Analysis of the Ecological Toxicity of Pesticides, Heavy Metals and Microbial Pollution and their combined effects, in the Running Surface Water of Jhal Magsi, Pakistan

doi:10.21203/rs.3.rs-4800354/v1

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Analysis of the Ecological Toxicity of Pesticides, Heavy Metals and Microbial Pollution and their combined effects, in the Running Surface Water of Jhal Magsi, Pakistan

https://doi.org/10.21203/rs.3.rs-4800354/v1

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Monitoring water quality is highly important for ensuring clean drinking water and protection of aquatic environments. The aim of the current study was to estimate the quality of running water sources from the district of Jhal Magsi. This study focused on evaluating the effects of low biodegradability, the accumulation of heavy metals and organics, and the presence of these compounds on water quality. The concentrations of herbicides (DDT (2,4 D) and Cyanazine) and insecticides (chlorpyrifos, Aldrin and Dieldrin) were determined via gas chromatography, and the concentrations of heavy metals (Pb and Cd) and trace elements (Zn and Mn) were detected via an atomic absorption spectrophotometer. Identification of E. coli and P. aeruginosa was performed by Gram staining and biochemical tests. DDT (2,4 D), Cyanazine, Chlorpyrifos, and Aldrin & Dieldrin were detected at 0.9 ± 0.058, 0.08 ± 0.006, 1.01 ± 0.577 and 1.1 ± 0.577 mg/L, respectively; these values are higher than the WHO safety limits. The heavy metals Pb and Cd were detected in all the samples, and the highest range for Pb was 2.03 ± 0.577 mg/L, while that for Cd was 0.48 ± 0.058 mg/L. The highest concentrations of the trace elements Zn and Mn were detected, and the WHO recommended safe limits were 0.305 ± 0.058 and 0.99 ± 0.058 mg/L, respectively. E. coli and P. aeruginosa were identified with percentages of 51.51 and 48.48%, respectively. The presence of higher concentrations of pesticides and heavy metals and their synergistic effect indicate a risk for both human health and aquatic organisms. Therefore, continuous monitoring of heavy metals and pesticides is necessary in the water reservoirs of Jhal Magsi to ensure drink safety. Bacterial assessment can aid in accepting bioremediation processes in water.

Food Science & Technology

Synergistic toxicity

Environmental monitoring

Agnostic relationship

Gas chromatography

Atomic absorption spectroscopy

Surface water, like lakes and rivers, is used for multiple purposes, supplying water for agricultural irrigation, drinking water, energy production and fisheries. However, these important resources are endangered by fast-rising populations, domestic and agricultural pollution, and developing technologies (Akanwa, 2021). The water pole is polluted by agricultural, sewage, and industrial waste. Metals and pesticides contaminate surface water via runoff waste. Environmental pollutants enter groundwater when rainwater parasites dissolve pollutants in the soil. Recurrent principles such as wineries often use trace elements, such as Cu or Zn. Heavy metals and pesticides accumulate in agricultural soils and finally spread to surface and ground water after rainfall (Madhav et al., 2020). Heavy metals can enter natural areas, but maximum contaminants originate from excess untreated wastewater discharge and spills, soil, and the dumping of human activities. Because of their persistent and nondegradable nature, pollution in water environments is an important problem that affects the environment, human health, and animals (Saravanan et al., 2022). Predicting the transportation of pollutants from soils to aquatic environments is important for assessing the risk and toxicity level of pollutants. The president's use of organochlorine pesticides was an important health fear. They can easily enter cell membranes because of their lipophilic nature. Anthropogenic activities release heavy metals into the environment because of the possible risk of contrary health impacts on both nontarget wildlife and humans (Davis, 2019; Kishor et al., 2021).

The toxicity of a component can change its effect on other components, causing unexpected health effects. This has posed an important challenge for ecotoxicology and environmental health in recent decades. Many studies in the literature have evaluated how different chemical mixtures interact and combined (synergistic or antagonistic, additive) toxicity occurs. In addition, numerous studies have been conducted to evaluate the acute toxicity of multicomponent chemical mixtures (Martin et al., 2021). Antagonistic or synergistic relationships occur when the toxicity of a mixture is less or greater than the sum of the toxicities of its components, respectively. In addition to these conflicting results, regulatory authorities typically impose regulations that treat acceptable concentrations of pollutants as independent, even when they are present in mixtures (Chen et al., 2015).

When water holds fecal material from industry, cattle farms, or hospitals, it contains loaded bacteria. Escherichia coli are known as a common indicator of fecal contamination. A bacteriological investigation of water contaminated by sewage revealed that sewage is unsafe for recreational or drinking purposes (some et al., 2021). Microbes can help to clean wastewater. They feed on the organic matter present in sewage and use nutrients for growth, which in turn enhances the cleaning process of wastewater. This action progresses the quality of water and its self-cleansing ability. Additionally, it helps to decrease the biochemical oxygen demand (BOD) and removes unpleasant odors (Brix, 2020).

For bioremediation, environmental conditions must be suitable for the growth and activity of these microorganisms. In nonideal conditions, adjustments are made to the environmental factors to increase the favorability of the growth of these microorganisms and therefore accelerate the degradation procedure. It is important to note that the microorganisms used in bioremediation are present in the environment and can stop any damage to the environment (Pal et al., 2020). In this study, we provide an overview of the pollution status of two major pollutants, pesticides and heavy metals, and how their synergetic effect can affect the environment. Additionally, we explored the prevalence of pathogen contamination in freshwater bodies in the Jhal Magsi district.

A total of 75 water samples were collected from different points in the rainy water resources (natural spring, canals and river water) of the district of Jhal Magsi Balochistan, Pakistan.

Quality Assurance and Preservation of Samples

The 1000 mL water samples were divided into three groups for detection of different parameters. One hundred milliliters of each sample was taken for bacterial analysis in separate labeled bottles. The remaining 500 mL of pesticide was removed, the sampling bottles were cleaned once with 30 mL of acetone, and 400 mL of sample was preserved by adding 5 mL of HNO₃ for heavy metal analysis. The samples were brought to the laboratory for analysis of bacteria within 24 hours.

Pesticide analysis

Water samples were extracted using a separatory funnel with 30 mL of dichloromethane solvent. The funnel was shaken for 5 minutes with periodic venting. After 15 minutes, the organic layer was separated from the water phase. The dichloromethane extracts were combined in a 250 mL Erlenmeyer flask. This extraction procedure was repeated three times, and the extracts were combined in the flask. The flask was then rotated at 200 rpm for one hour using a rotary evaporator. Interference was removed and purified by using silica gel columns. The extracts were evaporated and reconstituted with 0.5 mL of hexane for analysis. Quality control blanks of 1 liter of double distilled water contained no pesticides. The column temperature was programmed from 80 to 310°C with specific time intervals. A straight injection was used, and the injector temperature was maintained at 225°C (Ciornea et al., 2016).

The quantitative analyses of the selected pesticides were performed by gas chromatography, as shown in Fig. 2.

Heavy metals

For heavy metals, the digestion of water samples was carried out through the Fong SS technique (Mustafa et al., 2017).

Preparation of standard reagents

Standard solutions for each metal (Pb, Cd, Zn, and Mn) were prepared at different concentrations up to 1000 ppm (Fisher Chemicals, U.K.) by the formula given below. For the analytical determination, ultrapure compounds were used. Deionized water was used for distillation, after which the mixture was stored in glass containers (Asagba et al., 2007).

$$\:\text{V}\text{s}\left(\text{m}\text{g}\:\text{L}\right)=\frac{\text{C}\text{w}\:\times\:\text{V}\text{d}}{\text{C}\text{s}}$$

V_s = volume (mL) of primary standard (stock) added to the diluent to prepare the diluted working standard

C_w = Required concentration of working standard (mgL-1)

V_d = Desired final volume

C_s = Primary standard concentration of stock solution

Bacterial analysis

For bacterial isolation, water samples were analyzed. One hundred milliliters of each water sample (WHO, 2021) was added to 10 mL of broth (Luria Bertani). These falcon tubes were incubated at 37°C for 24 hours. The next day, the bacterial suspensions were cultivated on nutrient agar and then incubated at 37°C until the next day. Microscopic examination was performed by Gram staining, and biochemical identification was performed by different tests for the identification of E. coli and P. aeruginosa (Malik et al., 2023).

Statistical Data Analysis

The experiments were repeated at least in triplicate. All the pesticide and heavy metal data are presented as the average ± standard error according to the following formula

$$\:SE=\:\frac{\sigma\:}{\surd\:\text{n}}$$

SE = standard error

𝜎 = Standard deviation

√n = Numbers of Samples

The pesticides (herbicides and insecticides) removed from the running surface water of the district Jhal Magsi were DDT (2,4 D), Cyanazine, Chlorpyrifos and Aldrin & Dieldrin. The highest deducted ranges from the normal ranges of the WHO were 0.9 ± 0.058, 0.08 ± 0.006, 1.01 ± 0.577 and 1.1 ± 0.577 mg/L for DDT (2,4 D), Cyanazine, Chlorpyrifos and Aldrin & Dieldrin, respectively, as shown in Table 1.

Table 1

Pesticides in surface water samples from district Jhal Magsi
S. no	Sites	DDT (2,4 D)	Cyanazine	Chlorpyrifos	Aldrin & Dieldrin
		mg L-1
Types of Pesticides		Herbicide	Herbicide	Insecticide	Insecticide
Total no samples		54	52	49	31
WHO limits (MRL)		0.03	0.0006	0.03	0.00003
1	Peer chhatta	0.21 ± 0.058	0.02 ± 0.006	0.04 ± 0.006	0.87 ± 0.058
2	kotra	0.43 ± 0.058	0.04 ± 0.006	0.21 ± 0.058	0.01 ± 0.001
3	Gandawah	0.04 ± 0.006	0.06 ± 0.006	0.45 ± 0.058	0.76 ± 0.058
4	Gajan	0.01 ± 0.001	0.05 ± 0.006	0.09 ± 0.006	0.21 ± 0.058
5	Shoran	0.81 ± 0.058	0.04 ± 0.006	0.39 ± 0.058	0.09 ± 0.006
6	Mir Pur	0.9 ± 0.058	0.02 ± 0.006	0.68 ± 0.058	0.93 ± 0.006
7	Patri	0.31 ± 0.058	0.01 ± 0.001	0.91 ± 0.006	0.34v0.058
8	Kot Magsi	0.62 ± 0.006	0.03 ± 0.006	0.8 ± 0.058	0.72 ± 0.058
9	Mirpur jamot	0.51 ± 0.058	0.05 ± 0.006	0.04 ± 0.006	0.21 ± 0.058
10	sukleji	0.33 ± 0.058	0.08 ± 0.006	0.71 ± 0.058	0.4 ± 0.058
11	Gahela	0.21 ± 0.058	0.06 ± 0.006	0.04 ± 0.006	0.52 ± 0.058
12	Farom	0.03 ± 0.006	0.04 ± 0.006	0.31 ± 0.058	0.43 ± 0.058
13	Fathe Pur	0.53 ± 0.058	0.03 ± 0.006	1.01 ± 0.577	0.62 ± 0.058
14	Khari	0.49 ± 0.058	0.06 ± 0.006	0.68 ± 0.058	1.1 ± 0.577
15	River Mula	0.31 ± 0.058	0.04 ± 0.006	0.45 ± 0.058	0.67 ± 0.058
*All selected pesticides were measured (average ± standard error (SE)) in mile grams per litter (mg/L) in all the water samples. The maximum residue limits (MRLs) of various pesticides in drinking water were determined (WHO, 2017)

The heavy metals Pb, Cd, Zn and Mn were analyzed from the surface running water of the district of Jhal Magsi. Pb and Cd were detected in all the district samples, and the highest ranges of Pb, Cd, Zn and Mn were 2.03 ± 0.577 mg L-1, 0.48 ± 0.058 mg L-1, 0.305 ± 0.058 mg L-1 and 0.99 ± 0.058 mg L-1, respectively (table. 2).

Table 2

Heavy metals in the surface water of district Jhal Magsi
S .no	Name of area	Pb	Cd	Zn	Mn
		mg L-1
Type of metals		Heavy metal	Heavy metal	Trace metal	Trace metal
Total present metals		67	64	66	59
EPA/WHO (MRL)		< 0.05/0.01	0.01/0.003	5/3	< 0.5/0.5
1	Peer chhatta	1.72 ± 0.577	0.09 ± 0.006	0.088 ± 0.006	0.16 ± 0.058
2	kotra	1.64 ± 0.577	0.105 ± 0.058	0.083 ± 0.006	0.101 ± 0.058
3	Gandawah	1.97 ± 0.577	0.104 ± 0.058	0.061 ± 0.006	0.11 ± 0.058
4	Gajan	2.03 ± 0.577	0.11 ± 0.058	0.058 ± 0.006	0.104 ± 0.058
5	Shoran	1.98 ± 0.577	0.21 ± 0.058	0.088 ± 0.010	0.27 ± 0.058
6	Mir Pur	1.87 ± 0.577	0.34 ± 0.058	0.101 ± 0.058	0.12 ± 0.058
7	Patri	1.988 ± 0.577	0.1 ± 0.001	0.305 ± 0.058	0.99 ± 0.058
8	Kot Magsi	1.78 ± 0.577	0.48 ± 0.058	0.099 ± 0.006	0.44 ± 0.064
9	Mirpur jamot	1.88 ± 0.577	0.101 ± 0.058	0.097 ± 0.001	0.34 ± 0.058
10	sukleji	1.96 ± 0.577	0.104 ± 0.058	0.088 ± 0.006	0.14 ± 0.058
11	Gahela	1.99 ± 0.577	0.102 ± 0.058	0.078 ± 0.006	0.13 ± 0.058
12	Farom	1.98 ± 0.577	0.103 ± 0.058	0.098 ± 0.001	0.87 ± 0.058
13	Fathe Pur	1.88 ± 0.577	0.11 ± 0.058	0.102 ± 0.058	0.34 ± 0.058
14	Khari	1.78 ± 0.577	0.103 ± 0.058	0.205 ± 0.058	0.37 ± 0.058
15	River Mula	1.86 ± 0.577	0.099 ± 0.006	0.169 ± 0.058	0.28 ± 0.058
*Pb (lead), Cd (cadmium), Zn (zinc) and Mn (manganese). All the heavy metal concentrations were reported as average ± standard error (SE)) in milligrams per litter (mg/L) in all the water samples. The maximum residue limits (MRLs) of various heavy metals in drinking water were calculated (WHO, 2021)

From the samples of all the selected sites, E. coli and P. aeruginosa were identified. There were 51.51 and 48.48% E. coli and P. aeruginosa, respectively. At the Gandawah sample site, both bacteria were identified at the same ratio of 4/5 (table. 3).

Table 3

Total number and percentage of bacteria
S .no	Name of area	E. coli	P. aeruginosa
1	Peer chhatta	3/5	3/5
2	kotra	2/5	2/5
3	Gandawah	4/5	4/5
4	Gajan	3/5	3/5
5	Shoran	1/5	1/5
6	Mir Pur	1/5	1/5
7	Patri	2/5	2/5
8	Kot Magsi	3/5	3/5
9	Mirpur jamot	3/5	3/5
10	sukleji	1/5	1/5
11	Gahela	1/5	1/5
12	Farom	2/5	2/5
13	Fathe Pur	2/5	2/5
14	Khari	1/5	1/5
15	River Mula	3/5	3/5
Total no of Bacteria		34/75	32/75
Percentage of Bacteria		51.51%	48.48%

*From every site, five (5) water samples were collected for bacterial analysis, and a total of 75 water samples were used for bacterial detection.

The data were analyzed according to the central limit theorem. Therefore, Pearson’s correlation was used. The correlations are shown in Table 4. According to the results of correlation analysis, Pb has shown an inverse relationship with Cd, DDT (2,4 D), Aldrin and Dieldrin, E. coli, and P. aeruginosa; Cd has a correlation with chlorpyrifos, DDT (2,4 D), and Aldrin and Dieldrin; Zn has a strong correlation with Mn and chlorpyrifos; DDT (2,4 D) has a correlation with chlorpyrifos; and chlorpyrifos has a correlation with alrin and Dieldrin (p < 0.01).

Table 4

Pearson correlation chart for heavy metals, pesticides and bacteria
	Pb	Cd	Zn	Mn	DDT (2,4 D)	Cyanazine	Chlorpyrifos	Aldrin & Dieldrin	E. coli	P. aeruginosa
Pb	1
Cd	-0.241	1
Zn	0.011	-0.102	1
Mn	0.188	0.016	0.677**	1
DDT (2,4 D)	-0.307	0.493*	0.107	-0.178	1
Cyanazine	0.256	-0.321	-0.383	-0.414	-0.304	1
Chlorpyrifos	0.026	0.371*	0.503**	0.374	0.400*	-0.235	1
Aldrin & Dieldrin	-0.243	0.335*	0.155	-0.015	0.084	-0.090	0.333*	1
E. coli	-0.091	0.041	-0.187	0.019	-0.474	-0.132	-0.237	0.025	1
P. aeruginosa	-0.091	0.041	-0.187	0.019	-0.474	-0.132	-0.237	0.025	1.000	1

*Pb (lead), Cd (cadmium), Zn (zinc) and Mn (manganese). **The Pearson correlation is significant at the 0.01 level (2-tailed). *Correlation is significant at the 0.05 level (2-tailed), and –ive indicates an inverse relationship.

Table 5

Relationships between Exposure to Heavy Plants, Pesticides and Their Combinations
S.no	Metals	Pesticides	Effects	References
1	Cd,	Chlorpyrifos	Cause damage and Oxidative stress to the mitochondria in neuronal cells.	Xu et al. (2017)
2	Zn	Chlorpyrifos	Cause cellular stress responses and reproduction problems.	García-Gómez et al. (2019)
3	Ni, Mn, Pb, Cd, Zn	Different pesticides, PCB and chemical	Combine effect the heavy metals residual attractive forces and Unbalanced forces (adsorption mechanism).	Yang et al. (2020)
4	Cd, Pb	Chlorpyrifos	It cause combined toxicity to daphnia magna was preservative	Yongmeng et al. (2021)
5	Cd	Diazi on	Seminiferous epithelium, cause loss of spermatogenic element, lacking maturation of germs cells and disorganization	Singh et al. (2017)
6	Ni, Mn	Chlorpyrifos	Change in molecular fingerprints and giving rise to a complex transcriptional profile in mixture	Vandenbrouck et al. (2009); Boatti et al. (2012)
	Cd	Diazinon	progressive damage in epithelial cells and decrease in spermatogenic	Adamkovicova et al. (2014)
7	Cd	Chlorpyrifos	Because headaches, seizures, blurred vision, salivation, irritate the stomach, coma, and result in diarrhea and vomiting.	Nandi et al. (2022); Mohod and Dhote (2013)
8	Cd	2,4- D, DDT	Tremors or shakiness, vomiting, and seizures. DDT is carcinogen and exposure can affect the liver and reproduction.	Strong et al. (2015); Mohod and Dhote (2013)
9	Pb, Cd	Chlorpyrifos	Effect nervous system, blurred vision, ranging from headaches, seizures, coma, and salivation depending on the amount and length of exposure.	Nandi et al. (2022); Mohod and Dhote (2013)

*Pb (lead), Cd (cadmium), Zn (zinc), Ni (nickel), Mn (manganese) and PCB (polychlorinated biphenyl). The A column shows the combined effect of heavy metals and pesticides.

The investigations were carried out on the running surface water of the district of Jhal Magsi. Different pesticides and fertilizers used for the proper growth of plants. Exposure to heavy metals and pesticides has caused a high number of cancer cases. A current risk assessment, which also included data from animals, estimated that exposure to dioxin-like and dioxin compounds (such as PCBs) could result in cancer up to 10^− 4 per year.

Burns et al. (2021) researched pesticide sources and their effects on humans. In the present study, many pesticides were identified via chromatographic experiments with a standard solution. The same technique was utilized by Wang et al. (2007) and Nguyen et al. (2019) for the quantification and determination of pesticides in surface water through gas chromatography–mass spectrometry. In the current research conducted by Júnior and Re-Poppi (2007) and Cortada et al. (2009), various pesticides, including Aldrin and Dieldrin, were detected in water samples. These pesticides were found in river, surface, and tap water, with higher concentrations observed in surface water samples. Tiwari and Guha (2013) evaluated chlorpyrifos, an organophosphorus pesticide, in an aqueous environment. Arain et al. (2018) studied the chlorpyrifos concentration in the surface and ground water of Okra. These results are consistent with the findings of the present study. Chlorpyrifos was detected in all the water samples from the district.

In the present study, cyanazine was detected in water samples; Schraer et al. (2000) also detected cyanazine in surface water. Cyanazine has acute toxicity and is moderately toxic to mammals. Furthermore, the herbicide DDT was also detected in the water of the district Jhall Magsi (Strong et al., 2015; Burgos-Aceves et al., 2021). Contact with DDT can cause contrary effects on reproduction and the liver. High doses of DDT can cause seizures, vomiting and tremors in humans.

In the present study, Cd and Pb were detected at relatively high concentrations, and Zn and Mn were detected in the majority of the samples within the limits of the WHO. These results are consistent with those of Hange and Awofolu (2017), who assessed the heavy metal concentrations in river surface water collected from the Jhelum River at Muzaffargarh. The concentrations of Ni, Pb, Cd, and Cr were found to be higher than the WHO limits in drinking water, and the concentrations of metals such as Mn and Zn were lower than the WHO standards for water (Ajiwe et al., 2018). Mohiuddin et al. (2016) studied sediment and water samples from the Buriganga River of Bangladesh and reported that the Cd, Pd, Zn, Cu, and As concentrations exceeded the toxicity reference values. Mohod and Dhote (2013) reported that human sources can be categorized according to the main cause and route through which heavy metals are distributed to different sections of the environment and through which the transport media is mainly water. In the present study, the sources of running water were the Mula River, the Sukhlaji River, natural springs and canal water. These reservoirs are contaminated through natural and anthropogenic activities. Increases in socioeconomic activity due to increased population have been connected to rapid increases in heavy metals in the environment. Anthropological activities such as agriculture, ore processing, and mining contribute greatly to metal contamination. Domestic activities, such as laundry detergents, also contribute to heavy metal concentrations in water, which has opposite effects (Minhas et al., 2022). The main water reservoirs of the district Jhal Magsi follow the routes of mountain and agricultural fields, so weather plays a role in the contamination of water with metals and pesticides (Mustafa et al., 2017). According to Négrel et al. (2018), weathering is a natural process in which rocks, minerals, and soil are broken down through atmospheric exposure, living organisms and water. This process can release heavy metals through numerous natural events, such as decay comets, volcanic eruptions, erosion and heavy rainfall. Liu et al. (2020) reported higher concentrations of Pb and Cd than the WHO permissible limits in a study conducted on a river containing Swat water. Briffa et al. (2020) studied Hg, Cd, and Pb in the surface and ground water of Central East India, and their results also revealed higher concentrations of Pb and Cd in water samples. According to Ajiwe et al. (2018), heavy metals are carcinogenic, genotoxic, teratogenic and mutagenic.

Inamori and Fujimoto (2010) studied the safety of drinking water by measuring bacteria in 1 mL of test water. There were fewer than 100 bacterial colonies, and no total coliforms were detected. These results contrast with those of the present study in which 1 mL of water sample contained E. coli and P. aeruginosa. E. coli and P. aeruginosa were detected in water by Jawad et al. (2020) and Baghal et al. (2021) and in tap water (Moghadam et al., 2016).

The growth of bacteria is activated by an increase in the concentration of phosphorus and nitrogen, which are commonly found in pesticides. This increase often occurs due to contamination from sources such as industrial sewage, livestock sewage, and domestic sewage, which may contain heavy metals. Pathogenic bacteria that cause water-borne diseases can contaminate the human intestine and cause runoff into the environment through manure. These bacteria then undergo action and are ultimately released into water reservoirs, lakes, and rivers (Some et al., 2021).

Water cleaning involves the use of microbes as biological agents to eliminate or reduce the effects of environmental pollutants. Microbes are utilized mainly because of their rapid growth and ability to be manipulated easily; thus, they function as agents for bioremediation (Brix, 2020). The results of the present study will help future readers study the role and mechanisms of bioremediation.

The combined pollution of pesticides and heavy metals poses a serious risk to human living and the soil ecological environment. It has been reported that pesticides and heavy metals enter animal and human bodies via ingestion through food materials (Satarug et al., 2017), dermal contact and inhalation, smoke formation, fumes of chemicals, dust particles, numerous activities, mining, and battery manufacturing (Asgary et al., 2017). The combined pollution of Cu and chlorpyrifos had synergistic and antagonistic effects on different tests (Yongmeng et al., 2021). Cd exposure increases the susceptibility to microbial pesticides in a synergistic manner, and the use of microbial pesticides is an effective strategy for pest control in heavy metal-polluted areas (Zheng et al., 2023). Microorganisms can convert hazardous Hg²⁺ and Cr⁶⁺ ions into less toxic (Hg⁰⁺) and Cr³⁺ ions (Malik et al., 2023). Ni, Cd and chlorpyrifos have synergistic effects on the environment and can cause the conversion of Cd2 + and Ni²⁺. Lam et al. (2023) examined the mutual effect of 2,4-dichlorophenol laterally with Cu and Zn.

In the present study, pesticide, heavy metal and microbial assessments of water samples were performed and compared with the recommended limits of the WHO. DDT (2,4 D), Cyanazine, Chlorpyrifos, Aldrin and Dieldrin pesticides and Pb and Cd were found to be present at concentrations higher than the WHO standard limits. The predicted synergistic effects resulting from the co-occurrence of these pesticides and heavy metals represent an enhanced risk to humans, and additional bacterial assessments could aid in understanding the role of these metals by enhancing the cleaning and bioremediation of polluted water.

Author contributions

Conceptualization: S.M. and A.-U.-R.K.; Data curation: Y.M., S.M. and N.K.; Investigation: S.M., A.-U.-R.K., Y.M., N.K., A.S., M.A and S.; Methodology: S.M., Y.M. and S.; Supervision: A.-U.-R.K., N.K. and A.S.; Validation: S.M. and Y.M.; Writing original draft: S.M., Y.M. and A.-U.-R.K.; Writing - review & editing: N.K., A.S. and M.A

Competing interest: No competing interests exist.

Funding: This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Acknowledgments: None

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The authors declare no competing interests.

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Analysis of the Ecological Toxicity of Pesticides, Heavy Metals and Microbial Pollution and their combined effects, in the Running Surface Water of Jhal Magsi, Pakistan

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