djamaa region on kidney function in rats. For this purpose, female Wistar rats divided into three groups (n=5), the first group of rats received mineral water serve as control, the second group of rabbits received tap water with (TDW) and the third group of rats received filtered water (FDW). Received drinking water for 60 days. Qualitative analysis of water showed a high level of chloride, calcium, potassium and sodium in TDW and low in FDW of Djamaa region as compared to mineral water. Our results showed a significant increase (p<0.05) of serum urea, creatinine and sodium levels and decrease in potassium and chloride in TDW and FDW group as compared to control. results obtained about hematological markers shows a significant decrease of Red blood cell (p<0.001), Hemoglobin (p<0.01), White blood cells (WBC) (p<0.001) and platelet (p<0.001) levels in TDW group compared to control. Results represent revealed also a significant increase (p<0.05) in MDA level and decreased (p<0.05) in antioxidant parameters such as GSH concentration, GST, SOD and catalase activities in TDW group as compared to control. In conclusion, this study showed that imbalance of electrolyte levels in tap water or filtered water is a major cause of kidney damage.
KEYWORDS: Tap water; filtered water; kidney; stress oxidative; rats.
INTRODUCTION:
Chronic kidney disease (CKD) is increasingly recognized as a global public health problem [1]. During the past three decades, the incidence and prevalence of end stage renal disease (ESRD) have risen progressively. For example, annual new cases of ESRD increased from approximately 14,500 in 1978 to 100,359 in 2002; during the same period, the number of individuals on dialysis and with kidney transplants increased from 42,000 to 431,000 [2].
There is a wide spectrum of kidney disease, which can be rapid onset (acute) or longer term (chronic) [3]. In order to identify the risk factors of renal disease, An individual's genetic and phenotypic make up puts him/her at risk for kidney disease factors such as smoking, exposure to heavy metals, excessive alcohol consumption and the use of analgesic medication [4]. Moreover, CVD, hypertension, diabetes and obesity are traditional risk factors [5]. In addition, environmental pollution including air and water contamination causer or aggravates many acute and chronic human disease [6]. Indeed Drinking water pollution is a relatively new problem and increases the stress arising as a result of unprecedented population growth, urbanization, and industrialization since 1990s [7,8]. Many of recent landmarks in scientific research have shown that in human beings, Oxidative stress is an important factor causing metabolic and physiological alterations and various diseases in the body [9], it is as a consequence of increase a reactive oxygen species and decrease in antioxidant defenses in prevalent in many health problems like CKD [10]. The aim of this study is to estimate the role of water used in drinking in a Djamaa (Algeria) region as a risk factor in developing kidney problems.
MATERIALS AND METHODS:
Chemicals:
Sodium chloride (NaCl), Hydrochlorid acid (Hcl), Hydrogene peroxide (H2O2), Thiobarbituric acid (TBA), Methanol, Coomassie Blue, Butylate dihydroxy toluene (BHT), Trichloroacetic acid (TCA), Phosphate-buffered (KH2PO4, K2HPO4), Ethylene diamin tetra-acetic acid (EDTA) were of analytical grade
Qualitative study of Djamaa drinking water:
Samples collection:
to obtained some comparatives information of drinking water quality, a snapshot study was done on 3 drinking water samples of 29 samples divided between 3 mineral water, 5 tap water (TW) and 4 filtered water (FW) which obtained from deferent regions in Djamaa according to an orderly sampling method. This experimental study was conducted in the ADE laboratory in El Oued Unity
Animals and Experimental design:
Females rats with weight (181.74± 6.96) were bought from animals service of Pasteur institute in Algeria, they are installed in faculty SNV, University of El oued, Algeria in plastic cages divided in three groups of 5 rats of each. They kept in the animals breeding house for adaptation. The animals were adapted to laboratory condition photoperiod (12 h of night/12h of darkness), an ambient temperature of 22 ± 03°C and humidity of (63.2 ± 14%) for two weeks. The standard diet and water are free for the animals during period of adaptation. The realization of the experimental part is respect to the ethical approval.
Females rats were randomly divided into the following three groups (5 rats):
Group I: female rats were given Mineral water (control group).
Group II (TDW): the female rats were given Tap drinking water for 60 days.
Group III (FDW): the female rats were given filtered drinking water for 60 days
filtered drinking water contained "Hypersperse 700-11" which is a viscous solution as the intercalate product. this product is used in the demineralization station of the water during the filtration stages to avoid the precipitation of chlorine in the tubes of the installation.
Blood collection and tissue Preparation:
At the end of 8th week of experiment the animals were fasted for 16 h, anesthetized by chloroform inhalation then sacrificed by decapitation. The blood was collected in EDTA tubes for hematological analysis. The serum was obtained by blood centrifuging at 3000rpm for 10 min and frozen at 20°C until the use for urea, creatinine, calcium and electrolytes levels assay. Then, the kidney of rats of different groups was rapidly excised, weighed and stored at- 20°C until use for oxidative stress evaluation.
Determination of biochemical and hematological markers:
Serum urea, creatinine, calcium parameters levels were determined by autoanalysis (BIOLIS24j) use commercial kit from spinreact, spain (ref: urea-20141, creatinine-20151, calcium-20051). Determination of the ionogram parameter (Sodium, potassium and chlorine) by Automatic electrolyte analyzer (Easylute). Hematological analysis (FNS) is performed by the hematology autoanalyzer (Sysmex).
Water electrolytes and Conductivity measurement:
In this study we used Flame atomic absorption spectrometry for the sodium and potassium measurement in drinking water. Titrimetric method for the determination of chlorides dissolved in water with AgNO3 and K2CrO4 (Mohr's method, ISO 9297 – NA 6917) and of the calcium by Sodium Hydroxide and EDTA (1984 (F). Calcium Assay - EDTA Titrimetric Method (ISO 6058). A specifies spectrometric method for phenantroline for the determination of iron in water (ISO 6332). It is measured by electric method to the conductivity of water determination where the results are displayed directly by the conductimeter an µS with the temperature measured (conductivity, NA 749).
Oxidative markers measurement:
The method of malondialdehyde (MDA) assay is based on the reaction between the carbonyl compounds of malondialdehyde with thiobarbituric acid according the method of (Yagi,1976) [11]. The level of reduced Glutathion (GSH) was determined according the Weak and Cory (1988) [12] by measuring the optical density results from the foemation of 2-nitro-5-mercocapturic acid from the reduction of dithio-bis-2-nitrobenzoic acid, which is called Ellman reagent with SH groups exist in GSH. Gluthation transferase (GST) activity was measured spectrophotometrically based on the formation kinetics of a complex between a GST substrate: 1-chloro-2-4-dinitrobenzene (CDNB) and GSH according to thethe method of (Habig et al,1974) [13]. The assay method of superoxide dismutase (SOD) activity using the NBT by the superoxide anion (O2.), is used as a basis for detecting of presence of SOD by measuring the absorbance at 560 nm [14]. The catalase activity consists in measuring the catalase-induced loss of H2O2 contained in the sample by measuring the absorbance of H2O2 at 560 nm using a UV /visible spectrophotometer according the method of Aebi 1984 [15].
Statistical Analysis:
The statistical evaluation is carried out by the student's T test using Minitab 17.1 statistical package and the Excel 16.0 (Microsoft). The value are given as mean and standard deviations (ES) for three groups. Statistical significance was defined as P < 0.05.
RESULTS:
Qualitative analysis of water:
In our experimental analysis of water (figure 1) we noticed a high level of chloride, calcium, potassium and sodium in tap water of Djamaa region as compared to mineral water. On the other hand very few levels of calcium, sodium, potassium, chloride and electrical conductivity was recorded in filtered drinking water than mineral and tap water. In addition a low iron concentration we found in tap and filtered drinking water than mineral water.
Figure 1: Minerals, electrolytes levels and electrical conductivity of mineral water, tap water and filtered water.
Biochemical markers:
Concerning the biochemical markers (table 1), results obtained show that a significant increase (p<0.05) of urea and creatinine levels and a decrease of urea/creatinine report in TDW and FDW groups compared to control except FDW group there is no significant change in urea concentration compared to control. (table 2). Regarding the electrolytes levels, our results showed a significant elevation (p<0.05) in serum sodium level in TDW group against control and a significant decrease (p<0.05) in serum potassium and chlorine levels in TDW and FDW groups as compared to controls, But there is no significant changes (p>0.05) in serum sodium level in FDW group than the controls.
Table 1: Serum biochemical markers and electrolytes levels of rats groups studies.
FDW (n=5) |
TDW (n=5) |
Control (n=5) |
Parameters |
0.39 ± 0.03* |
0.45 ± 0.02** |
0.35 ± 0.01 |
Serum Urea (g/l) |
3.69 ± 0.42* |
6.30 ± 0.12*** |
2.64 ± 0.43 |
Serum Creatinine (mg/l) |
0.01 ± 0.010*** |
0.07 ± 0.002*** |
0.170 ± 0.02 |
Serum Urea/creat (mg/l) |
93.90 ± 2.20 |
99.75 ± 0.45* |
92.60 ± 3.10 |
Serum Sodium (mmol/l) |
56.46 ± 0.89* |
53.87± 0.10*** |
58.9 ± 1.14 |
Serum Potassium (mmol/l) |
100.05± 0.11*** |
104.75± 0.78** |
108.05 ± 0.81 |
Serum Chloride (mmol/l) |
The results are presented by mean± SEM. Significance from control, *(p<0.05), **(p<0.01), ***(p<0.001)
Hematological markers:
results obtained in table 2 shows a significant decrease of Red blood cell (RBC) (p<0.001), Hemoglobin (p<0.01), White blood cells (WBC) (p<0.001) and platelet (p<0.001) levels in TDW group compared to control. However, in FDW group the WBC and platelet levels are decreased (p<0.001) compared to control but no significant variation concerning erythrocytes line.
Table 2: Hematological parameters of blood in control and experimental rats groups.
FDW (n=5) |
TDW (n=5) |
Control (n=5) |
Parameter |
679.0 ± 17.2 |
692.50 ± 2.17*** |
717.00 ± 3.67 |
Red blood cells (104/l) |
13.90 ± 0.38 |
13.95 ± 0.05** |
14.15 ± 0.12 |
Hemoglobin (g/dl) |
32.20 ± 0.89 |
34.30 ± 0.13 |
34.15 ± 0.150 |
Hematocrit (%) |
865.7 ± 24.0*** |
794.0 ± 36.7*** |
1202 ± 205 |
platelet (103/l) |
3.87 ± 0.33*** |
5.35 ± 0.02*** |
6.60 ± 0.20 |
White blood cells (103/l) |
The results are presented by mean± SEM. Significance from control, **(p<0.01), ***(p<0.001)
Oxidative stress markers:
Our results (table 3) show that a significant increase (p< 0.001) of MDA level and decrease (p<0.05) of GSH concentration and GST and SOD activities in TDW and FDW groups compared to control. For catalase activity our results show that there was a significant increase (p< 0.05) only in FDW group as compared to control group.
Table 3: Oxidative stress markers in kidney of control, TDW and FDW groups.
Parameter |
Control (n=5) |
TDW (n=5) |
FDW (n=5) |
Kidney MDA (nmol/mg) |
299.5 ± 20.8 |
394.5 ± 19.5** |
345.33 ± 3.36*** |
Kidney GSH (nmol/mg prot) |
0.49 ± 0.27 |
0.13 ± 0.02* |
0.21 ± 0.03** |
Kidney SOD (U/mg prot) |
1.22 ± 0.13 |
0.57 ± 0.03*** |
0.60 ± 0.06*** |
Kidney Catalase (UI/g prot) |
0.57 ± 0.04 |
0.46 ± 0.004*** |
1.01 ± 0.19 |
Kidney GST (nM/min/mg prot) |
6.91 ± 0.95 |
3.45 ± 0.20*** |
5.09 ± 0.21*** |
The results are presented by mean± SEM. Significance from control, *(p<0.05), **(p<0.01), ***(p<0.001)
DISCUSSION:
Our results showed a high level of chloride, calcium, potassium and sodium in tap water of Djamaa region as compared to mineral water. As study of Zobeidi et al (2013) [16] the calcium levels show high concentration exceeding potable standards (204 to 260mg/l), it is the main urinary risk for calcium stone. Extremely hard water (hardness > 500mg/l) is also unfit for consumption because the constituent mineral such as Ca+ can depsit inside the body if present in high amounts leading to kidney stone [17] where the Calcium stone is most common type of kidney stone which occurs in about 70% -80% of case . they are usually composed of Ca+ [18]. In fact, High sodium intake may also exert direct effects on CKD progression, independent of blood pressure [19]. Sodium could also affect endothelial function via TGF-β1which play an important role in the progression of CKD [20]. In addition, Potassium plays a crucial role for sufficient functioning of the heart [21]. High potassium is a risk factor of CVM, arrhythmia and heart failure [22]. As recording to Zobeidi et al (2013) [16], the measured chloride concentration exceed 250mg/l WHO approval, with values obtained at an average of 320 to 1264mg/l. Conductivity reflects the mineralization of water [23] and has the higher level of ionic concentration activity due to excessive dissolve solid [24]. In addition, we obtained for the Iron is a very low level in the tap and filter water with ranging from 0 to 0.065 mg/l . As recording to WHO (1996) [25] the health concentration of iron in the water should be no more than 0.3 mg/l. The iron is an essential element in the human body where it installs oxygen transport proteins such as hemoglobin or myoglobin [26]. Our results showed a significant increase of serum urea, creatinine and sodium levels and decrease in potassium and chloride in TDW and FDW group as compared to control. Urea is the waste product made in the body breaks down proteins [27] and creatinine is product of dephosphorylation of creatine during the muscle contraction and filtred by proximal tubule and secreted out by kidney in urine [28], the loss of kidney filtrations declines of GFR and less excretion of wastes results an accumulation of urea and creatinine in blood [29]. The results of animal study indicate a low level the HB, RBC, Platelet and WBC in TDW groups compared to control. The study of Jeffery L.M (2013) [30] show that hemoglobin is the most iron-containing protein in the red blood cells. Hemoglobin is a protein with the property of fixing, transporting and delivering oxygen [31]. There fore anemia is characteristic of iron deficiency when hemoglobin and RBC induced anemia may cause renal dysfunction caused by low oxygen delivery, which exacerbates renal dysplasia which in turn leads to interstitial renal injury and fibrosis [32]. Our study showed a significant change in oxidative stress markers where the quality of drinking water has a major influence on public health and polong exposure to contaminated water has been known to increase the risk of disorders in kidney [33]. Our results represent a significant increase in MDA level and decreased in antioxidant parameters such as GSH concentration, GST, SOD and catalase activities in TDW group as compared to control. these results are in agreement with results of Inkielewicz. et al (2008) [34] that found The concentration of GSH and CAT, SOD activities are decreased in kidney of experimental animals exposed to some toxic products in their drinking water. Glutathione (GSH) is an antioxidant that contributes to the non-enzymatic defense system against free radical-induced oxidative stress [35]. Moreover, other toxic products of drinking water (disinfectants) may induce to oxidative stress in mammalian cells and occure intracellular imbalance more susceptible to oxidative damage [36]. In addition ,The increase in conductivity and richness in inorganic ion (K+, K+2, Na+ and Mg+2 in Tap water influences metal bioavailability and toxicity and water chemistry parameters strongly influence toxicity of metals to organisms [37] where the ionicity in drinking water by deferent electrolytes is one of the risk factors which cause CKD [38].
CONCLUSION:
This study shows the negative effects of the quality of drinking water on the kidneys due to the oxidative stress caused in rats. From the study, we can also conclude that imbalance of electrolyte levels in tap water or filtered water is a major cause of kidney damage.
ACKNOWLEDGEMENT:
This work was supported by the research project D01N01UN390120190001 funded by the ministry of higher education, Algeria and by Directorate General for Scientific Research and Technological Development.
CONFLICT OF INTEREST:
All authors have approved the manuscript with no conflict of interest.
REFERENCES:
1. Levey AS, Atkins R, Coresh J, Cohen EP, Collins EJ, Eckardt KU, Nahas ME, Jaber BL, Jadoul M, Levin A, Powe NR, Rossert J, Wheeler DC, Lameire N, Eknoyan G. Chronic kidney disease as a global public health problem: Approaches and initiatives – a position statement from Kidney Disease Improving Global Outcomes . Kidney International 2007; 2: 247–259.
2. Anton CS, Michael ME, Thomas HH, Kathy HR, Dolph C, William MM, David GW, Frank V. Chronic Kidney Disease: A Public Health Problem That Needs a Public Health Action Plan. Public Health Research, Practice and Policy 2006; 3(2).
3. Naqvi R, Mubarak M, Ahmed E, Akhtar F, Bhatti S, Naqvi A, Rizvi A. Spectrum of glomerular diseases causing acute kidney injury; 25 years' experience from a single center. J Renal Inj Prev. 2015 ;4(4):113-6.
4. Kazancioğlu R. Risk factors for chronic kidney disease: an update. Kidney Int Suppl 2013; 3(4):368-371.
5. Ibrahim AG, Ahmed H, Awdah, MA. Assessment of Risk Factors for Chronic Kidney Disease in Saudi Arabia. International Journal of Science and Research 2014; 3(7): 446-450.
6. Sunil JW. Effect of water hardness on Non-communicable diseases, including chronic kidney disease of multifactorial origin (CKDmfo/CKDuo). Journal of Environment and Health Sciences, 2016; 2(1), 1-11.
7. Chen, J. Analysis of water environment in the Xinjiang arid region. Arid Environ. Monitor, 2002; 16: 223-227.
8. Velea T, Gherghe L, Predica V, Krebs R. Heavy metal contamination in thevicinity of an industrial area near Bucharest. Environ. Sci. Pollut. Res, 2009; 16: 27-32.
9. Derouiche S, Degachi O, Gharbi K. Phytochemistry analysis and modulatory activity of Portulacae oleracea and Aquilaria malaccensis extracts against High-fructose and high-fat diet induced immune cells alteration and heart lipid peroxidation in Rats. International Research Journal of Biological Sciences. 2019; 8(4): 6-11.
10. Atieh M, Mohsen N, Zahra S. Oxidative Stress in Chronic Kidney Disease. Iranian journal of kidney diseases (IJKD), 2015; 9: 165-79.
11. Yagi K. Simple Fluorometric Assay for lipoperoxyde in blood plasma. Biochemical. Medecine, 1976; 15: 212-216.
12. Weak Beker G, Cory JG Ribonucleotide reductase activity and growth of glutathione-depled mous leihemia L1210 cells in vitro. cancer letters, 1988; 40: 257-264.
13. William HH, Michael JP, William BJ. Glutathione S-transferase, The first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry 1974; 249(22): 7130-9.
14. Beauchamp C, Fridovich I. Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Analytical Biochemistry, 1971; 44(1): 276-287.
15. Aebi H. Catalase in vitro. In Methods in enzymology 1984; 105: 121-126.
16. Zobeidi A, Leila M. Physico-chemical quality of drinking water in the south of Algeria (Case of El-Oued region) study of excess minerals. International Letters of Chemistry, Physics and Astronomy, 2013; 11: 38-43.
17. Mahajan RK, Walia TPS, Lark BS, Sumanjit. Analysis of physical and chemical parameters of bottled drinking water, International Journal of Environmental Health Research 2006; 16(2): 89-98.
18. Edgar VL, Sanjay P, Elizabeth ALR, Coralie TC. Comprehensive Information About Prevention and Treatment of Kidney Disease. Book: Complete Guide for Kidney Patients (Save your Kidneys); 2eme Eds. (2015); 1-246.
19. Wang Y, Sun L, Wang H, Li X. Importance and benefits of dietary sodium restriction in the management of chronic kidney disease patients: experience from a single Chinese center. Int Urol Nephrol, 2012; 44: 549–556.
20. Ying WZ, Sanders PW. Dietary salt modulates renal production of transforming growth factor-b1 in rats. Am J Physiol, 1998; 274: 635–641.
21. Palmer BF, Clegg DJ. Physiology and pathophysiology of potassium homeostasis. Adv Physiol Educ, 2016; 40(4): 480–90.
22. Liesa KH, Dana CM, Wolfgang K, Prudence RC, Hermann B, Ben S. Association of Abnormal Serum Potassium Levels with Arrhythmias and Cardiovascular Mortality: a Systematic Review and Meta-Analysis of Observational Studies. Cardiovascular Drugs and Therapy 2018. Doi: 10.1007/s10557-018-6783-0.
23. Benrabah S, Attoui B, Hannouche M. Characterization of groundwater quality destined for drinking water supply of Khenchela City (eastern Algeria). Journal of Water and Land Development 2016. DOI: 10.1515/jwld-2016-0016.
24. Yirdaw M, Bamlaku A. Drinking water quality assessment and its effects on residents health in Wondo genet campus, Ethiopia. Environmental System Research 2016; 5(1).
25. WHO, Guidelines for drinking-water quality, 2nd ed. Vol. 2 World Health Organization, Geneva. 1996.
26. Dowell LR. Minerals in Animal and Human Nutrition. Amsterdam: Elsevier Science, 2ndEds.660 2003.
27. Weiner ID, Mitch WE, Sands JM. Urea and Ammonia Metabolism and the Control of Renal Nitrogen Excretion. Clin J Am Soc Nephrol. 2015; 10(8):1444-58.
28. Andrew CS, Naing LH, David AC, Robert LB. A Surviving Patient with Record High Creatinine. Open Journal of Nephrology 2013; 3: 217-219.
29. Pizzorno J. The Kidney Dysfunction Epidemic, Part 1: Causes. Integr Med (Encinitas) 2015; 14(6): 8-13. PMID: 26807064; PMCID: PMC4718206.
30. Jeffery LM. Iron Deficiency Anemia: A common and Curable Disease. Cold Spring Herb Perspect Med. 2013.Doi:10.1101/cshperspect. a011866.
31. Derouiche S, Serouti A, Rezzag mohcen O S. Risk of Metribuzin (Triazinone herbicide) on hematological and renal structure and function of pregnancy rabbits. Asian J Biol Sci 2019; 12:192-198.
32. Mehdi U, Robert DT. Anemia, Diabetes, and Chronic kidney Disease. Diabetes Care 2009; 32(7).
33. Wasana K, Orathai T, Surapon T. Association Between Elevated Arsenic Exposure with Chronic Kidney Disease and Oxidative Stress in Subjects of the Contamination Area. International Journal of Toxicological and Pharmacological Research, 2016; 8(3): 173-178.
34. Inkielewicz I, Czarnowski W, Gdańsk P. Oxidative stress parameters in rats exposed to fluoride and aspirin. Research report Fluoride 2008; 41(1): 76–82.
35. Derouiche S, Atoussi N, Guediri S. Assessment of Hematological Parameters, Enzymes Activities, and Oxidative Stress Markers in Salivary and Blood of Algerian Breast Cancer Patients Receiving Chemotherapy. Biochem Tech 2019; (4): 50-58.
36. Jing Y, Hui L, Li-Hong Z, Ya-Lin Z, Wen-Qing L. Oxidative stress and DNA damage induced by a drinking-water chlorination disinfection byproduct 3-chloro-4-(dichloromethyl) -5-hydroxy-2(5H)-furanone (MX) in mice. Mutation Research 2006; 609: 129–136.
37. Oluwafikemi TI, June CS, Megan JB, Annette EV, Jan GM, Lyndy JM. Generation of reactive oxygen species in relevant cell lines as a bio-indicator of oxidative effects caused by acid mine water. Water SA 2017; 43(1).
38. Kumari M, Rathnayake R, Kendaragama K, Gunarathna M, Nirmanee K. Drinking water quality in chronic kidney disease unknown Aetiology (CKDu). Int'l Journal of Advances in Agricultural and Environmental Engg. (IJAAEE) 2016; 3(1): 57-60.
Received on 28.03.2020 Modified on 12.04.2020
Accepted on 29.04.2020 ©AJRC All right reserved
Asian J. Research Chem. 2020; 13(3):219-224.
DOI: 10.5958/0974-4150.2020.00042.5