Analyzing the impact of pesticides on the indus river: contamination levels in water, sediment, fish, and associated human health risks.

Pesticides are frequently used to protect crop yields and manage malaria vectors; however, their inadvertent transport into aquatic habitats poses a significant concern. Various anthropogenic activities influence the Indus River in Pakistan. This study aimed to assess the presence of eight pesticide residues at three different sites (Kalabagh, Kundian, and Chashma) in water, sediment, and the fish species (Labeo rohita) during both dry and wet seasons to measure the intensity of this pressure. Pesticide analysis was carried out using gas chromatography equipped with an electron capture detector. The results revealed the highest concentrations of pesticides during both dry and wet seasons at all sites, measuring 0.83 and 0.62 µg/l (water), 12.37 and 9.20 µg/g/dw (sediment), and 14.27 and 11.29 µg/g/ww (L. rohita), respectively. Overall, pesticide concentrations were higher in the dry season than in the wet season across all study sites. Based on detection frequency and concentration in both seasons at all sites, dominant pesticides included cypermethrin and carbofuran (in water), as well as endosulfan and cypermethrin (in sediment and fish tissue). Levels of endosulfan and cypermethrin exceeded standard limits. Moreover, principal component analysis (PCA) indicated no correlation among pesticides in fish tissue, sediment, and water. However, pesticides exhibited different behavior in different seasons. Furthermore, endosulfan and triazophos impose great human health risk, as indicated by the THQ value (> 1). The overall HI value was greater for site 1 in the dry season (8.378). The study concluded that the presence of agricultural pesticides in the Indus River poses a risk to aquatic life and has the potential to disrupt the entire food chain. This highlights the importance of sustainable practices for the study area and Pakistan overall agricultural and environmental sustainability. It is further recommended to strengthen regulations for reduced pesticide use and promote eco-friendly pest management.

Sublethal Concentrations of Cadmium and Lead: Effects on Hemato-Biochemical Parameters and Tissue Accumulation in Wallagu attu.

The exposure of fish to heavy metals can significantly impact physiological processes and potentially lead to adverse health effects. This study assesses the effects of exposure to Cd and Pb sublethal concentrations in water on Wallagu attu. A total of 48 fish with an average body weight of 145.5 ± 26 g were distributed among three groups (control, Cd-treated, and Pb-treated) within 60 L fiberglass tanks. They were exposed to 30% sublethal concentrations of Cd and Pb for durations of 1, 15, and 30 days. Following this exposure, an assessment was conducted on metal bioaccumulation and hemato-biochemical responses. Results revealed a significantly (P < 0.05) higher concentration of heavy metals in the fish tissues of metals exposed groups than in the control. The concentration of Cd and Pb increases in fish tissues (kidney > gills > intestine) with exposure time. In most cases, the Pb-exposed group exhibited significantly (P < 0.05) higher concentrations of Pb in different tissues than the Cd-treated group. With extended exposure time, the activities of CAT and SOD show a significant decrease in both Cd and Pb-treated groups. However, the reduction in activities was more pronounced in the Cd-exposed group. On 15 and 30 days, the levels of red blood cells (RBC), hemoglobin (HB), hematocrit (HCT), and total protein (TP) decrease in groups exposed to Cd and Pb. The cortisol and glucose levels exhibit a more noticeable (P < 0.05) increase with prolonged exposure to Cd and Pb than the control group. On day 30, the survival rate decreased more in the Pb-exposed group. The findings of this study indicate that exposure to sublethal doses of Cd and Pb induces stress in Wallagu attu, resulting in rapid changes in specific hemato-biochemical parameters.

Assessment and Bioaccumulation of Heavy Metals in Water, Fish (wild and Farmed) and Associated Human Health Risk.

Worldwide, anthropogenic activities are causing the natural environment and aquaculture systems to become heavily contaminated with heavy metals, which can lead to consumer's health problems. In the current study, wild and farmed fish (n = 30) and water samples (n = 6) have been collected from the Chashma barrage and fish farm to assess the heavy metals concentration, i.e., Cu, Cd, Pb, Zn and Cr, in the water and some important organs (gills, liver, muscle, brain and bones) of wild and farmed fish (Labeo rohita) using Graphite furnace Atomic absorption spectrometry. Bioaccumulation factor and human health risk assessment were calculated to measure the health status of both fish and humans. Results show that in wild and farm fish's gills, muscles and bones, the trend of the heavy metals was Zn > Pb > Cu > Cd > Cr. On the other hand, the brain and liver show Zn > Cu > Pb > Cd > Cr trend. Comparatively, the heavy metals concentration was mostly higher (P < 0.05) in wild fish. Further; in both fish habitats (water) the heavy metals (Cd and Pb) concentration was higher than the WHO standard level, while in the body, Cd was higher (P < 0.05) in all studied organs except the muscle, Cr was only lower (P > 0.05) in muscle and brain. Pb was higher (P < 0.05) in all studied organs of both fish. Bioaccumulation of heavy metals was mostly higher (P < 0.05) in wild fish than in farmed. EDI and THQ were higher in wild fish, but the HI value was lower than 1 for both fish. Moreover, the PCA analysis suggests a positive correlation between heavy metals concentration in fish organs and the water of both fish (wild and farmed). Results indicated that farmed fish showed less potential risk to humans than wild fish.

Seasonal Variation and Association of Heavy Metals in the Vital Organs of Edible Fishes from the River Jhelum in Punjab, Pakistan.

The toxic effects of heavy metals are drastic, including accumulation. Fish species are important bio-indicators of heavy metal pollution in aquatic bodies. The current study aimed to assess the seasonal variation of heavy metals in the vital organs of mostly consumed fishes in River Jhelum, Pakistan. Samples of fish, including Wallago attu (Malhi), Rita rita (Khagga), and Mystus seenghala (Singhari), were collected from four different sites, i.e., Khushab, Muhammad Wala (M. Wala), 8.R.D and Rasool barrage during summer and winter seasons. Heavy metals such as iron (Fe), lead (Pb), chromium (Cr), cobalt (Co) and Cadmium (Cd) were estimated through acid digestion and spectrometric analysis. Results showed a significantly higher (P < 0.05) amount of these metals in the liver, followed by the kidneys of fish species. There were seasonal variations in the absorption of these metals as well. Cr (11.71) and Fe (58.66) were detected in higher amounts in Khagga which showed the greatest affinity for certain metals in some cases. In contrast, Singhari showed the greatest affinity to other metals in other cases. Comparative analysis revealed that there was a highly significant (P < 0.05) difference for the accumulation of almost all metals in both seasons and summer had the highest concentration of Cd, Pb, Co, Cr and Fe as compared to winter in all four sampling stations in the case of kidney and liver of all the three fishes. Elevated levels of heavy metals were detected in the summer due to increased temperature. Heavy metals found in the River Jhelum may demonstrate that metals can significantly affect the fish species.

Human Health Risk Assessment due to the Incidence of Heavy Metals in Different Commercial Feeds Used for the Culturing of Biofloc Fish (Nile tilapia: Oreochromis niloticus).

Fish meat is a major and rich source of white protein; its quality is determined by the fish feed. However, the low-quality feed may contribute to a source of contamination if it does not fulfill the standard protocol. Biofloc is considered one of the most efficient, successful aquacultures, but this system is still under investigation for its efficiency and safety. Thus, current study focused on the heavy metal contamination in biofloc fish fed on different commercial feeds and human health risk analysis. Samples of extensively used three feeds (Supreme™, Hitech™, and MH-Aqua™), tanks water, and biofloc fish (gills, liver muscle) were collected for heavy metals (Cu, Cd, Pb, and Cr) analysis using atomic absorption spectrometry. An experiment was designed by dividing the fish into three groups: group 1 (Supreme™), group 2 (Hitech™), and group 3 (MH-Aqua™). A bioaccumulation factor and human health risk assessment have been calculated to measure fish and human health. Results revealed that most of the heavy metal concentration was higher (P < 0.05) in MH-Aqua™ feed compared to others. Similarly, heavy metal concentration was higher (P < 0.05) in the water of group 3, where fish was cultured on MH-Aqua™ feed. However, in the fish gills, liver, and muscle, the heavy metal concentration was significantly greater in group 3 fed on MH-Aqua™ feed, followed by group 1. Heavy metals in all feeds were positively correlated to the heavy metal concentration of the fish muscles. The bioaccumulation factor for Cu and Pb was higher in the fish liver, Cd and Cr in the case of fish gills, and least in the fish muscle. EDI and THQ values vary in all the groups, while the HI value was found lower than 1 in group 1 and group 2 but higher in group 3 fed on MH-Aqua™ feed. Strict checks and balances in formulating a diet will be helpful to progressively lower the amount of dangerous heavy metals.

Evaluation of the antibacterial activity and protein profiling of Nile tilapia (Oreochromis niloticus) epidermal mucus under different feeds and culture systems (biofloc technology and earthen pond).

The mucus layers of fish serve as the main interface between the organism and the environment. They play an important biological and ecological role. The current study focuses on Nile tilapia epidermal mucus reared under different commercial feeds (coded A and B) and environments (biofloc technology and earthen pond systems). Crude protein levels in feed A and B were 30% and 28%, respectively. Water parameters in all culturing systems were suitable for tilapia throughout the study period. The antimicrobial potency of tilapia (n = 5 from each) epidermal mucus was tested in vitro against human and fish pathogenic strains viz. Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Francisella noatunensis, and Aeromonas hydrophila. To determine the antimicrobial activity, zones of inhibition (ZOI) were measured in millimetres and compared with two antibiotics (chloramphenicol and ciprofloxacin). SDS-PAGE analysis was performed on skin mucus samples of tilapia to determine protein quantity and size (molecular weight). Results of tilapia skin mucus (crude and aqueous) revealed a strong antibacterial effect against all the selected pathogenic strains. However, variation has been observed in the mucus potency and ZOI values between the biofloc and pond tilapia mucus. The crude mucus of tilapia fed on feed A and cultured in the pond exhibited strong antibacterial effects and high ZOI values compared to the mucus of biofloc tilapia, aqueous mucus extracts and positive control chloramphenicol (antibiotic). The SDS-PAGE results showed that the high molecular weight proteins were found in the collected epidermal mucus of BFT-B (240 kDa) and EP-B (230 kDa). Several peptides in fish skin mucus may play a crucial role in the protection of fish against disease-causing pathogens. Thus, it can be utilized in the human and veterinary sectors as an 'antimicrobial' for treating various bacterial infections.

Cobalt Uptake by Food Plants and Accumulation in Municipal Solid Waste Materials Compost-amended Soil: Public Health Implications.

One of the most pressing environmental issues is how to properly dispose of municipal solid waste (MSW), which represents both a substantial source of concern and a challenge. The current study evaluated cobalt (Co) accumulation in MSW, their uptake by different vegetables grown for two years, and related human health risks. Vegetables were grown in four different groups, such as one control (ground soil), and the remaining treatment groups (T1, T2, and T3) received varying concentrations of MSW. The analysis of Co was done through an atomic absorption spectrophotometer (AAS). Results revealed that the concentration of Co was higher in all the vegetables (n = 15) grown in soil supplemented with 75% MSW during 2nd growing year. Among all vegetables, the highest concentration of Co was observed in Solanum tuberosum at T3 during 2nd growing year. The pollution load index (PLI) value for vegetables during both growing years was more than 1 except in control soil. The findings indicated that the highest enrichment factor (EF) and hazard resilience index (HRI) value of 0.09 was present in S. tuberosum. Health index values for cobalt in the study were below 1. The HRI < 1 indicated that consumers do not face any immediate health risks. The investigation of Co concentrations in blood samples obtained from individuals residing in different areas contributes a human health perspective to the research. The findings indicate that the concentration of Co rises with an increasing proportion of MSW. While the metal levels in MSW-treated soil were not high enough to classify the soil as polluted, the results recommend that recycling MSW can substitute mineral fertilizers. Nevertheless, the presence of cobalt in MSW may directly affect soil fertility and could impact crop production and human health.

Investigation of the incidence of heavy metals contamination in commonly used fertilizers applied to vegetables, fish ponds, and human health risk assessments.

Overuse of fertilizers on agricultural lands and fish ponds may result in serious pollution problems, such as heavy metals that can enter the food chain and pose serious health problems. Due to this, the present study investigates the incidence of heavy metals in commonly used fertilizers and its association with heavy metals in vegetables, soil, fish species, and pond water. Samples were collected from different sites (fields and ponds) in district Kohat, where the application of fertilizers was common and control groups (no fertilizers used). Heavy metal analysis was carried out through a spectrophotometer. Results showed higher Cd and Cr concentrations in triple superphosphate (TSP), Cu and Pb in nitrogen, phosphorus, and potassium (NPK), while lower concentrations were found in gypsum. In vegetables (onion, tomato, brinjal, and potato) and associated soil, most of the heavy metals concentrations were significantly higher (P < 0.05) in fertilizer-applied sites than in the control. Also, the Cd concentration in potatoes and Pb level in all vegetables obtained from sites were greater than the WHO/FAO standard limit. In the case of fish species (Hypophthalmichthys molitrix and Cyprinus carpio) muscles and their habitat (water), all the understudy heavy metals were notably higher (P < 0.05) in fertilizer-applied sites (ponds) than the control group. Collectively, in all vegetables and muscles of fish species, the bioaccumulation factor was higher in sites compared to the control. The estimated daily intake (EDI) and target hazard quotient (THQ) values were also higher in fertilizer-applied sites (fields and ponds) than control. The health index (HI) value was > 1 in vegetables (onion, tomato, and potato) and fish muscles collected from different sites compared to the control. Thus, there is the possibility of severe health risks. The use of fertilizers must be carefully monitored in order to ensure that humans and animals are safe from exposure to heavy metals.

Withania coagulans fruit extract: a possible useful additive in ameliorating growth and immunity of Labeo rohita (Hamilton, 1822).

This study aimed to assess the Withania coagulans fruit extract's effects on growth and haematological parameters of Labeo rohita. Healthy fish (n = 120) were divided in four groups. Experimental diet was prepared with different extraction of W. coagulans (0%, 1%, 1.5% and 2%). Diet was provided for eight weeks to all groups except the control group which fed on basal diet. The 1.5% and 2% W. coagulans extracts lead improvement in feed conversion ratio and specific fish growth rate; haematological parameters in fish group fed with 2% of W. coagulans showed increased in WBC, RBC, Hb, MCHC, PCV. The values of plasma proteins and lysozyme activity were higher in the experimental group compared control group. Fish fed with W. coagulans extracts showed increased of immunity and less mortality respect to others. It suggested that W. coagulans could be considered valuable support for the improve growth and immunity in Labeo rohita.

Assessment of Genetic Damage in Coal Miners of Punjab, Pakistan.

Coal miners are continuously exposed to coal mine dust and airborne particulate that act as a potential risk to their health. The present study evaluates the DNA damage in coal miners using the Buccal Micronucleus Cytome (BMCyt) assay. The samples of the blood and buccal epithelial cells of 40 coal miners and 20 control subjects were taken from coal mines of Pail and Padhrar, Pakistan, to establish buccal anomaly frequencies of metal levels in the blood. Besides this, work history and duration hours were also analyzed. Results revealed that micronucleus frequencies positively correlated with the metal concentrations in the miner's blood. The change in the extent of nuclear damage per unit change in the year was 0.170 for micronuclei; however, with addition in each year of working experience, nuclear buds and broken egged nuclei increased by 0.316 and 0.194 units, where each year increases karyolysis by 0.349 units and karyorrhexis by 0.308 units, respectively. An increase in work hours and working years was positively correlated with cytogenetic damage. Nuclear damage in coal miners due to occupational exposure is obvious and increases with increasing work experience. Hence, the Buccal Micronucleus Cytome assay has proved to be an effective cytogenetic biomonitoring tool for assessing genetic and nuclear damage in coal miners.

Evaluation and Association of Heavy Metals in Commonly Used Fish Feed with Metals Concentration in Some Tissues of O. niloticus Cultured in Biofloc Technology and Earthen Pond System.

For successful aquaculture, the primary need is the quality of fish feed, which determines fish flesh quality. The current study was conducted to evaluate the number and concentration of heavy metals in commonly used fish feeds and fish gills, liver, and muscle of biofloc technology and earthen pond systems. Besides this, the correlation between heavy metals in fish feeds with detected metals in the gills, liver, and muscle of fish was also determined. Results revealed that heavy metals concentration, including Cu and Cd, in feed B was significantly greater than in feed A, but the Zn level in feed A was significantly higher than in feed B. Furthermore, the concentration of heavy metals in fish of both aquaculture systems was significantly higher in the liver than in the gills and muscle. The metal concentration in fish feeds and fish edible parts (muscle) was lower than the WHO standard level; however, the amount of Pb was higher in the fish muscle, which is harmful for human consumption. Though the correlation test revealed that all of the metals from the feeds were positively correlated to the metals detected in the fish, but most of the estimated correlation was insignificant. From the current study, it can be concluded that the fish feed producers need to measure feed quality adequately to avoid hazardous contamination by heavy metals in the feed. The ultimate consumer, fish and humans, may, otherwise, be predisposed to assimilate and accumulate these heavy metals.

Assessment and Bioaccumulation of Heavy Metals in Fish Feeds, Water, and Some Tissues of Cyprinus carpio Cultured in Different Environments (Biofloc Technology and Earthen Pond System).

Fish feed quality is the main determinant of fish flesh quality, so it is important for successful aquaculture. The current study determines the concentration of heavy metals in fish feeds (A and B), water, and their bioaccumulation in gills, liver, and muscle of C. carpio cultured in different environments (biofloc technology and earthen pond systems). In addition, the correlation between heavy metals in fish feeds with bioaccumulated metals in fish tissues was also determined. Results revealed that most heavy metal concentration was significantly greater (P < 0.05) in feed B than in feed A but in permissible range, while all the heavy metal concentration was notably higher in earthen ponds than in biofloc technology. Result from the bioaccumulation factor and concentration of the metals showed that heavy metals were highly accumulated in the fish liver followed by gills. The metal concentration in fish feeds and fish edible parts (muscle) was lower than the WHO standard level; however, the amount of Pb was higher in the fish muscle, liver, and gills, which is harmful for human consumption and also for fish health. Though the correlation test revealed that all of the metals from the feeds were positively correlated to the metals in fish tissues, but most of the estimated correlation was significant and linearly correlated. It can be concluded that producers must measure feed quality correctly to avoid heavy metal contamination because it may assimilate and accumulate in the food chain.

Cadmium Sub-Lethal Concentration Effect on Growth, Haematological and Biochemical Parameters of Mystus seenghala (Sykes, 1839).

The current study aims to assess the cadmium sub-lethal concentration influence on growth and haematological and biochemical parameters of Mystus seenghala. A total of 60 fish of three different length groups (20 each) were collected from Head Qadirabad, Pakistan. The fish were treated to the sub-lethal concentration viz. one-third of LC50, for 16 weeks except for the control groups. Water quality parameters were kept constant during the entire course of the research, and the major parameters were measured as temperature (28.03 ± 0.03 °C), DO (5.82 ± 0.14 mg L-1), pH (8.00 ± 0.01) and total hardness (249.98 ± 0.01 mg L-1). Findings revealed that the growth of three treated variant length groups was affected negatively by cadmium exposure and showed significantly (P < 0.05) lower average wet weight, body length and condition factor as compared to control groups, while the feed conversion ratio (FCR) increases by increasing the exposure duration. The haematological parameters including values of Hct, Hb and MCHC were significantly (P < 0.05) reduced in all Cd-treated groups than control groups, whereas the level of MCH and MCV were significantly higher, but no significant difference was found in the value of RBCs in all treated groups. Biochemical parameters such as ALT, AST, total lipid and glucose level in Cd exposure groups were significantly higher, while the total protein level was significantly (P < 0.05) reduced in all treated groups as compared to control groups. From the current study, it has been concluded that the growth, haematology and biochemical parameters are important indicators of ecotoxicology particularly contamination of the cadmium and health of the fish.

Effect of fortified feed with olive leaves extract on the haematological and biochemical parameters of Oreochromis niloticus (Nile tilapia).

Olive leaves have been used traditionally to improve the immunity against several infections. Current study aimed to examine the effect of fortified feed with olive leaves extract on haematological and biochemical parameters of Oreochromis niloticus. Total 400 healthy fish were divided randomly into four groups and were cultured for two months in glass tanks of 250 L water capacity in triplicate. Fortified feed was prepared with various extract, 1%, 1.5% and 2% concentration in diet, while, control-group diet was without extraction. Results showed that all the haematological parameters including WBC, RBC, Hb, PCV and plasma protein were significantly increased while the biochemical parameters such as globulin albumen ratio increased in fish which fed on 1% of extract, but the level of cholesterol and triglycerides was decreasing by increasing the concentration of extract. It was concluded that olive leaves extract can be used to enhance the immunity, growth and health of fish.

Effect of Fortified Feed with Phyto-Extract on the First Physical Barrier (Mucus) of Labeo rohita.

The aim of the current study was to assess the effect of two different fortified feeds with different concentrations of two important medicinal plants (Withania coagulans and Zingiber officinale) on the mucosal immunity of Labeo rohita. After a dietary intervention, mucus was tested against five pathogenic bacteria (in-vitro), while experimental fish were tested against the ectoparasite (Lernaea) (in-vivo). Our results revealed that all fish groups fed with different concentrations (1, 1.5, and 2%) of Z. officinale had low molecular weight proteins and did not develop any significant signs of parasitic infection, with low mortality rate; whereas the groups that were fed with W. coagulans (particularly with 1% and 2%), including a control group, developed rapid signs of infection with high mortality rate. The highest hemagglutination titer value was recorded for the fish fed with 1% and 1.5% of Z. officinale. The lowest value was found for the fish fed with 2% of W. coagulans. The mucus of all fish of fortified groups was active and inhibited the growth of tested bacterial pathogens as compared to the control group. Further, Z. officinale groups showed greater efficacy against bacteria as compared to the W. coagulans groups. In conclusion, Z. officinale can be considered as a potential and functional ingredient in aquaculture feed. Furthermore, future studies should be conducted to investigate more details on the subject.