Acute toxicity of palm oil mill effluent on zebrafish (Danio rerio Hamilton-Buchanan, 1822): Growth performance, behavioral responses and histopathological lesions.

Evaluating the toxicity of Palm Oil Mill Effluent (POME) is critical as part of the effort to develop waste management regulations for the palm oil industry. In this study, we investigated the acute toxicity of POME on growth performance, behavioral response, and histopathology of gill and liver tissues of zebrafish (Danio rerio). In total, 550 adult male zebrafish were used for the toxicity experiment including range finding test, acute toxicity test, growth performance and behaviour test. Static non-renewal acute toxicity bioassays were conducted by exposing fish to POME (1.584-9.968 mL/L) for 96 h. Growth performance, behavior response, and histopathological lesions in untreated and POME treated (96-h LC50: 5.156 mL/L) fish were measured at 24, 48, 72 and 96 h. Time-dependent significant decline in body length and body weight of POME-exposed zebrafish was observed. Furthermore, several behavioral changes were recorded, including hyperactivity, loss of balance, excessive mucus secretion, and depigmentation. Decreasing operculum movement and oxygen consumption rate as well as alterations in gill tissues (i.e. hyperplasia, hypertrophy, hemorrhage, and necrosis) of POME-exposed zebrafish were observed, suggesting a dysfunction in respiratory performance. On the other hand, liver tissue alterations (congestion, hemorrhage, hyperplasia, shrinkage of hepatocytes, hydrophilic degeneration, and necrosis) indicated a disruption in detoxification performance. We conclude that exposure to POME at acute concentration caused histopathological lesions both in gill and liver tissue along with changes in fish behaviors which disrupted respiratory and detoxification performance, resulting in mortality and reduced growth of zebrafish. These findings might provide valuable information for guiding POME management and regulation.

Investigations of hemato-biochemical and histopathological parameters, and growth performance of walking catfish (Clarias batrachus) exposed to PET and LDPE microplastics.

Fish inhabiting various trophic levels are affected differently as the presence of microplastic (MP) in the water column and their ingestion by fish varies. Walking catfish (Clarias batrachus) inhabits the bottom of the water bodies. To understand the effects of MP, we exposed C. batrachus to two types of MP - polyethylene terephthalate (PET) and low-density polyethylene (LDPE) for 60 days. After exposure, hematological indices, mainly red blood cells and hemoglobin levels decreased, and white blood cells increased significantly compared to the control group (p < 0.05). A significant increase in the levels of blood urea and glucose was observed, and serum glutamic pyruvate transaminase and serum glutamyl oxaloacetic transaminase activity remained elevated (p < 0.05). Histopathological examination of the liver, kidney, intestine, and gills showed morphological alterations. Moreover, MP exposure caused growth retardation (p < 0.05) in C. batrachus. Widespread pollution of water bodies by MP may impose serious ecological risks to bottom-feeding fish in Bangladesh.

Dried fish more prone to microplastics contamination over fresh fish - Higher potential of trophic transfer to human body.

Globally, microplastics (MPs) contamination in aquatic organisms is emerging as an alarming phenomenon. In the present study, we investigated MPs in three commercially important fishes (Bombay duck Harpadon nehereus, ribbon fish Trichiurus lepturus and hairfin anchovy Setipinna phasa) in fresh and dried conditions collected from two sites (Chattogram and Kuakata) of the Bay of Bengal. It was evident that fresh T. lepturus ingested highest amount of MPs through the gills (6.41 mps/g) from Chattogram followed by in the gastrointestinal tract, GIT (6.20 mps/g) and in the muscle (1.20 mps/g) from Kuakata. Among the fresh fishes, H. nehereus from Kuakata accumulated highest amount of MPs (0.21 mps/g), while S. phasa from Kuakata contained the least amount of MPs (0.06 mps/g). On the other hand, among the dried fishes, T. lepturus from Kuakata contained highest amount of MPs (46.00 mps/g), while S. phasa from Kuakata retained lowest amount of MPs (2.17 mps/g). Strangely, all the dried fishes showed significantly higher amount of MPs compared to fresh fishes from both the locations. Fiber was the most dominant type of shape of MPs which accounted 66 %, followed by fragment (27.38 %), microbeads (3.59 %), film (1.48 %), foam (1.31 %) and pellet (0.25 %). Size-wise, the major portion (39.66 %) of MPs was present to be in size range less than 0.5 mm followed by 37.67 % in the size range of 0.5-1.0 mm group and rest 22.67 % within 1.0-5.0 mm. Red (41.55 %) colored MPs was the most prominent, followed by brown (22.11 %), blue (16.32 %), pink (11.69 %), purple (5.10 %), and green (2.25 %). Among polymer types, low-density polyethylene (LDPE) was the most common (38 %), followed by polystyrene (PS-22 %), polyvinyl chloride (PVC-16 %), polyamide (PA-13 %) and ethylene-vinyl acetate (EVA-9 %). The present study confirms high occurrence of MPs in the dried fishes over the fresh fishes from the Bay of Bengal, with high potential of trophic transfer to the human body.

Microplastics pollution in the river Karnaphuli: a preliminary study on a tidal confluence river in the southeast coast of Bangladesh.

Bangladesh is a deltaic country in Asia, and its riverine systems ultimately drain into the Bay of Bengal. Plastic is a severe environmental issue for coastal-marine ecosystems due to the indiscriminate usage and discarding of plastic items in the upstream river that eventually find their route into the Bay of Bengal. Microplastics (MPs) are widespread pollutants in almost all environmental compartments, including aquatic environments. This study aimed to quantify and understand the distribution of microplastics in surface water and sediments of the river Karnaphuli, a tidal confluence river adjacent to the Chattogram seaport city of Bangladesh, a highly inhabited and industrial area on the southeast coast of the Bay of Bengal. A manta trawl net (300-µm mesh size) was used to collect surface water samples, while an Ekman dredge was used to collect sediment samples. The concentrations of microplastics in the surface water of the river Karnaphuli during late monsoon, winter, and early summer were recorded to be 120,111.11, 152,222.22, and 164,444.44 items/km2, respectively, while in sediments, those were recorded to be 103.83, 137.50, and 103.67 items/kg, respectively. A higher abundance of microplastics was observed in downstream surface water (228,888.88 items/km2) and sediments (164.17 items/kg). Smaller sizes (0.3 to 0.5 mm) of microplastics were predominant, fibers or threads were the frequent types, and black was the most common color in the river Karnaphuli. The Fourier transform infrared analysis revealed that polyethylene terephthalate (surface water: 22%, sediments: 19%), polyamide (surface water: 15%, sediments: 13%), polyethylene (surface water: 12%, sediments: 18%), polystyrene (surface water: 13%, sediments: 11%), and alkyd resin (surface water: 13%, sediments: 10%) were the most prevalent polymers in the river Karnaphuli. Moreover, there was a moderate positive correlation between MPs abundance in surface water and sediments. Therefore, improved long-term research (in different seasons with horizontal and vertical monitoring) is necessary in order to accurately determine the flux of microplastics from the river Karnaphuli to the Bay of Bengal.

Application of fenitrothion on Heteropneustes fossilis causes alteration in morphology of erythrocytes via modifying hematological parameters.

In Bangladesh, the extensive use of fenitrothion on crops and in aquaculture ponds inevitably threatens a range of aquaculture species, including fish, owing to stress responses and physiological disturbances. The present study elucidated the potential toxic effects of fenitrothion on the blood biomarkers (haemato-biochemistry and structure of erythrocytes) of stinging catfish (Heteropneustes fossilis), a commercially significant aquaculture species. Fish were exposed to four sub-lethal concentrations (0%, 10%, 20%, and 40% of the 96-h LC50 value) of fenitrothion in triplicate and observed on the 7th, 14th, 21st, and 28th day following exposure. With increasing fenitrothion concentration, blood glucose and white blood cell levels increased significantly; in contrast, hemoglobin, red blood cell, and packed cell volume substantially decreased. However, the mean corpuscular volume and mean corpuscular hemoglobin did not change significantly during the exordial period (0-7 d); although, at a later stage, changes were observed. Frequencies of observed erythrocytic nuclear abnormalities, such as degeneration, bi-nucleus, micronucleus, notch nucleus, and nuclear bridge and erythrocytic cellular abnormalities, such as echinocytes, fusion, elongation, and tear drop morphology increased significantly in a concentration-dependent manner. Differences between the control individuals and those individuals under treatment were considered insignificant for twin cells on the 14th day of exposure. The study showed the pernicious impact of the effects of fenitrothion on H. fossilis through physiological alteration, which is likely to pose challenges for aquaculture production.

Thiamethoxam affects the developmental stages of banded gourami (Trichogaster fasciata).

Thiamethoxam (THM), a type III systemic neonicotinoid insecticide, is widely used in agriculture in many countries, including Bangladesh. THM can enter the aquatic systems through the runoff, spray-drift and groundwater leaching and can affect the aquatic organisms, including fish. Current environmental levels of THM in Bangladesh waters are variable. However, the presence of this chemical in the aquatic environment and its possible effects on the fish inhabiting those water bodies is concerning. To understand the effects of environmental THM on the development of embryo and larvae, the present study used banded gourami (Trichogaster fasciata), a freshwater fish species distributed in different Asian countries, including Bangladesh. In laboratory setting, fertilized eggs (n = 100) and one-day-old banded gourami larvae (n = 100) were exposed to six concentrations of THM (0, 0.02, 0.2, 2, 20, 200 mg/L) in three replicates, in which three concentrations were within the environmentally relevant levels. Hatching rate, incubation period, mortality, and malformations of embryo and larvae were observed. The hatching success and survival of embryo and larvae significantly decreased with increasing THM concentrations. The 24-h LC50 of THM for the embryo was 4.24 mg/L. The 24-h, 48-h, 72-h, and 96-h LC50 values of THM for one-day-old larvae were 12.20, 3.80, 0.78, and 0.27 mg/L, respectively. Overall developmental malformations included lordosis, notochord abnormality, yolk-sac edema, dark brown yolk sac, hemorrhage, and irregular caudal fin. These abnormalities in embryos were common across all the concentrations of THM applied. The results of the present study suggest that environmentally relevant concentrations of THM can induce developmental defects in the embryo and larvae of banded gourami.

Alterations in hematological parameters and the structure of peripheral erythrocytes in Nile tilapia (Oreochromis niloticus) exposed to profenofos.

The burden of pesticide use from agricultural intensification lies in the fact that pesticides may end up in aquatic ecosystems and have pernicious effects on non-target organisms, including fish. Different blood biomarkers, including hemato-biochemical indices, erythrocytic nuclear abnormalities (ENA), and erythrocytic cellular abnormalities (ECA), were observed in Nile tilapia (Oreochromis niloticus) after exposure to varying sub-lethal concentrations (0%, 5%, 10%, 20%, and 40% of 96-h LC50) of profenofos at different time intervals (7, 14, 21, and 28 days). The results revealed that glucose and white blood cell (WBC) levels significantly increased, while hemoglobin, red blood cell (RBC), and packed cell volume (PCV) significantly decreased in a time- and concentration-dependent manner. Aberrant erythrocytic morphology-derived ENA, such as nuclear degeneration, micronuclear formation, binuclear development, nuclear budding, and karyopyknosis, significantly increased with time in profenofos-exposed groups compared to controls. Between the treatment and control groups, a significant execution was discerned for teardrop and fusion type ECA. For other cellular aberrations of erythrocytes, including elongated, twin, and spindle, a significant difference appeared only at the beginning of the experiment (day 7). This study concludes that the presence of widely used profenofos in aquatic systems has a pernicious effect on Nile tilapia.

Exposure to Lead Nitrate Alters Growth and Haematological Parameters of Milkfish (Chanos chanos).

To date, the effects of lead on the growth and haematological parameters of milkfish are still not well understood. For this reason the present study seeks to explain this conundrum. Sub-adult female milkfish were exposed to four concentrations of lead nitrate (0 mg/l, 42.64 mg/l, 63.97 mg/l and 85.29 mg/l) for 40 days. Results revealed that exposure to lead nitrate caused significant changes in growth and haematological parameters of milkfish. Weight gain, length gain, specific growth rate, feed efficiency, and feed conversion ratio of milkfish declined significantly at the highest concentration treatment. Moreover, lead nitrate exposure significantly decreased the value of red blood cells, hemoglobin, hematocrit, and mean corpuscular hemoglobin concentration, along with a marked increase in mean corpuscular volume and mean corpuscular hemoglobin. Several erythrocyte malformations to cells including swelling, deformation, doubling, binucleus, laceration of the membrane, hemolyzation and vacuolation were all observed.

Toxic effects of fenitrothion on freshwater microcosms in Bangladesh.

The organophosphate pesticide fenitrothion is widely used as an agricultural pesticide to control tiger bug in larval rearing for aquaculture. The objectives of the present study were to assess the effects of fenitrothion on certain structural (phytoplankton, zooplankton, macro-invertebrates and periphyton) and functional (organic matter decomposition) endpoints of freshwater microcosms. Fenitrothion 50 EC was applied in 12 microcosms (PVC tanks having 400 L of dechlorinated tap water) providing concentrations of 0, 25, 50 and 100 µg/L at a 4-day interval over a period of 4 weeks. Each of the experimental treatment was executed in three replicates. The results indicated the consistent significant effects for most of the species composition of zooplankton and macro-invertebrates. Univariate analysis showed a significant decrease in abundance (p < 0.05) of all identified insects (i.e. Notonecta sp., Gerris sp., Ranatra linearis and Chironomid larvae), when compared to control in all sampling days throughout the treatment period (no observed effect concentration; NOEC = < 25 µg/L). No consistent significant effects were observed for most of the phytoplankton taxa and organic matter decomposition and water quality variables (dissolved oxygen, free CO2, pH, nitrate etc.). However, several taxa of different endpoints were found sensitive to even the lowest concentration of fenitrothion (25 µg/L). Further studies with acute and chronic conditions are recommended involving more local species exposed to < 25 µg/L of fenitrothion.

Toxicity of palm oil mill effluent on the early life stages of Nile tilapia (Oreochromis niloticus, Linnaeus 1758).

Harmful effects of several pollutants have been reported on early life stages of fish. However, the effects of palm oil mill effluent (POME) on fish early life stages are still unexplored. Therefore, the objective of this present study was to elucidate the impact of POME on the early life stages of Nile tilapia (Oreochromis niloticus). Fertilized eggs of Nile tilapia were exposed to four concentrations of POME (0, 1.565, 2.347, and 3.130 mg/L) in 20 plastic funnels. Each of the control and treatment groups was maintained in five replicates. The cumulative hatching rate, malformation rate, body length, and deformities of larvae were analyzed. Results showed that hatching rate and survival rate of Nile tilapia larvae significantly decreased with increasing concentrations of POME. In contrast to, malformation rate and heart rate were significantly increased. Furthermore, results showed several malformations of Nile tilapia larvae including lordosis, kyphosis, and curved tail when exposed to 1.565 mg/L, 2.347 mg/L, and 3.130 mg/L of POME concentrations. Further research is required to understand the physiological mechanisms of different endpoints in the early stages of Nile tilapia induced by the toxicity of POME.

Effects of long-term chlorpyrifos exposure on mortality and reproductive tissues of Banded Gourami (Trichogaster fasciata).

This study assessed the long-term toxicity of chlorpyrifos on survival and reproduction of Banded Gourami by using mortality, gonado-somatic index (GSI) and histopathological observations as endpoints. Adult fish were exposed to five different concentrations of chlorpyrifos (0, 15, 50, 150, 500 µg/L) in 15 PVC tanks for 15, 30, 45, 60 and 75 days. Results showed that all male and female fish died after 15 days of 500 µg/L chlorpyrifos exposure. No consistent significant effect was observed for both male and female GSI. Furthermore, results showed dose- and time-dependent histopathological alterations for both ovary and testes. The 60-d No Observed Effect Concentration (NOEC) for most histopathological alterations of Banded Gourami ovary and testes was 50 µg/L, while 60-d NOEC for mortality of both male and female fish was < 15 µg/L. The results show that the long-term exposure to chlorpyrifos not only affect the reproductive tissues of Banded Gourami at exposure concentrations but also cause their mortality. Future studies should evaluate effects at lower concentrations.

Environmental monitoring and risk assessment of organophosphate pesticides in aquatic ecosystems of north-west Bangladesh.

The use of organophosphate pesticides (OPPs) to protect a variety of crops has increased in Bangladesh. OPPs may contaminate surrounding aquatic environments through several routes including spray drift, surface runoff and groundwater leaching. Since it is unknown how much OPP end ups in aquatic environment in Bangladesh, the objectives of the present study were to quantify the residues of ten most commonly used OPPs in water and sediment of water bodies of north-west Bangladesh and to assess their ecological risks for aquatic organisms. The risks of the pesticides in surface water and sediment were assessed using a first-tier risk quotient (RQ) approach. The higher-tier PERPEST model was used to refine the ecological risks of pesticides when RQ indicated a potential risk. Results showed the most frequently detected pesticides that appeared in high concentrations were chlorpyrifos, diazinon and quinalphos in surface water and sediment. The highest concentration of OPPs measured in water was 9.1 µg chlorpyrifos/L (median of 1.95 µg/L), while this was 51 µg diazinon/kg dw (median of 11 µg/kg dw) for sediment. Furthermore, results showed high acute and/or chronic RQs (RQ > 1) in surface water and sediment for chlorpyrifos, diazinon, quinalphos, malathion and fenitrothion. The higher-tier PERPEST model confirmed risks of chlorpyrifos, diazinon, quinalphos and fenitrothion for aquatic insects, micro- and macro-crustaceans which were previously derived by RQ-based risk assessment for aquatic organisms. Furthermore, the results of the PERPEST model also indicated possible indirect effects of these pesticides on algae and macrophytes, community metabolism, rotifers and other macro-invertebrates.

Effects of imidacloprid on the ecology of sub-tropical freshwater microcosms.

The neonicotinoid insecticide imidacloprid is used in Bangladesh for a variety of crop protection purposes. Imidacloprid may contaminate aquatic ecosystems via spray drift, surface runoff and ground water leaching. The present study aimed at assessing the fate and effects of imidacloprid on structural (phytoplankton, zooplankton, macroinvertebrates and periphyton) and functional (organic matter decomposition) endpoints of freshwater, sub-tropical ecosystems in Bangladesh. Imidacloprid was applied weekly to 16 freshwater microcosms (PVC tanks containing 400 L de-chlorinated tap water) at nominal concentrations of 0, 30, 300, 3000 ng/L over a period of 4 weeks. Results indicated that imidacloprid concentrations from the microcosm water column declined rapidly. Univariate and multivariate analysis showed significant effects of imidacloprid on the zooplankton and macroinvertebrate community, some individual phytoplankton taxa, and water quality variables (i.e. DO, alkalinity, ammonia and nitrate), with Cloeon sp., Diaptomus sp. and Keratella sp. being the most affected species, i.e. showing lower abundance values in all treatments compared to the control. The observed high sensitivity of Cloeon sp. and Diaptomus sp. was confirmed by the results of single species tests. No significant effects were observed on the species composition of the phytoplankton, periphyton biomass and organic matter decomposition for any of the sampling days. Our study indicates that (sub-)tropical aquatic ecosystems can be much more sensitive to imidacloprid compared to temperate ones.

Toxicity of cypermethrin on the embryo and larvae of Gangetic mystus, Mystus cavasius.

The objective of the present study was to elucidate the effects of cypermethrin on the embryo and the larvae of Gangetic mystus, Mystus cavasius. Therefore, fertilized eggs (n = 100) and 1-day-old larvae (n = 100) were exposed to six different concentrations of cypermethrin (0, 2, 4, 8, 16 and 32 µg L-1) in each of the 18 plastic bowls. Each of the treatment and control was maintained in three replicates. The LC10 and LC50 values for Gangetic mystus embryos and larvae were calculated using probit analysis. Results showed the mortality of embryos significantly increased with increasing cypermethrin concentrations. The 24-h LC10 and LC50 (with 95% confidence interval) values of cypermethrin for embryo were 0.42 (0.14-0.81) and 5.60 (4.16-7.19) µg L-1, respectively. Hatching success decreased and mortality of larvae increased significantly with increasing cypermethrin concentrations. The 24-h LC10 and LC50 values (with 95% confidence limits) of cypermethrin for larvae were 1.72 (1.24-2.20) and 11.57 (10.09-13.42) µg L-1, respectively; the 48-h LC10 and LC50 for larvae were 1.34 (0.83-1.89) and 8.25 (6.87-9.91) µg L-1, respectively; the 72-h LC10 and LC50 for larvae were 1.13 (0.63-1.66) and 6.12 (4.91-7.47) µg L-1, respectively. Furthermore, results showed several malformations in embryos and larvae when exposed to the two highest concentrations of cypermethrin. The findings of the study suggest that 2 µg L-1 cypermethrin concentration in the aquatic environment may have deleterious effects on the development and the reproduction of Gangetic mystus.

Acute toxicity of chlorpyrifos to embryo and larvae of banded gourami Trichogaster fasciata.

This study elucidated the acute toxicity of chlorpyrifos on the early life stages of banded gourami (Trichogaster fasciata). To determine the acute effects of chlorpyrifos on their survival and development, we exposedthe embryos and two-day-old larvae to six concentrations (0, 0.01, 0.10, 1.0, 10 and 100 µg L-1) of chlorpyrifos in plastic bowls. Log-logistic regression was used to calculate LC10 and LC50 values. Results showed that embryo mortality significantly increased with increasing chlorpyrifos concentrations. The 24-h LC10 and LC50 values (with 95% confidence limits) of chlorpyrifos for embryos were 0.89 (0.50-1.58) and 11.8 (9.12-15.4) µg L-1, respectively. Hatching success decreased and mortality of larvae significantly increased with increasing concentrations of chlorpyrifos. The 24-h LC10 and LC50 values (with 95% confidence limits) of chlorpyrifos for larvae were 0.53 (0.27-1.06) and 21.7 (15.9-29.4) µg L-1, respectively; the 48-h LC10 and LC50 for larvae were 0.04 (0.02-0.09) and 5.47 (3.77-7.94) µg L-1, respectively. The results of this study suggest that 1 µg L-1 of chlorpyrifos in the aquatic environment may adversely affect the development and the reproduction of banded gourami. Our study also suggests that banded gourami fish can serve as an ideal model species for evaluating developmental toxicity of environmental contaminants.

Risk assessment of pesticides used in rice-prawn concurrent systems in Bangladesh.

The objectives of the current study were to determine the occupational health hazards posed by the application of pesticides in rice-prawn concurrent systems of south-west Bangladesh and to assess their potential risks for the aquatic ecosystems that support the culture of freshwater prawns (Macrobrachium rosenbergii). Information on pesticide use in rice-prawn farming was collected through structured interviews with 38 farm owners held between January and May of 2012. The risks of the pesticide use to human health were assessed through structured interviews. The TOXSWA model was used to calculate pesticide exposure (peak and time-weighted average concentrations) in surface waters of rice-prawn systems for different spray drift scenarios and a simple first tier risk assessment based on threshold concentrations derived from single species toxicity tests were used to assess the ecological risk in the form of risk quotients. The PERPEST model was used to refine the ecological risks when the first tier assessment indicated a possible risk. Eleven synthetic insecticides and one fungicide (sulphur) were recorded as part of this investigation. The most commonly reported pesticide was sulphur (used by 29% of the interviewed farmers), followed by thiamethoxam, chlorantraniliprole, and phenthoate (21%). A large portion of the interviewed farmers described negative health symptoms after pesticide applications, including vomiting (51%), headache (18%) and eye irritation (12%). The results of the first tier risk assessment indicated that chlorpyrifos, cypermethrin, alpha-cypermethrin, and malathion may pose a high to moderate acute and chronic risks for invertebrates and fish in all evaluated spray drift scenarios. The higher tier assessment using the PERPEST model confirmed the high risk of cypermethrin, alpha-cypermethrin, and chlorpyrifos for insects and macro- and micro-crustaceans thus indicating that these pesticides may have severe adverse consequences for the prawn production yields.