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Unorganized lead acid battery workers are exposed to lead. It causes the most occupational heavy metal toxicity. High lead affects several organs and systems of the body. Activated carbon fabric respiratory mask is known to reduce absorption of lead. The study was designed to see use of activated carbon fabric respiratory masks (ACF) in reducing lead-induced nephrotoxic effects among workers. A total of 36 male subjects aged 20 to 65 years were included in this study. Blood lead, renal markers, calcium, phosphorous, sodium, and potassium were investigated. Increased blood lead (P < 0.001), significantly decreases after using ACF mask. Renal function markers showed significant alteration in serum creatinine (P < 0.01), urea (P < 0.01), and uric acid (P < 0.001), in the study group after using ACF respiratory mask. Significant improvement in the mean levels of sodium (P < 0.0001), potassium (P < 0.05), calcium (P < 0.0001), and phosphorous (P < 0.001) after using ACF mask. Increase in generalized aminoaciduria was seen in battery workers before using ACF mask, but there was no decrease in excretion of amino acids after using ACF mask. The symptoms of lead acid battery workers did not significantly decrease after using ACF mask. Present study concludes that significant change in renal function parameters using ACF mask regularly indicates decreasing absorption of lead.
Department of Biochemistry, Krishna Institute of Medical Sciences Krishna Vishwa Vidyappeth “deemed to be university” Karad, India
Arun Patil
Department of Biochemistry, Krishna Institute of Medical Sciences Krishna Vishwa Vidyappeth “deemed to be university” Karad, India
Mandakini Kshirsagar
Department of Biochemistry, Krishna Institute of Medical Sciences Krishna Vishwa Vidyappeth “deemed to be university” Karad, India
*Address all correspondence to: ajyotsna20@yahoo.com
1. Introduction
Lead is a naturally occurring malleable and corrosion-resistant heavy metal. It can be used in building materials, automobile spray paints as protective coatings, acid storage batteries, and gasoline additives [1, 2].
It is used in lead acid battery factories in various processing steps like breaking, smelting, recycling, old batteries, and making new acid batteries. Extensive use of lead causes environmental contamination, and significant public health problems [3].
Toxic effects of lead have been known for centuries. Occupational exposure to lead results in poisoning be it asymptomatic or clinically symptomatic [4].
Clinical signs of lead toxicity are decreased libido, abdominal cramps, anorexia, nausea, constipation, diarrhea, restlessness, fatigue, irritability, sleep disturbance, headache, and difficulty concentrating. Symptoms like tremors, toxic hepatitis, or acute gouty arthritis are observed occasionally. Severity of symptoms worsens with increasing blood lead levels.
Increased lead level affects several organs of the body. Renal toxicity in humans has been recognized in earlier studies [5, 6, 7, 8]. Acute lead poisoning has been observed as disruption of proximal tubular architecture, with disturbances in proximal tubular function [9, 10, 11, 12], and manifested as Fanconi syndrome-like signs observed by glycosuria, aminoaciduria, and phosphaturia [13, 14, 15, 16, 17].
The renal manifestations of acute lead poisoning are usually reversible after chelation therapy and cessation of lead exposure [10, 18]. But for chelation therapy, it is very important to select dosage of chelating agent.
It has been observed in earlier studies that lead absorption is reduced by using activated carbon fabric respiratory mask [19, 20].
We have carefully chosen method for reduction of blood lead from absorption which is non-invasive and easy to apply. In the present study, we have provided activated carbon fabric mask to the lead acid battery workers and observed that it is useful in decreasing renal toxicity.
This is interventional study. We had planned for pre and poststudy, hence control subjects were not included.
The study selected with aim to reduce renal toxicity due to lead by the intervention of using protective gears, which suits battery manufacturing workers more feasible and comfortable.
This study comprises 36 male workers of lead acid battery manufacturing factory in the Western Maharashtra, India. The selected workers were nonalcoholic, nonsmoker, and of age range 20–45 years. Subjects on medications were excluded from the study. By using questionnaire and interview, the demographic, occupational, clinical data, socioeconomic status, dietary intake, and food habits were collected. The written informed consent from all participants was collected in regional language.
The study objectives and health hazards of lead exposure and its toxicity were informed.
2.2 Ethics declaration
Protocol committee and institutional ethics committee approval were taken prior to enrollment of study participants and as per the 1964 Helsinki Declaration [21].
2.3 Strategies for study
Lead acid battery workers were selected from unorganized lead acid battery manufacturing factories from Karad MIDC, Maharashtra, India. These workers are exposed to lead while smelting and plating of batteries.
Activated carbon fabric mask was provided to all lead acid battery workers and guided them about the use of this mask. The regular use of activated carbon fabric mask was ensured through weekly telephonic communication with all these battery workers. The workers were trained to clean the masks (recharge) every week. ACF can be easily recharged by opening the valves and filter should be removed and dipped in boiling water for 5–10 minutes and dry under the sun. This instruction was given by the authorities of Environ Products Ltd.
The activated carbon fabric mask is a respiratory full-face mask with activated carbon fabric device, which acts as barrier against chemical and biological impurities.
A unique property of an ACF fabric is the possibility to “reactivate” the fabric when it has become saturated so that it can be reused. Weak bonds are formed by physical adsorption mechanism between pollutants and activated carbon fabric surface at low temperatures. Providing heat energy, pollutant molecules can be removed from the activated carbon fabric surface [19, 20, 22]. This mask has a superior quality activated carbon fabric, which has excellent adsorbent property, removes environmental toxin, and atmospheric pollutant.
Earlier effectiveness of use of ACF masks in reducing blood lead levels of battery workers was studied for 1 month [19, 22]. Hence, we planned the study protocol for short duration, that is, for 2 months.
Blood sample was collected from battery workers. Then instructed them to use ACF masks for two months regularly. After 2 months, blood sample was collected and biochemical parameters were studied and statistically analyzed.
2.4 Biochemical laboratory tests
2.4.1 Blood lead
Ten ml blood was collected in tube containing heparin and EDTA for estimation of lead and various biochemical parameters.
Blood lead was estimated by using lead care II blood lead analyzer (Magellan Diagnostics, USA). The principle of this instrument is based on Anodic Stripping Voltammetry (ASV) to measure the blood lead level. Red blood cells (RBC) are lysed by using lead care treatment reagent, which releases the lead from RBC. To accumulate lead atoms on the test electrode, a negative potential was applied to the sensor. For releasing the lead ions, the potential was rapidly reversed, which generated the current and it was directly proportional to the amount of lead in the blood sample [23].
2.4.2 Kidney function tests
Kidney function tests were estimated by using EM360—Transasia fully automated biochemistry analyzer.
Blood urea was measured by Glutamate Dehydrogenase (GLDH) method. The rate of decrease in absorbance measured at 340 nm [24, 25, 26].
Serum uric acid was measured by Uricase-Peroxidase (POD) method. Uric acid is oxidized to allantoin by uricase with the production of H2O2. The absorbance of quinoneimine dye at 546 nm is proportional to uric acid concentration in the sample [27, 28, 29].
Serum creatinine was estimated by enzymatic method. The absorbance of produced complex at 546 nm is proportional to the creatinine concentration in the sample [30, 31, 32].
2.4.3 Minerals in blood
Serum calcium was measured by Arsenazo method. The absorbance of which is measured at 650 nm and is proportional to calcium concentration [33, 34].
Serum phosphorous was measured by UV molybdate method. The formation of reduced phosphomolybdate is measured at 340 nm and is directly proportional to the concentration of inorganic phosphorous [35, 36].
2.4.4 Serum electrolytes
Serum electrolytes like serum sodium and serum potassium are measured by method of ISE theory. Erba XL 640 fully auto analyzer is an automated, microprocessor-controlled analytic analyzer that uses ISE (Ion Selective Electrode) technology for the direct measurement of ions [37].
2.4.5 Thin layer chromatography on urine
Urine samples of subjects were collected in dark amber color bottle. Amino acids were detected in urine by using thin layer chromatography technique. Precoated cellulose sheets and butanol: acetic acid: water solvent system used. In total, 0.2 ml of urine in eppendorf tube was collected and 1.0 ml of methanol was added and vortexed, centrifuged. Supernatant was used for chromatography. A total of 5ul of supernatant was spotted on TLC sheet. TLC sheet was then kept for activation at 60°C for 10 min. TLC plate was then placed in solvent tank and allowed to ascend to 2 cm from the top of thin layer sheet. TLC sheet was removed and allowed to dry at room temperature for 10–15 min. Ninhydrin reagent sprayed over the TLC sheet and allowed to dry at room temperature. The plate was transferred to a hot air oven set at 100°C. Purple colored spots on TLC sheet were observed. These spots are amino acid spots, detected and compared with Rf values from reference values of standard in literature and also with normal urine sample [38].
2.5 Statistical analysis
Statistical comparison of blood lead, renal function markers and serum calcium, phosphorous, sodium, and potassium of lead acid battery workers were done after using activated carbon fabric mask. Unpaired “t” test applied using Instat GraphPad software. Two tail p value was calculated and was set to P < 0.05 statistically significant.
Activated carbon fabric (ACF) respiratory masks were provided to lead acid battery workers. We ensured that they were using it regularly in telephonic communication.
Table 1 shows that there is decrease in blood lead level significantly (p = 0.001) after using ACF mask.
Sr. No.
Biochemical parameters
Before using ACF masks(N = 36)
After using ACF masks(N = 36)
P value
Percentage change
1
Blood Lead Level(μg/dl)
88.31 ± 18.16 (47.6–130.20)
74.39. ± 20.16** (36.7–126.98)
0.001
−15.76
2
Blood urea (mg/dl)
26.77 ± 6.70 (16.8–47.80)
22.98 ± 6.27* (12.60–36.80)
0.015
−14.16
3
Serum uric acid (mg/dl)
7.21 ± 1.59 (3.90–10.9)
6.13 ± 1.48** (2.80–8.90)
0.0037
−14.98
4
Serum creatinine (mg/dl)
1.15 ± 0.27 (0.76–2.38)
0.99 ± 0.26* (0.60–2.10)
0.011
−13.91
5
Serum calcium (mg/dl)
8.29 ± 0.35 (7.60–8.80)
8.98 ± 0.36*** (8.07–9.70)
P < 0.0001
7.36
6
Serum phosphorous (mg/dl)
2.998 ± 0.37 (2.27–3.89)
3.59 ± 0.68*** (2.27–5.51)
P < 0.0001
19.75
7
Serum sodium (mEq/L)
139.57 ± 4.28 (124.53–145.30)
129.82 ± 3.76*** (121.85–139.78)
P < 0.0001
−6.99
8
Serum potassium (mEq/L)
3.08 ± 0.43 (2.50–4.31)
3.44 ± 0.49** (2.69–5.15)
0.0012
11.69
Table 1.
Blood Lead level, renal functions, serum calcium, phosphorous, sodium, and potassium of lead acid battery workers before and after using ACF masks.
Values in parenthesis shows reference ranges, * P < 0.05, ** P < 0.01, ***P < 0.001, figures indicate mean and SD, values in parenthesis shows minimum and maximum range.
Renal function markers like blood urea, serum uric acid, and serum creatinine decrease significantly after using ACF mask (p = 0.015, p = 0.0037, p = 0.011). Minerals like calcium and phosphorous increase significantly after using ACF mask (p < 0.0001, p < 0.0001). Electrolytes like sodium decreases (p < 0.0001) and potassium increases (p = 0.0012) significantly after using ACF mask.
Thin layer chromatography for amino acid in urine shows increased excretion of general amino acids like glycine, alanine, cysteine, histidine, leucine, isoleucine, etc. in urine as compared to normal urine sample shown in Figure 1.
Figure 1.
Thin layer chromatography for aminoacids in urine of lead acid battery workers.
From lead acid battery workers, symptoms were observed. These were muscle cramps, irritability, lethargy abdominal discomfort, and are categorized on the basis of blood lead.
Figure 2 shows mild symptoms (Adult PbB level > 60 mg/dl) like, muscle pains, prickly itchy feeling, mild fatigue, aggressiveness, irritability, lethargy, abdominal discomfort, and severe symptoms (PbB level = 70–80 mg/dl), joint pain, general fatigue, poor concentration, tremor, headache, abdominal pain, and constipation.
Figure 2.
Percentage of mild and severe symptoms of lead acid battery workers.
In this study, mainly unorganized sector of lead acid battery manufacturing workers were enrolled and they were involved in smelting, recycling, and plating of lead batteries. These workers were highly exposed to lead, which resulted in increased blood lead levels. In our earlier study, we observed that increased blood lead affects liver and kidney functions [39, 40].
Mainly lead enters the body through inhalation and ingestion of contaminated food at the site of workplace. Therefore, we have provided activated carbon fabric mask to these workers and requested all these workers to use this special mask regularly for 2 months. We have ensured that all workers were using this mask regularly by telephonically talking every week to these workers and owners of industries. Blood lead level (p < 0.01, −15.76%) was significantly decreased after using 2 months of activated carbon fabric masks, which clearly indicates that the absorption of lead decreases (Table 1, Figure 1). It is observed that the regular use of activated carbon fabric mask is really useful to reduce the blood lead levels.
Earlier study found that significant increase in serum uric acid, serum creatinine, and serum urea levels in lead-exposed Korean workers [41]. Also in our recent study, we found there is an increased level of renal parameters [39].
Urea is the marker of the kidney function, concentration of which in the plasma tends to increase in impaired kidney state.
Accumulation of lead (Pb) in the kidney increases serum urea. This could be due to impairment of glomerular membrane [1]. After using ACF mask, we observed that there was a decrease in (−14.16%) in urea level. Using regular ACF mask could decrease lead absorption, which may prevent impairment of glomerular basement membrane.
Increased plasma accumulation of uric acid may be due to deposition of lead in the proximal tubules of the nephron. These effects of lead (Pb) can subsequently lead to gouty arthritis [42]. It is observed that use of ACF mask for 2 months decreases serum uric acid (−14.98%). Uric acid is considered one of the antioxidants in plasma [43]. In this study after using ACF mask, decrease in uric acid level may spare the antioxidant property of uric acid.
In our study, we observed that high levels of serum creatinine level in unorganized battery workers decreased (−13.91%) after use of ACF mask, which is beneficial for reducing renal toxicity.
Lead inhibits the 1-α- hydroxylase enzyme in renal tubules resulting in decreased calcitriol formation [44]. Decreased calcitriol synthesis results in reduced calcium-binding proteins, which impair the calcium absorption across small intestine. Lead toxicity also causes prolonged hyperphosphaturia and hypophosphatemia in children could result in bone demineralization and rickets [15]. As mentioned in literature, same mechanism may be observed in adults and may result in osteopororsis. In our study, decreased serum calcium and phosphorous, increased by 7.36 and 19.75%, respectively, indicates ACF mask is useful in improving calcium level. Use of ACF mask decreases absorption of lead and may lead to increase in the activity of 1-α- hydroxylase enzyme resulting in synthesis of calcium-binding proteins, which have role in absorption of calcium and phosphorous from the intestine. Use of ACF mask may reduce excretion of phosphates in urine.
The effect of lead toxicity on kidney is disruption of proximal tubular architecture. This results in disturbances in proximal tubular function with laboratory evidence [9, 10, 11, 12]. Rats’ exposure to lead shows that statistically significant difference in serum mean levels of Na+, K+, and Cl [45].
We also observed disturbance in sodium and potassium levels in unorganized lead acid battery workers. After use of ACF mask, there is an improvement in sodium (−6.99%) and potassium levels (11.69%).
Renal manifestations of acute lead poisoning in animal models and in humans are observed as glycosuria, aminoaciduria [13, 14], and phosphaturia, collectively representing the Fanconi syndrome [15, 16, 17]. A generalized aminoaciduria observed in unorganized lead acid battery workers is indicated in Figure 2. After use of ACF mask, there was no reduction in the level of aminoaciduria this may be due to high concentration of lead in blood.
Battery manufacturing workers had severe complaints of lead toxicity like nausea, loss of appetite, constipation, intermittent abdominal pain, diarrhea, and joint pain. The acute and chronic symptoms of lead toxicity observed in battery manufacturing workers before using activated carbon mask were depicted in Figure 2. However, after using 2 months of activated carbon fabric mask, there was no significant decrease in acute and clinical symptoms, which may be due to high concentration of lead in the blood, soft tissues, and bones. The early recognition of these symptoms of lead exposure can minimize the toxic effects by using ACF mask. The acute and chronic clinical symptoms are reduced by decreasing the blood lead level.
This study is done for short duration as per study protocol. For long duration, we may get better results. We found use of ACF mask reduces blood lead and lessens renal toxicity. If the lead acid battery workers use such masks during working hours for long duration, definitely risk of lead toxicity is reduced.
In this study, activated carbon fabric mask was chosen because it is three-layered whereas other masks are two-layered. So ACF masks are better to protect from lead toxicity.
This is part of our study where we have studied effects of ACF on liver function, oxidative stress, and also on heme biosynthesis in occupational lead exposure which was published recently earlier. In our study protocol, neurological and cardiovascular studies are not included but in future we can plan.
In this study, Urine lead we could not do. Because blood lead level measured by usin lead analyzer Leadcare II machine. Which gives blood lead value within three minutes which was easy and most reliable. We did other biochemical parameters like urinary delta aminolevulinic acid. Urinary porphobilinogen these are biochemical markers for lead exposure. But our work on occupational lead exposure effect on heme biosynthesis and hematological properties published (Patil JA 2019 IJCB) hence not included.
Lead exposure is recognized as a risk factor for many of the diseases in battery manufacturing workers. This situation is worse in developing countries. Programs for the prevention and treatment of lead toxicity are poorly established [46]. Lead is environmental and occupational nephrotoxic cause of kidney disease, the present study aimed to reduce the renal toxicity.
Lead toxicity results in nephritis, which may be in association with hypertension. The symptoms of lead intoxication are variable. The assessment of workers with renal disease is important to identify the factors that may be leading to lead nephropathy. These factors are indications and risks of present or past lead exposure. Assessment of environmental and industrial lead is recommended to incorporate into programs for the prevention of chronic renal failure in countries where occupational lead exposure and toxicity still remain unchecked and burden of renal failure is increasing [46].
Therefore, it is essential to reduce the lead exposure by using special mask, apron, goggles, and even shifting the place of workers from high lead exposure to low lead exposure. In addition, unorganized lead acid workers need to be educated about the lead sources, hazards and that may cause lead toxicity. This is very important step in preventing further exposure to lead and its adverse health effects [5, 47]. Further prevention measures that involve the early detection and treatment of lead toxicity need to be started and also the targeted screening of workers with high risk of occupational lead exposure as this is likely to help early diagnosis and treatment of persons with lead nephropathy [47].
We acknowledge Late Dr. Arun Jalindar Patil, Deputy Director of Research, Executive Editor of JKIMSU, and Professor Dept of Biochemistry, who guided this project intellectually. We regret his absence.
We acknowledge our esteemed institute Krishna Institute of Medical Sciences Deemed to be University, Krishna Vishwa Vidyapeeth “deemed to be university”, for funding this project.
We are also thankful to the workers of unorganized battery industry who participated in this project.
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Written By
Jyotsna Patil, Arun Patil and Mandakini Kshirsagar
Submitted: 30 June 2023Reviewed: 21 November 2023Published: 09 January 2024
Open access peer-reviewed chapter
