The Effect of Water Treatment Models to Reduce Lead (Pb) Level on Freshwater Snail Filopaludina javanica
Diana Arfiati*, Nur Syahid, Zaki Anwari, Aminin Aminin, Kusriani Kusriani, Endang Yuli Herawati, Asthervina Widyastami Puspitasari
Faculty of Fisheries and Marine Science, University of Brawijaya, Indonesia Email Address : [email protected]
Abstract Lead is a kind of non-essential heavy metals included in the metal causing environmental pollution with persistent properties that might harm the consumers. This study aimed to determine the best method for reducing the lead level on the freshwater snail Filopaludina javanica using three various water treatment models. Soaking water treatment (6h, 12h, 18h, 24h), flowing water treatment (6h, 12h, 18h, 24h), and refreshing water treatment (6h, 12h, 24h).
The lead level assay in both samples used Atomic absorption spectroscopy (AAS), and the physical and chemical parameters were measured, such as temperature, pH, and Dissolved Oxygen (DO). The lowest value of lead content on freshwater’s soft body snail and water sample respectively were soaking water treatment at 6h (0.64 ± 0.02 mg L-1) and 24h (0.0045 ± 0.0015 mg L-1); flowing water treatment at 24h (0.04 ± 0.007 mg L-1) and 18h (0.0036 ± 0.0009 mg L-1) and; refreshing water treatment at 24h (0.150 ± 0.011) and 12h (0.007 ± 0.001), with control 0.072 ± 0.00 mg L-1 and 0.067 ± 0.00 mg L-1. Therefore, the most effective model to reduce the lead content was flowing water treatment within 24h in the freshwater soft body snail and 18h in the water sample.
Introduction
Heavy metals are classified as metallic elements with a high atomic weight and a density of at least 5 times greater than that of water (Tchounwou et al., 2012). The toxicity and bioaccumulation propensity of heavy metals in the environment become a severe threat to living organism health. The chemical or biological processes cannot remove heavy metals. Hence, they will be transformed into less harmful species such as Jabon (Anthocephalus cadamba Roxb), which potential for remediating Lead (Pb) (Ayangbenro & Babalola, 2017; Setyaningsih et al., 2018). The majorities of heavy metals with low toxic levels are competent in entering the food chain, which is accumulated and causes adverse effects in the living organism. The
harmful effect of heavy metals depends on each metal's amount on the body, the absorbent dose, the route, and the exposure time (Mani & Kumar, 2014).
Lead (Pb) is a non-essential heavy metal that is ubiquitous and abundant in the Earth's crust since prehistoric time (Tong et al., 2000).
It has been distributed and mobilized in the environment. Pb naturally exists in the rocks, soil, water, and air. Currently, In the entire world, the high quantity levels of lead found in the environment through the entire world come from human activities such as burning fossil, fuels, mining, and manufacturing (Tiwari et al., 2013). Lead exposure remains a serious problem in many developing and industrializing countries, mainly in the water environment. The toxicity of lead to human KEYWORDS
Environmental pollution;
Filopaludina javanica;
Lead;
water treatment.
causes anorexia, chronic nephropathy, neurons dysfunction, hypertension, hyperactivity, insomnia, learning deficiency, reduced fertility, renal system damage, a risk factor for Alzheimer's disease, shortened attention span, to the plant's effects on photosynthesis and growth, causes chlorosis, inhibits enzyme activities and seed germination, and becomes sources of oxidative stress, for microorganisms denatures nucleic acid and protein, inhibits and transcribes enzymes activities (Nagajyoti et al., 2010; Fashola et al., 2016; Mupa et al., 2013; Wuana et al., 2011).
Waung river is one of the rivers located in the Lamongan district, Indonesia. The Waung river condition was contaminated by household wastes such as laundry activities.
The protection for water environments, especially freshwater sources like rivers and streams, is necessary to prevent environmental deterioration and reduce living organism biodiversity (Edokpayi et al. 2016).
Natural processes happening on land caused by the fresh flow water flow into the sea through the river, and the influence of a variety of human activities on land involving sedimentation and pollution forms (National Strategy and Action Plan, 2015). Water analysis is commonly used to quantify the accumulation of heavy metals in water pollution.
Filopaludina javanica or known as Bellamya javanica von Dem Busch, 1844, is one of the freshwater snails which is originally from Indonesia. In nature, a freshwater snail can obtain the food by filter-feeding, which can easily accumulate pollutants, including heavy metals in its tissues. Besides its potential as freshwater heavy metal accumulators, snails are also used as animal feed ingredients such as duck feed and local food in the
on consumers. For that reason, efforts should be made to reduce the heavy metal content in the filter feeder organism by using effective methods such as the water treatment method.
However, this study's objective was to determine the most effective method for the lead exposure reduction exposure using water treatment on the freshwater snail Filopaludina javanica as an animal model.
Materials and Methods Sample Preparation
Waung river is located in the Glagah sub- district, Lamongan district, Indonesia (7°03'42.5" S 112°29'35.1" E), which is 4 m wide, depth 1 to 2 m, and 7 km long to Bengawan Solo river. The chosen sampling area was divided into 3 stations based on the feasibility of finding freshwater snails.
Freshwater snails (Filopaludina javanica) were collected in July 2017 from the Waung river with a depth of 20 cm below the surface. The freshwater snails used had 25 mm to 27 mm were kept in a 5-L plastic tank with 5 cm soaked using river water then were taken to the Fisheries and Marine Science Laboratory of University of Brawijaya immediately. The soft body of 15 freshwater snails and river water were taken directly and were tested on the Pb level as a control. In this study, the three alternative methods were used for reducing the Pb levels in the freshwater snails, including soaking water treatment, flowing water treatment, and refreshing water treatment.
During the acclimatization, the freshwater snails were neither fed nor aerated for about 24 hours.
Water Treatment Analysis Methods Soaking Water Treatment Method
This part was conducted in 24 tanks prepared, separated into 4 groups (A group, B
(2014) compared to the control. In this method, each tank with a diameter of 58 cm was filled with 20 cm deep of pure water and 12 freshwater snails and was acclimatized approximately 24 hours with aeration.
Different time points soaked each sample group. Those were 6 hours (A group); 12 hours (B group); 18 hours (C group); and 24 hours (D group). Following that, the Pb levels were checked by Atomic absorption spectroscopy (AAS) (Shimadzu 6800AA). Every passing of each time point, 5 samples of the freshwater snails were taken and sacrificed from each tank by separating the freshwater snails' body and shell. The soft body of freshwater snails and the water on the tank were used to detect Pb levels. Besides the Pb levels, the physicochemical parameters were tested following the group. The physicochemical parameters included temperature (Omron), dissolved oxygen (Lutron DO-5510), and pH (Test 30).
Flowing water Treatment
In this part, 24 tanks with a diameter of 58 cm were prepared and were filled with as much as 20 cm deep of pure water, and 12 freshwater snails were acclimatized for 24 hours. Each group was performed in six replication following Federer (2014). In this method, each tank was watered for 24 hours with aeration, and the samples were collected at different time points following each group to check the Pb levels on the soft body of the freshwater snail and in the water sample in every treatment by using Atomic absorption spectroscopy (AAS) (Shimadzu 6800AA). The physicochemical parameters were also analyzed from each group, including A group watered for 6 hours; B group watered for 12 hours; C group watered for 18 hours, and the D group watered for 24 hours and the
results were compared to that of the control (0 hours). The flow velocity in every group was about 0.4-0.5 m s-1.
Refreshing Water Treatment
The principle in this method is similar to the soaking water treatment method. The freshwater snails were soaked at different time points, followed by 24 hours. The samples were separated into 4 groups, and each group was performed into six replication. Control group was conducted at a 0-hour time point; A group was conducted at 6 hours time point with four-time water changes; B group was conducted at 12 hours time point, and the water was changed twice, and C group was conducted at 24 hours time point with no water change. The Pb levels and physicochemical parameters were tested in each treatment group. The Pb levels in the soft body of the freshwater snails and the water of each group were tested by Atomic absorption spectroscopy (AAS) (Shimadzu 6800AA).
Statistical Analysis
The statistical analysis was determined by a one-way analysis of variance (ANOVA) in SigmaPlot ver 12.0 and SPSS ver.16 software followed by Tukey's test. Data are expressed as a mean ± standard deviation, which statistically significant differences required that p < 0.05.
Results and Discussion
Bio-availability of heavy metals in aquatic ecosystems are strongly dependent on physicochemical parameters such as temperature, pH, and dissolved oxygen; the Physico-chemical parameters results of different alternative methods are shown in Table 1.
Table 1. Sample Measured of Physico-Chemical Parameters
No. Model Time Point
(h)
Temperature (°C)
pH DO
(mg/L) 1. Water Sampling from Waung
River (control) 0 31.1 6.9 3.9
2. Soaking Water Treatment 6h 27.1 7.3 7.7
12h 28.6 7.9 8.2
18h 29.2 7.6 8.5
24h 28.6 7.4 8.1
3. Flowing Water Treatment 6h 27.1 7.3 5.5
12h 28.6 7.3 5.1
18h 29.9 7.3 4.7
24h 28.6 7.4 5.1
4. Refreshing Water Treatment 6h 26.4 8.1 8.0
12h 26.4 8.1 7.9
24h 26.2 8.1 8.0
The value indicated the means of samples (n=6).
Physical and chemical test parameters of water quality in this study included were temperature, pH, and dissolved oxygen.
Seuffert and Martin (2017) reported that the wide temperature of freshwater snail P.
canaliculata range from 15 to 35°C, and the optimum temperature is 25°C. The range of three alternative methods of temperature was from 26.3 to 28.5°C. This indicates that these temperatures are suitable for freshwater snail life. The results show that each water treatment model has different physicochemical parameters (Figure 1). The temperature decreases compared to the control in every method. This phenomenon can be caused by the fact that the Waung river is an outdoor place with direct sun exposure, while the laboratory condition is protected from the sunlight. pH level showed an increased value compared to the control. pH is a substantially necessary factor that affects
was 7.3-8.1. In this condition, the pH is still appropriate for the freshwater snail life cycle.
Freshwater snails are proved to be completely sensitive to acidification environments (Raddum et al., 1988). Freshwater snails are tolerant to low pH as long as there is a sufficiency in Ca2+ (Okland, 1983).
Dissolved oxygen (DO) shows increasing levels except in the b method; the average DO of the b method is 5.1 mg L-1. This is possible because the b method used a flowing water treatment, which means the oxygen content always changed from new water, whereas the DO level in the original water is about 3.9 mg L-1. Conversely, a and c methods show a higher level of dissolved oxygen (DO) content at 8.1 and 8.0 mg L-1, respectively, than the control group. The fact that the water does not flow can contribute to the depositing oxygen on the tanks. The standards of DO, pH, and temperature for freshwater snail life are 3-7
Figure 1. The level of physicochemical parameters. a) Soaking water treatment; b) Flowing water treatment; c) Refreshing water treatment. The statistical differences of each group were assay with ± SD (n=6).
Table 2. The Lead Level on Water and Soft Snail Body Samples
Model Time
Points
Pb Level
Water Sample Soft Body Sample
Soaking Water Treatment
0h 0.067c ± 0.000 0.072a ± 0.007
6h 0.006b ± 0.0008 0.064a ± 0.019
12h 0.006ab ± 0.0012 0.111b ± 0.024 18h 0.005ab ± 0.0011 0.157c ± 0.027
24h 0.005a ±0.0015 0.185c ± 0.026
Flowing Water Treatment
0h 0.067c ± 0.000 0,072c ± 0.007
6h 0.006b ± 0.001 0.076c ± 0.011
12h 0.005b ± 0.001 0.056b ± 0.011
18h 0.004a ± 0.001 0.046ab ± 0.005
24h 0.005b ± 0.001 0.040a ± 0.007
b)
Physico-Chemical Parameters
Temperature (oC) pH DO (mg/L)
Level
0 5 10 15 20 25 30
35 Control
6h 12h 18h 24h
Physico-Chemical Parameters
Temperature (oC) pH DO (mg/L)
Level
0 5 10 15 20 25 30
35 Control
6 h 12h 18h 24h
c)
Physico-Chemical Parameters Temperature (oC) pH DO (mg/L)
Level
0 5 10 15 20 25 30
35 Control
6h 12h 24h
Model Time Points
Pb Level
Water Sample Soft Body Sample
Refreshing Water Treatment
0h 0.067c ± 0.000 0.072a ± 0.007
6h 0.004bc ± 0.000 0.140bc ± 0.010
12h 0.007b ± 0.001 0.115b ± 0.010
24h 0.003a ± 0.001 0.150c ± 0.011
The value indicated the means of samples (n=6). All data were assayed with standard deviation (SD) with p < 0.05
The lead level on the water and soft body samples showed different results in Table 2. All of the water samples' results in all water treatment models showed a significant difference compared to the control. Contrast to the soft snail body samples results, which show the different pattern in a different water treatment model. The lead content on the water sample with soaking water treatment showing the lowest value was 24 hours after treatment, and the highest value was found in the 6 hours after treatment. The Pb level in the flowing water treatment showed the lowest value at 18 hours and the highest value at 6 hours. Meanwhile, in the refreshing water treatment, the lowest value was found at the 24-hour time point, and the highest value was at the 12-hour time point. Furthermore, the lead contents of the water samples of all models were lower compared to the control (Figure 2A). In contrast, the soft snail body samples result showed that not all lead levels of all water treatment models decreased (Figure 2B). The lead content on the soft body of the soaking water treatment's freshwater snails was highest at the 24-hour time point and lowest at the 6-hour time point. On the other hand, in the flowing water treatment, the highest value of lead was found at the 6-hour time point, and the lowest value of lead was found at the 24-hour time point. Whereas, in the refreshing water treatment, the highest value of lead was indicated at the 24-hour time point, and the lowest value of lead was shown at the 18-hour time point.
Figure 2. a) The Pb level (mg L-1) on the freshwater soft body; b) The Pb level (mg L-1) on the water sample compared to each treatment in every water treatment method. The value was carried out by six times of repetition.
The treatment symbol means a. Soaking water treatment; b. Flowing water treatment; c. Refreshing water
This study proves that the water sample in all water treatment models showed similar pattern results (Figure 2A). This phenomenon is evident that there has been a reduction of Pb's heavy metal content in the water. It indicates a correlation between the remaining Pb content in the water with the metal uptake process by freshwater snail Filopaludina javanica. Besides that, the physicochemical parameters influence the lead content in the water, but there is no direct relationship between the accumulation value of a lead in the freshwater soft body. Those explanations are supported by Di Toro et al. (2001), and Santore et al. (2001), The bioavailability and toxicity of metals to aquatic organisms depend on the water chemistry factors such as hardness, salinity, specific ion levels, pH, alkalinity, complexing agents and dissolved organic carbon (DOC). Afterward, the concentration of the same metal in different chemistry waters might result in differential toxicity. The freshwater soft body result showed that in the soaking water treatment model and refreshing water treatment model occurred an increasing significantly time by time in lead content, but different from the flowing water treatment model. The result showed decreased lead content in the flowing water treatment model significantly in time by time, especially compared to control (Figure 2B). This happens because of the freshwater snail ability to absorb the lead through metal uptake processes. Likewise, internal metal bioavailability is important because toxicity is not related to the total accumulated metal concentration of internal metabolically available metal (Rainbow, 2007). In a freshwater ecosystem, Filopaludina javanica is one of the gastropods that can be a bioindicator of heavy metals. Its character is a filter feeder accumulating pollutants, including heavy metals in its tissues, sub-hazards for the consumers,
especially humans. In aquatic organisms, heavy metals uptake from the water source through a cellular basis in different ways (Rainbow, 2002). The response of different species also plays an important role in metal bioaccumulation in tissues and organs (Tessier et al., 1994). Oros et al. (2010) explained in their experiment showed gastropods and bivalve mollusks have a different ability to accumulate toxic metals.
Lead, along with other metals, can substitute for calcium ions and incorporate into calcium carbonate crystals in the shell composition. It is suspected that it happens by its mechanism of any metal embedded in shell structure was uptaken from the environment and metabolized by the organism. Metal accumulated in higher concentrations in tissues directly exposed (gills, skin) or involved in detoxification (liver, kidney) and less in muscle. In bivalve mollusks (invertebrate) have shown the importance of factors such as food quantity, composition, and concentration of metals in food. The accumulated metals on its mechanism through sediment particles, organic compounds (such as bacterial extracellular polymers and fulvic acids), or the appearance of benthic microalgae significantly enhance this process (Reinfelder, 1998). Furthermore, the different patterns in the soft body result might be caused by environmental influences.
In the soaking water treatment and refreshing water, treatment models experienced more stress than the flowing water treatment model. For the soaking water treatment model, there was no fresh water changing, this condition makes the water concentration, and for refreshing water treatment model, the water change continually affects freshwater snail occurred the environmental stress, thereby affecting the physiologically stressing and impairing the heavy metal defense responses of a freshwater snail
(Lefcort et al., 2015). The ecological and ecophysiological studies explain that mollusks react to environmental stress and pollution by modifying behavior and various life-history traits in a manner consistent with maintaining population fitness (Sibly and Calow, 1989;
Calow, 1991).
CONCLUSIONS
Flowing water treatment is the best model for reducing the lead level in the Filopaludina javanica soft tissue. The lead concentration on the soft body of freshwater snail decreases by 189% in 24 hours post- treatment compared to the control. This method effectively maintains the lead accumulation on the soft body of freshwater snail before consuming and diminishing the adverse effect. For the future, the histopathological and molecular assay will be needed to confirm the independent mechanism of a freshwater snail (Filopaludina javanica) in reducing the lead concentration.
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