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Malathion toxicity: Effect on the liver of the fish Brachydanio rerio (cyprinidae)
Abstract
Four-month-old siblings of zebra fish, Brachydanio rerio, were exposed to 0.5, 0.7, 0.9, and 1.1 mg/liter of malathion for 7 days. DNA, RNA, protein, and free amino acid contents and the activities of acid and alkaline phosphatases were monitored in the liver. There was marked inhibition in DNA, RNA, and protein contents and the activities of acid and alkaline phosphatase. Total free amino acid content, however, significantly increased. Seven days withdrawal to freshwater indicated a recovery trend. The probable reasons for various changes are discussed.
... Singh and Sahai [47] also reported similar effects on kidney, liver and gills of malathion exposed Puntius ticto. Kumar and Ansari [48] observed degeneration of hepatic cells in Zebra fish exposed to 0.9 mg/L malathion for a period of four months and noted skeletal deformities. Barbhuiya and Dey [49] investigated the liver tissue of Heteropneustes fossilis exposed to 1.07 ppm (1/10th of 96h LC50) concentration of malathion for a period of 21 days and noted histological alterations with degeneration of hepatocytes and fibrous tissue and formation of vacuoles and appearance of macrophages. ...
... The fish had a progressive decrease in RNA and increase in DNA as exposure time increased. Kumar and Ansari [48] studied the DNA, RNA, protein, free amino acid contents and the activities of acid and alkaline phosphatases in zebra fish (Brachydanio rerio) exposed to sub-lethal concentrations of malathion. There was a marked inhibition in DNA, RNA and protein contents and the activities of acid and alkaline phosphatase. ...
... Pesticides play an important role in maintaining agricultural production through the protection of all types of crops from pest attack and vector-borne diseases, but some pesticides have adverse effects on fish and other non-target animals. Organophosphate (OP) compounds are extensively used in the fields of agriculture as well as public health, making up 50% of the insecticides use world wide, because such compounds can degrade promptly due to their non-persistent nature [5]. Wacksman et al. pointed out that OP could inhibite the activity of acetylcholinesterase (AChE), and consequently, acetylcholine accumulated in the cholinergic receptors of the central and peripheral nervous systems [6]. ...
... Different behavioral changes mostly were noticed as less activity, lost of equilibrium, restlessness, abnormal swimming behavior, rapid gill movement, staying motionless on the aquarium bottom and change in the color of zebrafish skin which are similar to the observations of behavioural responses reported in previous studies [17]- [19]. It was reported that these behavioral abnormalities of fish (Labeo rohita) were observed only at high concentrations (values higher than 96 h LC 50 ) and after exposure to various acute concentrations (5,10,15,20,25 and 30 mg· L -1 ) of MAL [20]. Enzymatic changes was considered to be important indication of hazardous chemicals. ...
... sometimes conflicting (Sprague, 1985; Chapman, 1995). Solubilization of lipophilic xenobiotics in acetone prior to experiments (Toledo and Jonsson, 1992; Biagianti-Risbourg et al., 1996) or during the tests (Bailey and Liu, 1980; Kumar and Ansari, 1986; Ramalingan, 1988; Mishra and Shukla, 1994) may alter the toxic effects of chemicals on fish although the low concentrations of acetone used (0.02 to 0.1%) are not acutely toxic to fish (Bailey and Liu, 1980; Biagianti-Risbourg et al, 1996). We investigated the effects of acetone on the hepatocytic responses of Oncorhynchus mykiss sac-fry by electron microscopy as, to our knowledge, no information is available on this subject. ...
... Acetone has been used as vehicle in some toxicological tests using fish to solubilize the liposoluble chemicals tested (Kumar and Ansari, 1986; Ramalingan, 1988; Mishra and Shukla, 1994). Acetone changes the absorption of xenobiotic molecules in Rode&a and increases their hepatotoxicity (Raymond and Plaa, 1995; Heylings et al., 1996). ...
Acetone has been used to solubilize liposoluble molecules in toxicological tests using fish. The effects over time of a sublethal concentration (0.1%) of acetone on the livers of trout sac-fry (Oncorhynchus mykiss) were investigated by electron microscopy. Changes in the hepatocyte membranes (dilation of the interhepatocytic space, formation of intracellular myelinic figures) occurred between the first and 7th day of exposure. However, these changes were minor and they did not persist after 22 days of exposure. Acetone caused changes in hepatocytes which could affected the toxicity of the xenobiotics tested. Thus use of this solvent as vehicle in toxicological experiments should be avoided.
... RNA has a direct relationship with protein content and an indirect relationship with DNA content. The reduced content of RNA in the present study was due to proteolysis and destruction of protein synthesis or inhibiting the RNA synthesis process Mukherjee, 2000, 2003;Kumar and Ansari, 1986). Thus, malathion inhibits RNA synthesis, which results in the depletion of RNA and subsequent protein content, as observed in the present study. ...
... For example: in the liver of Channa punctatus exposed to cythion (Narain and Sathyanesan, 1985), Oreochromis mossambicus exposed to endosulfan (Ganesan et al., 1989), and in Gambusia offinis exposed to tannery effluent (Revathi et al., 2005); in the blood of Catla catla exposed to mercury (Rai, 1987), and in Mystus vittatus exposed to metasystox and sevin (John. 2007); in the ovary of Brachydanio rerio exposed to malathion (Ansari and Kumar, 1987), and in Gambusia offinis exposed to tannery effluent (Revathi et al., 2005); in both testis and ovary of C.fasciata exposed to arsenic (Shukla and Pandey, 1986). The aminoacid concentration in acute sublethal exposure of test effluent to M. vittatus was increased significantly and found significant at p<0.05 and 0.001 (Fig.1). ...
The effects of whole Rayana paper board industries limited effluent (W.R.P.B.I.L.E) on some biochemical aspects of the fresh water teleost fish, Mystus vitatus were studied under experimental conditions through the course of the annual reproductive cycle of these fishes. A quantitative estimation of the amino acids and protein content was made in liver tissues, blood and gonadial tissues. The content of all these nutrient and building materials was observed to be significantly higher during the spawning phase when compaired to the other phases of the annual reproductive cycle. The changes produced in these biochemical parameters on account of exposure of the fishes for 96-h to 0.4 and 0.8 of LC50 – 96-h of, effluent, were also studied during all the three phases of the annual reproductive cycle. All the stress – induced biochemical alterations were always found to be the greater in case of 0.8 than in case of 0.4 e. effluent both the sublethal concentrations of the annual reproductive cycle of the fish, the aminoacid content of liver, blood and gonads was found to be increased, while the protein, content was measured to be decreased in liver, blood and gonads. The present study concludes a stress-induced metabolic dysfunction in response to effluent toxicity in the fish.
... Fungal cutinase also presents higher efficiency than yeast esterase in degrading malathion [47], an organophosphate insecticide and acaricide [48] which has severe toxic effects [49][50][51]. Another advantage of cutinase is that it produces less toxic end products than esterase in malathion degradation [47]. ...
Cutinases (EC 3.1.1.74) are widely distributed in fungi, bacteria and plants with diversified structures and properties. Besides acting on the natural substrate cutin, cutinases are the first line of natural biocatalysts to hydrolyze artificial polyesters and toxic xenobiotics such as polyethylene terephthalate (PET), polycaprolactone (PCL), polylactic acid (PLA), polyhydroxybutyl succinate (PBS), phthalate and malathion esters. Moreover, cutinases can act as promising stereoselective catalysts in esterification and transesterification reactions and present better selectivities than lipases. These pioneering studies indicate that the biotechnological application of cutinase as a powerful tool in synthetic biology deserves further investigation, for both degradation and biosynthesis towards a broader range of ester bond-containing substrates. This review summarizes the classifications and properties of cutinases from different sources and insights into the structure–function relationship of different cutinases. It also highlights the uniqueness and advantages of representative cutinases in biodegradation and biosynthesis, and then prospects the future application of natural and engineered cutinases in synthetic biology.
... 3). As like acetylcholinesterase, the acid phosphatase has serine residue at its active site and the organophosphorus pesticides are generally inhibitors of serine containing enzymes (Cao et al., 2020;Dyk et al., 2011;Kumar et al., 1986). With the addition of organophosphorus pesticides, the enzymatic activity of ACP is inhibited and produces less ascorbic acid, which results in the increased catalytic oxidation of TMB over Fe-N/C SAzyme oxidase mimetics. ...
In this study, a novel highly sensitive colorimetric platform has been designed for malathion assay based on Fe-N/C SAzyme. The as-synthesized SAzyme can directly oxidize 3,3´,5,5´-tetramethylbenzidine (TMB) to generate blue colored oxidized TMB. L-ascorbic acid-2-phosphate (AA2P), a substrate of acid phosphatase (ACP), could be hydrolyzed to AA, thereafter inhibit the oxidization reaction of TMB, leading to a conspicuous blue color fading. With the addition of malathion hindered the ACP activity and limited the AA production, resulting in the recovery of the catalytic activity of single-atom nanozyme. Under optimized operational conditions, a novel colorimetric assay has been designed for malathion detection with LOD of 0.42 nM. Besides, quantification of malathion in environmental and food samples was achieved based on the proposed strategy. In addition, the successfully integrated paper/smartphone sensor provided sensitive, and rapid, reliable detection of malathion with a LOD of 1 nM.
... Malathion has adverse effects, for instance, it is a cholinesterase inhibitor (Chandra 2008), deactivating the enzyme leading to production of acetylcholine in synapse. It decreases DNA, RNA and protein contents (Kumar K and Ansari 1986), declines antioxidant defense (Patil and David 2013) and makes histological changes in organs (Richmonds and Dutta A c c e p t e d M a n u s c r i p t 1992). Pesticide mixture is common in the aquatic ecosystems including rivers, streams and lakes (Gilliom 2007). ...
DNA breakage has been frequently used as a biomarker of the pesticide toxicity. The present study introduced a method to quantify the DNA breakage in Oncorhynchus mykiss exposed to the pesticide Malathion. Specimens were exposed to different concentrations of malathion for 1-9 days and their gill and liver were sampled. DNA was extracted and electrophoresed using agarose gel. The pixel density curves were obtained from the gel smears. The area under the curves were arbitrarily divided from 3 up to 7 segments using a Java macro in the software ImageJ. Some weighted averaging methods were used to calculate DNA breakage in each gel lane. Akike information Criteria (AIC) was used to find the best analysis of variance (ANOVA). The liver was more sensitive than the gill showing a larger number of significant differences among the specimens exposed to various concentrations of malathion. The geometric weighted averaging on the data extracted from the 7-segment pixel density curve resulted to the lowest AIC. The double-strand DNA breakage of O. mykiss was able to detect malathion in freshwater in concentrations over 0.05 mg L⁻¹.
... In fish, exposure to pollution, for short and long periods and also depending on stage of ovarian and/or follicular development, may result in reduction in fertility leading to decreases in the gonadosomatic index (GSI), smaller and less developed oocytes, smaller number of mature oocytes and increases in the quantity of atretic follicles (Kime, 1995). Decreases in nucleic acids, protein content and enzymatic activity by alkaline phosphatase after exposure to malathion, an organophosphate insecticide, were described by Ansari and Kumar (1987). Exposure to heavy metals, cadmium and mercury was also reported causing decreases in fish fertility (Ram and Sathyanesan 1986;Victor et al., 1986;Ram and Sathyanesan, 1987;El-Ebiary et al., 2013). ...
Studies on reproduction in sea turtles are important due to its life cycle, migratory patterns, high juvenile mortality and environmental impacts. This study aimed to analyse histomorphometrically gonads of C. mydas from the coastline of the Espírito Santo State, Brazil. Ovaries and testicles were collected between 2014 and 2015 from stranded animals. The material was fixed in formalin 10%, assessed macroscopically and processed for histomorphometrical evaluation. Gonads from 34 individuals were evaluated, twenty-four females and ten males. Macroscopic sexual identification presented 100% accuracy confirmed by histology. Sexual dimorphism was observed in one individual, which was considered as adult (CCL=1.023 m). Microscopy of female gonads revealed predominant previtellogenic follicles; oocyte diameter ranged between 161µm and 750µm and a positive correlation between ovarian length, largest oocyte and CCL was found. In males, autolysis was verified in five individuals. Viable testicles revealed predominant spermatogonia, primary spermatocytes and Sertoli cells in the seminiferous tubules and, Leydig cells and fibroblasts in the stroma. There was a positive correlation between tubular diameter and CCL and testicle length and CCL. Maturation of stromal tissue and a positive correlation between tubular lumen and CCL were also observed. Gonad development is proportional to individual growth.
... More interestingly, many of adult fish toxicity studies can be replaced by the zebrafish embryos [22]. The adult zebrafish was used in many toxiological measurements such as the testing of uranium and lead [23], anilines [24], malathion [25], and metronidazole [26]. It is noteworthy to mention that the toxicity of the nanoparticles can be effectively tested using the zebrafish embryos [21]. ...
The recent application of 2D MXene nanomaterials as adsorbents and membranes for water treatment as well as biomedical applications is attracting a growing interest. However, the environmental impact of Ti3C2Tx (MXene), especially their potential risks on aquatic biota and ecosystems of aquatic environment has never been explored. Herein, we have studied the biocompatibility of Ti3C2Tx by analyzing their potential toxicity in vivo using the zebrafish embryo model. Ti3C2Tx morphology, surface charge, and stability were characterized by SEM, TEM, x-ray diffraction spectroscopy. The aggregation pattern of Ti3C2Tx suspension in seawater was investigated. The ICP-MS results showed that the zebrafish embryos can uptake Ti3C2Tx in dose dependent manner. Acute toxicity of the attached/internalized Ti3C2Tx was tested at concentrations of 25, 50, 100 and 200 µg/mL. According to the 96-hours sigmoidal mortality curve, the LC50 of Ti3C2TX was calculated to be at 257.46 μg/mL and the highest NOEC (< 20% mortality) was at 50 µg/mL. The LOEC (≥20% mortality) of Ti3C2Tx was detected at 100 µg/mL, as this concentration showed slight increase in mortality (21%). However, no significant teratogenic effects were observed on zebrafish embryos at 100 µg/mL. This nontoxicity was confirmed by the locomotion and the neurotoxicity assays, as 50 µg/mL of Ti3C2TX showed no harmful effect on the neuromuscular activities. In conclusion, because the LC50 of Ti3C2Tx was greater than 100 µg/mL, it can be classified within the “practically nontoxic” group according to the Acute Toxicity Rating Scale by Fish and Wildlife Service (FWS), thus we suggest the safe uses and discharge of Ti3C2Tx MXene in the aquatic ecosystem at concentrations below 100 µg/mL.
... 93,94 A group led by Prof. Badre Alam Ansari at Kashi Naresh Government Postgraduate College (KNGPGC), Gyanpur had research to understand the toxicity of Diazinon and Malathion on zebrafish's liver, ovary, brain, and skeletal parts. [95][96][97][98][99][100] Dr. Uma's group at Tamil Nadu Fisheries University (TNFU), Chennai proved that zebrafish is an alternative model for assessing the environmental chemical toxicity in the laboratory condition. They checked the toxic effect of Ivermectin and Cypermethrin on the embryos, indicating that low levels of these drug contaminations in the aquatic environment could affect the developmental stages of aquatic organisms. ...
The zebrafish (Danio rerio) is a versatile model organism that has been used in biomedical research for several decades to study a wide range of biological phenomena. There are many technical advantages of using zebrafish over other vertebrate models. They are readily available, hardy, easy, and inexpensive to maintain in the laboratory, have a short life cycle, and have excellent fecundity. Due to its optical clarity and reproducible capabilities, it has become one of the predominant models of human genetic diseases. Zebrafish research has made rapid strides in the United States and Europe, but in India the field is at an early stage and many researchers still remain unaware of the full research potential of this tiny fish. The zebrafish model system was introduced into India in the early 2000s. Up to now, more than 200 scientific referred articles have been published by Indian researchers. This review gives an overview of the current state of knowledge for zebrafish research in India, with the aim of promoting wider utilization of zebrafish for high level biological studies.
... The zebrafish has been extensively used in acute toxicological studies [reviewed by [12,16,5,17]]. Examples include the use of adult zebrafish for the testing of lead and uranium [18], malathion [19], colchicine [20], anilines [21], and metronidazole [22]; and the use of juveniles for testing agricultural biocides [23]. Zebrafish embryos are also being used in toxicity studies (reviewed by [24]). ...
Cannabinoids are natural or synthetic compounds related chemically to Δ -THC, the principle psychotropic constituent of
the hemp plant, Cannabis sativa L. Here, we examine the effects of the cannabinoids Δ -THC, WIN 55,212–2 and CP
55,940, and the cannabinoid antagonist (Am 251). Exposures were either acute (1–12h exposure at 108 hour post
fertilization [hpf]); or chronic (96h exposure starting at 24 hpf. Wild type zebrafish embryos (3,250, including controls),
were cultured individually in 250μl defined buffer in 96-well plates. Geometric range-finding was used to determine the
experimental concentrations At day 5, behavioural analysis (visual motor response test) was carried out in which
movement of individual larvae was analysed using automated video-tracking. With acute exposure, embryos showed a
biphasic response to the dark challenge with Δ -THC, WIN55, 212–2 and CP55,940. This response consisted of stimulation
of locomotor activity at low concentrations, suppression at high doses. With the antagonist AM251 alone, the locomotor
activity was suppressed at high concentrations. With chronic exposure, embryos habituated to the effects of all three
cannabinoids when assayed with the dark challenge phase. Furthermore, the excitation was ameliorated when antagonist
was co-administered with the cannabinoid. We conclude that cannabinoids have similar effects in zebrafish and mammals.
In particular, the acute exposure response resembles behavioural effects reported for adult rodents.
... 4,[10][11][12][13][14][15][16][17] The zebrafish has been extensively used in toxicological studies to screen either single compounds or small panels of compounds (reviewed by refs. [18][19][20][21][22]. Examples include the use of adult zebrafish for the testing of lead and uranium, 23 malathion, 24 colchicine, 25 anilines, 26 and metronidazole, 27 and the use of juveniles and embryos for testing agricultural biocides. 28,29 Not only adult zebrafish, but also zebrafish embryos or larvae are used in toxicity studies (reviewed by Truong et al. 30 ). ...
Abstract The zebrafish larva is a promising whole-animal model for safety pharmacology, environmental risk assessment, and developmental toxicity. This model has been used for the high-throughput toxicity screening of various compounds. Our aim here is to identify possible phenotypic markers of teratogenicity in zebrafish embryos that could be used for the assaying compounds for reproductive toxicity. We have screened a panel of 60 water-soluble toxicants to examine their effects on zebrafish development. A total of 22,080 wild-type zebrafish larvae were raised in 250 μL defined buffer in 96-well plates at a plating density of one embryo per well. They were exposed for a 96-h period starting at 24 h post-fertilization. A logarithmic concentration series was used for range-finding, followed by a narrower geometric series for developmental toxicity assessment. A total of 9017 survivors were analyzed at 5 days post-fertilization for nine phenotypes, namely, (1) normal, (2) pericardial oedema, (3) yolk sac oedema, (4) melanophores dispersed, (5) bent tail tip, (6) bent body axis, (7) abnormal Meckel's cartilage, (8) abnormal branchial arches, and (9) uninflated swim bladder. For each toxicant, the EC50 (concentration required to produce one or more of these abnormalities in 50% of embryos) was also calculated. For the majority of toxicants (55/60) there was, at the population level, a statistically significant, concentration-dependent increase in the incidence of abnormal phenotypes among survivors. The commonest abnormalities were pericardial oedema, yolk sac oedema, dispersed melanophores, and uninflated swim bladder. It is possible therefore that these could prove to be general indicators of reproductive toxicity in the zebrafish embryo assay.
... Little information has been published on macroscopic changes in the colour of fish livers. However, laboratory tests and field studies have demonstrated that fish exposed to pesticides show pathologic changes in the liver (Kumar & Ansari 1986, Gill et al. 1988) and that OPs are reported to have negative effects on the antioxidant system of carp and therefore lead to 'oxidative stress' (Hai et al. 1997). This 'oxidative stress' has been found to be the cause of jaundice, which causes yellowish decolouration in the liver of yellowtails Seriola quinqueradiata (Sakai et al. 1998). ...
Effects of dissolved pesticides on fish are widely described, but little is known about effects of pesticide-contaminated feeds taken up orally by fish. In integrated farms, pesticides used on crops may affect grass carp that feed on plants from these fields. In northern Vietnam, grass carp suffer seasonal mass mortalities which may be caused by pesticide-contaminated plants. To test effects of pesticide-contaminated feeds on health and bioaccumulation in grass carp, a net-cage trial was conducted with 5 differently contaminated grasses. Grass was spiked with 2 levels of trichlorfon/fenitrothion and fenobucarb. Unspiked grass was used as a control. Fish were fed at a daily rate of 20% of body mass for 10 d. The concentrations of fenitrothion and fenobucarb in pond water increased over time. Effects on fish mortality were not found. Fenobucarb in feed showed the strongest effects on fish by lowering feed uptake, deforming the liver, increasing blood glucose and reducing cholinesterase activity in blood serum, depending on feed uptake. Fenobucarb showed increased levels in flesh in all treatments, suggesting bio-concentration. Trichlorfon and fenitrothion did not significantly affect feed uptake but showed concentration-dependent reduction of cholinesterase activity and liver changes. Fenitrothion showed bioaccumulation in flesh which was dependant on feed uptake, whereas trichlorfon was only detected in very low concentrations in all treatments. Pesticide levels were all detected below the maximum residue levels in food. The pesticide-contaminated feeds tested did not cause mortality in grass carp but were associated with negative physiological responses and may increase susceptibility to diseases.
... Similar to present study, Mazorra et al. (2002) also observed significant inhibition in ALP activity in digestive gland and in gill when exposed to mercury. A dose-dependent inhibition of alkaline phosphatase activity was reported in the liver of zebra fish (Brachydanio rerio) exposed to Malathion (Kumar and Ansari 1986). Sharma (1990) also showed consistent inhibition of ALP activity in liver, kidney and muscle of Channa gachua exposed to endosulfan for a longer period (15-30 days). ...
Edible oyster, Crassostrea rivularis were collected from North Wandoor (NW) and Phoenix Bay Jetty (PJ) of Andaman and their biological, biochemical and nutritional status were assessed. Water quality parameters were estimated from both places and recorded higher levels of nitrite, nitrate and phosphate in PJ samples compared to NW. There was significant difference in length and weight of oysters collected from the 2 sites. Oysters from NW samples were bigger in size compared to PJ samples. All biochemical parameters showed reduction in the C. rivularis collected from PJ compared to NW. The extent of reduction in liver glycogen and ascorbic acid, acetyl choline esterase and alkaline phosphatase activities in gill tissue were 65, 42, 17.07 and 46.01% respectively. Overall results suggested that, oysters dwelling at PJ were not healthy and under stress due to continuous exposure to organic discharge which in turn might have caused lower growth rate of the oyster and depression in the biochemical and enzyme activities compared to NW oysters
... However, the presence of chlorine inhibited ALP activity in all the organs tested, which might be an indication of role of chlorine in inhibiting protein synthesis. Similarly, a dose dependent inhibition of ALP was reported in liver of Zebra danios exposed to malathion [46]. A consistent inhibition of ALP activity in liver, kidney and muscle was reported when Channa gachua was exposed to endosulfan, an organochlorine pesticide [47]. ...
Thermal effluents discharged through cooling systems of nuclear power plants often contain chlorine (used to control bio-fouling), which may affect the metabolic status of fishes. In order to evaluate the hypothesis, we tested the effect of high temperature and a persistent sub-lethal chlorine exposure on stress responses in Cyprinus carpio advanced fingerlings. Fishes were acclimated to four different temperatures (26, 31, 33, and 36°C) and maintained for 30 days in two different groups. Subsequently, one of the groups was exposed to persistent chlorine (0.1mgL−1) for another 28 days and was compared with their respective temperature controls (without chlorine exposure). Sub-lethal doses of pollutants and increasing temperatures with in the tolerance range may not always register any morphological changes Therefore, we studied organ specific biochemical pathways viz. aspartate amino transferase, alanine amino transferase (enzymes of protein metabolism) in liver and muscle; fructose 1,6 diphosphatase (gluconeogenic pathway), in liver; pyruvate kinase, malate dehydrogenase, and lactate dehydrogenase (glycolytic pathway) in muscle; glucose-6-phosphate dehydrogenase (pentose phosphate pathway) in liver; alkaline phosphatase (phosphorus metabolism) in intestine, liver, and muscle; acetylcholine esterase (neurotransmitting enzyme) in brain, and adenosine triphosphate (for membrane transport) in gills at two different acclimation periods (14 and 28 days). The results indicate that C. carpio fingerlings demonstrated metabolic readjustments with increasing temperatures, in order to cope with energy demand of the cell. However, exposure to chlorine at higher temperatures affected protein metabolism, gluconeogenic pathway and subsequently glycolytic pathway, leading to an energy-limited condition. In addition, alteration of membrane transport and neurotransmission might be an early indication of cellular damage. Overall results indicate that persistent sub-lethal chlorine exposure elicits temperature induced stress response in C. carpio early fingerlings.
... The oogenesis of the was protein level at 0 day (8.5 ± 0.4mg/g) in the muscle. in the late perinucleolar stage and in the beginning of Kumar and Asari, [11] concluded that the marked th inhibition DNA, RNA, Protein, Acid and Alkaline metabolic routes by which an organism can detoxify phosphatase in the liver of four month old Zebra Fish organophosphorus insecticide. In addition, the after 7 day of exposure of different concentration of physiological condition of the organism during toxic th malathion (0.5,0.7,0.9 and 1.1 mg/l). ...
4 Abstract: The present study deals with the effect of some water pollutant; malathion as an insecticide on histological and histochemical study in ophicoephalus punctatus. The exposure of fish to the examined pollutants induced a disappearance of some fractions and consequently changes of relative mobility and area that indicate damage. Ophicoephalus punctatus was exposed for 7 days to Malathion at a concentration of 10, 12, 14,16,18,20 µ1/1 and 100ppm under laboratory conditions. The fish showed severe histological changes in brain, liver, ovary and tissues. The degenerative changes included hypertrophy of cells and their nuclei, liver on the whole showed distance appearance. Pycnosis, Vaculation, Necrosis, fragmentation of ova were recorded in an increasing order towards the higher tested doses.
... Acute toxicity studies using zebrafish are limited. Examples of toxicants investigated include lead and uranium (Labrot et al., 1999), malathion (Kumar and Ansari, 1986), metronidazole (Lanzky and Halling-Sorensen, 1997), anilines (Zok et al., 1991), and colchicines (Roche et al., 1994). ...
Zebrafish have been used predominantly in developmental biology and molecular genetics, but their value in toxicology as well as drug discovery has been recognized. To evaluate the toxicity of a chemical, it is essential to identify the endpoints of toxicity and their dose-response relationships, elucidate the mechanisms of toxicity, and determine the toxicodynamics of the chemical. In addition to detailed toxicological investigations of a single chemical, there also is a need for high-throughput large- scale screening for toxicity of several hundreds of chemicals at a time. In both cases, the zebrafish has numerous attributes. More is probably known about ''what is normal'' in the zebrafish than any other fish species. This includes morpho- logical, biochemical, and physiological information at all stages of early development and in juveniles and adults of both sexes. This makes using the zebrafish ideal for toxicology research where the objective is to identify adverse effects of chemical exposure.
... Similar to present study, Mazorra et al. (2002) also observed significant inhibition in ALP activity in digestive gland and in gill when exposed to mercury. A dose-dependent inhibition of alkaline phosphatase activity was reported in the liver of zebra fish (Brachydanio rerio) exposed to Malathion (Kumar and Ansari 1986). Sharma (1990) also showed consistent inhibition of ALP activity in liver, kidney and muscle of Channa gachua exposed to endosulfan for a longer period (15–30 days). ...
Oysters filter up to 1,500 litres of sea water per day looking for food. This may lead to >100 times higher concentrations of pollutants in the oysters than surrounding waters. Oysters have wide geographical distribution. They are sensitive to environmental pollutants and accumulate anthropogenically derived chemicals at a high rate (Roesijadi 1994a, 1994b, Binelli et al. 2006). Biochemical and molecular responses of fishes to aquatic stressors are the earliest indicators of environmental perturbations to animal which can be useful as early signs of organismal alteration (Pal and Ayyappan 2002). Biochemical alteration of oysters and mussels are widely used in pollution monitoring studies (Cajaraville et al. 2000, Lau and Wong 2003, Roméo et al. 2003, Silva et al. 2003, Zanette et al. 2006, Bernal-Hernández et al. 2010). Many biomarkers are extensively used globally for pollution monitoring (Livingstone 1993, Kaaya et al. 1999). Biochemical parameters are extensively used to characterize anthropogenic pollution (Burgeot et al. 1996, Cajaraville et al. 2000, Zanette et al. 2006).
... All effects that were observed in either the gills or the intestine and in the liver induce behavioral changes as a consequence of the decrease in the general state of health of C. paleatus. This will, together with the inhibition of plasma cholinesterase (Silva et al., 1993) and the effect on gonads (Ansari and Kumar, 1987b), affect the chances of survival of these fish in the natural environment. They will have difficulties in detecting, identifying, and responding in an appropriate way to natural chemical stimulation, which is an important component of environmental physiology (Post and Leasure, 1974;Lemly and Smith, 1986;Silva et al., 1993). ...
The effects of contamination, through water or food, of a sublethal dose of the organophosphate methyl parathion were analyzed in tissues that are responsible for absorption (gills, intestine) and metabolism (liver), in the freshwater fish Corydoras paleatus. In gill respiratory lamellae, epithelial hyperplasia, edema, and detachment occurred, diminishing sooner after contamination by food than after contamination through water. In the intestine, lipoid vacuolization of enterocytes, apical cytoplasm, and an increase in goblet cell activity occurred mainly after ingestion of contaminated food. The liver exhibited cloudy swelling, bile stagnation, focal necrosis, atrophy, and vacuolization after contamination through both absorption routes, the highest degeneration being between T8 and T24. Metabolic processes that depend on liver function were equally impaired by the two routes of contamination, but secondary effects vary with gill and intestine pathologies as a consequence of water and food contamination, respectively. Therefore, a “safe” sublethal dose of methyl parathion causes serious health problems in C. paleatus.
... The zebrafish has been extensively used in acute toxicological studies [reviewed by [12,16,5,17]]. Examples include the use of adult zebrafish for the testing of lead and uranium [18], malathion [19], colchicine [20], anilines [21], and metronidazole [22]; and the use of juveniles for testing agricultural biocides [23]. Zebrafish embryos are also being used in toxicity studies (reviewed by [24]). ...
The zebrafish is a powerful whole animal model which is complementary to in vitro and mammalian models. It has been shown to be applicable to the high-throughput behavioral screening of compound libraries. We have analysed 60 water-soluble toxic compounds covering a range of common drugs, toxins and chemicals, and representing various pharmacological mechanisms. Wild-type zebrafish larvae were cultured individually in defined buffer in 96 well plates. They were exposed for a 96h period starting at 24h post fertilization (hpf). A logarithmic concentration series was used for range-finding, followed by a narrower geometric series for LC(50) determination. LC(50) values were determined at 24h intervals and behavioral testing was carried out on day 5. We used the visual motor response test, in which movement of individual larvae was analysed using automated video-tracking. For all compounds, LC(50) values were found to decrease as the embryo developed. The majority of compounds (57/60) produced an effect in both the basal (lights on) and challenge phases (lights off) of the behavioral assay. These effects were either (i) suppression of locomotor activity (monotonic concentration-response); (ii) stimulation then suppression (biphasic response); (iii) stimulation (monotonic response). We conclude that behavioral assays with zebrafish embryos could be useful for pharmaceutical efficacy and toxicity screening. The precise phenotypic outcome obtained with behavioral assay varies with compound class.
... The zebrafish is increasing being used in toxicological studies [reviewed by: 13,14]. Example include the use of adult zebrafish for the testing of lead and uranium [15], malathion [16], colchicine [17], anilines [18], and metronidazole [19]; and the use of juveniles for testing agricultural biocides [20]. Zebrafish embryos are also being used in toxicity studies [reviewed by: 21]. ...
In the drug discovery pipeline, safety pharmacology is a major issue. The zebrafish has been proposed as a model that can bridge the gap in this field between cell assays (which are cost-effective, but low in data content) and rodent assays (which are high in data content, but less cost-efficient). However, zebrafish assays are only likely to be useful if they can be shown to have high predictive power. We examined this issue by assaying 60 water-soluble compounds representing a range of chemical classes and toxicological mechanisms.
Over 20,000 wild-type zebrafish embryos (including controls) were cultured individually in defined buffer in 96-well plates. Embryos were exposed for a 96 hour period starting at 24 hours post fertilization. A logarithmic concentration series was used for range-finding, followed by a narrower geometric series for LC(50) determination. Zebrafish embryo LC(50) (log mmol/L), and published data on rodent LD(50) (log mmol/kg), were found to be strongly correlated (using Kendall's rank correlation tau and Pearson's product-moment correlation). The slope of the regression line for the full set of compounds was 0.73403. However, we found that the slope was strongly influenced by compound class. Thus, while most compounds had a similar toxicity level in both species, some compounds were markedly more toxic in zebrafish than in rodents, or vice versa.
For the substances examined here, in aggregate, the zebrafish embryo model has good predictivity for toxicity in rodents. However, the correlation between zebrafish and rodent toxicity varies considerably between individual compounds and compound class. We discuss the strengths and limitations of the zebrafish model in light of these findings.
... In the context of toxicity, the zebrafish finds application in drug safety testing and ecotoxicological screening. For further examples, see Table 3. Chronic exposure regimes have been used to assess the toxicity of lead and uranium (Labrot et al., 1999), colchicine (Roche et al., 1994), anilines (Zok et al., 1991), metronidazole (Lanzky and Halling-Sorensen, 1997), and agricultural biocides (Kumar and Ansari, 1986;Gorge and Nagel, 1990). Acute toxicity studies are fewer. ...
Technological innovation has helped the zebrafish embryo gain ground as a disease model and an assay system for drug screening. Here, we review the use of zebrafish embryos and early larvae in applied biomedical research, using selected cases. We look at the use of zebrafish embryos as disease models, taking fetal alcohol syndrome and tuberculosis as examples. We discuss advances in imaging, in culture techniques (including microfluidics), and in drug delivery (including new techniques for the robotic injection of compounds into the egg). The use of zebrafish embryos in early stages of drug safety-screening is discussed. So too are the new behavioral assays that are being adapted from rodent research for use in zebrafish embryos, and which may become relevant in validating the effects of neuroactive compounds such as anxiolytics and antidepressants. Readouts, such as morphological screening and cardiac function, are examined. There are several drawbacks in the zebrafish model. One is its very rapid development, which means that screening with zebrafish is analogous to "screening on a run-away train." Therefore, we argue that zebrafish embryos need to be precisely staged when used in acute assays, so as to ensure a consistent window of developmental exposure. We believe that zebrafish embryo screens can be used in the pre-regulatory phases of drug development, although more validation studies are needed to overcome industry scepticism. Finally, the zebrafish poses no challenge to the position of rodent models: it is complementary to them, especially in early stages of drug research.
... Acute toxicity studies using zebrafish are limited. Examples of toxicants investigated include lead and uranium ( Labrot et al. 1999), malathion (Kumar and Ansari, 1986), metronidazole (Lanzky and Halling-Sorensen, 1997), anilines (Zok et al., 1991), and colchicines (Roche et al., 1994). ...
Zebrafish (Danio rerio) has been a prominent model vertebrate in a variety of biological disciplines. Substantial information gathered from developmental and genetic research, together with near-completion of the zebrafish genome project, has placed zebrafish in an attractive position for use as a toxicological model. Although still in its infancy, there is a clear potential for zebrafish to provide valuable new insights into chemical toxicity, drug discovery, and human disease using recent advances in forward and reverse genetic techniques coupled with large-scale, high-throughput screening. Here we present an overview of the rapidly increasing use of zebrafish in toxicology. Advantages of the zebrafish both in identifying endpoints of toxicity and in elucidating mechanisms of toxicity are highlighted.
SUMMARY
The present study aimed to evaluate the relative toxicological safety of certain non-conventional insecticides, i.e. imidacloprid, neem oil, mineral oil and a commercial product of the entomopathogenic fungi, Beauvaria bassiana (all compared to chlorpyrifos and cypermethrin). In this respect, acute and subchronic studies on Japanese quail, Coturnix coturnix japonica and Bolti fish, Tilapia nilotica were conducted. For subchronic studies, sublethal daily doses (or concentrations) each of 1/50 or 1/100 LD50 (or LC50) were used for 30 days.
1. Acute toxicity studies:
Against Japanese quail birds, the insecticides could be comparatively classified as follows: chlorpyrifos and imidacloprid are extremely toxic (LD50’s: 8.8 and 25 mg/kg b.w, respectively); cypermethrin moderately toxic (LD50’s 1247 mg/kg); neem oil and mineral oil, relatively non-toxic (LD50’s: > 10000 and 20000 mg/kg, respectively).
Dermally, chlorpyrifos is classified as very toxic (LD50: 155.78 mg/kg), imidacloprid as moderately toxic (LD50: 1757.9 mg/kg), cypermethrin, neem or mineral oil as relatively non-toxic (LD50’s: 8957.4, > 50000 mg/kg, respectively). The entomopathogenic fungi showed low acute oral or dermal toxicity to quail (LD¬50’s: 6 x 108 and > 30 x 108 conidia/kg b.w. equivalent to 20 and 100 ml of the liquid formulation/kg b.w., respectively).
Against fingerlings of Bolti fish and using static methods, the tested insecticides could be classified as follows: chlorpyrifos and chypermethrin, extremely toxic (LC50’s: 0.06 and 0.028 ppm, respectively; imidacloprid moderately toxic (LC50: 13 ppm); neem oil and mineral oil, relatively non-toxic (LC50’s: 150 and 120 ppm, respectively).
2. Subchronic toxicities:
2.1. Against quail birds:
Quail birds were treated orally with single daily doses (each of 1/50 or 1/100 LD50) for 30 days. LD50’s of neem and mineral oils were supposed to be 10000 and 20000 mg/kg b.w, respectively. No clinical signs of toxicity could be observed throughout the experimental period. Birds treated with mineral oil or cypermethrin showed no deaths within test period. Birds treated with 1/50 LD50 of chorpyrifos, neem oil or the entomopathogenic fungi showed 13.3, 20 and 33.3% mortalities, respectively. At 1/100 LD50 the toxicity was less powerful (mortalities were nearly 6.6%). Except for the entomopathogenic fungi, all the tested toxicants caused significant reduction in red blood cells count, Hemoglobin content and hematocrit values, whereas white blood cells were increased. The effects were less powerful for the dose 1/100 LD50. Chlorpyrifos was the only insecticide that significantly decreased the specific activity of AChE which was determined in brains of birds. Chlorpyrifos, imidacloprid, neem oil and the fungal product significantly reduced the activity of non-specific esterases at 1/50 LD50 but were of no effect at 1/100 LD50. Cypermethrin and mineral oil exhibited no effect on esterase activity at the two tested doses.
The mineral oil and the fungal product caused no significant alterations in the activities of alkaline phosphatase, alanine amino transferase; total protein, bilirubin concentration and albumin at the two tested doses. Chlorpyrifos, cypermethrin, imidacloprid and neem oil significantly elevated alkaline phosphatase, transaminases, total bilibrubin, albumin whereas reduced cholesterol and total lipids. The effect was more remarkable for the conventional insecticides, chlorpyrifos and cypermethrin. It could be concluded that alterations caused in the tested biochemical measurements due to chlorpyrifos, cypermethrin, imidacloprid and neem oil might indicate the liver dysfunction. The effect was more pronounced in chlorpyrifos and cypermethrin and generally less powerful at the dose 1/100 LD50. This was emphasized by the histopathological examination where livers of birds treated with mineral oil or the entomopathogenic fungi showed no histological changes and seemed to be normal. In contrary, livers of birds treated with chlorpyrifos, cypermethrin, imidacloprid and neem oil showed similar histopathological reactions differ in severity and generally appeared as vacoular degeneration of hepatocytes, necrosis, infiltration, congestion and others.
Concerning the effect on kidney function, it was found that the mineral oil and the fungal product exhibited no significant effect on urea and creatinine concentrations. Chlorpyrifos, cypermethrin, imidacloprid and neem oil showed significant increase of these parameters. It is concluded that chlorpyrifos might be severely nephrotoxic while cypermethrin, imidacloprid and neem oil might be less powerfull. The entomopathogenic product and the tested mineral oil might be relatively of no-renal toxicity.
For the effect on blood sugar content results showed that only the tested conventional insecticides (i.e. chlorpyrifos and cypermethrin) were found to elevate glucose concentration at the two tested doses.
2.2. Bolti fish, Tilapia nilotica:
Bolti fish fingerlings were continuously exposed to freshly prepared concentrations (1/50 or 1/100 LC50) of the insecticides for 30 days. Few number of fish died throughout the test period (3.33-10%). The highest mortalities were observed for neem and mineral oils (10%). At the concentration, 1/100 LC50, chlorpyrifos, imidacloprid and the fungal product caused no deaths.
Chlorpyrifos was the only toxicant that significantly decreased the activity of brain AChE. Concerning the effect on brain total esterase, chlorpyrifos, imidacloprid and neem oil significantly decreased the activities of this enzyme while cypermethrin, mineral oil and the fungal product were of no effect.
For the effect on liver function, chlorpyrifos, cypermethrin and neem oil were found to significantly alternate the activities of alkaline phosphatase, transaminases and other biochemical measurements. All the non-conventional insecticides showed no effects on bilirubin concentrations. It is generally noticed that results of the effects on biochemical markers in fish are in parallel with those obtained on quail birds with the exception of imidacloprid that showed no significant biochemical lesions in fish and this could be explained on the basis of photodegradation of imidacloprid in water. It thus obvious that biochemical markers of liver function of Bolti fish have not been adversely affected by the tested non-conventional insecticides unlike chlorpyrifos, cypermethrin and neem oil that showed liver dysfunction. The harmful effects were less pronounced at the concentration, 1/100 LC50. However, dysfunction might be reversible and curable after non-exposure period to the toxicants.
All the tested non-conventional insecticides showed no significant alterations in levels of creatinine and urea. However, imidacloprid and the mineral oil were found to elevate creatinine at 1/50 LC50 chlorpyrifos and cypermethrin were the most dangerous ones since they affected urea and creatinine levels at the two tested concentrations. Cypermethrin is less powerful than chlorpyrifos being of no effect on urea level at 1/100 lC50. It could be concluded that chlorpyrifos might be nephrotoxic and the rest of materials especially neem oil and the fungal product might be of low renal toxicity.
Concerning the effect on sugar content, none of the conventional insecticides was of significant effect on glucose level in viscera of fish homogenates.
In general, the study suggests that the biorational insecticides especially the mineral oil and the entomopathogenic fungal product might be toxicologically safer than the conventional synthetic ones. Comprehensive toxicological and ecotoxicological studies on different animal and avian species are further needed to verify the overall impact of these biorationals and possibly their chemical constituents on ecosystem using more realistic exposure conditions especially for fish and other marine organisms.
This book provides a concise synthesis of how toxic chemical pollutants affect physiological processes in teleost fish. This Second Edition of the well-received Water Pollution and Fish Physiology has been completely updated, and chapters have been added on immunology and acid toxicity. The emphasis, as in the first edition, is on understanding mechanisms of sublethal effects on fish and their responses to these environmental stressors. The first chapter covers the basic principles involved in understanding how fish respond, in general, to environmental alterations. Each subsequent chapter is devoted to a particular organ system or physiological function and begins with a short overview of normal physiology of that system/function. This is followed by a review of how various toxic chemicals may alter normal conditions in fish. Chapters covering environmental hypoxia, behavior, cellular enzymes, and acid toxicity are also included. The book closes with a discussion on the practical application of physiological and biochemical measurements of fish in water pollution control in research and regulatory settings.
In this work, shearing interface coassembly in biliquid phase systems is employed to synthesize biocompatible core-shell magnetic mesoporous silica microspheres with uniform size of about 600 nm, perpendicular mesopores of 6.0 nm and large pore volume of 0.77 cm³/g. The toxicology assays based on the zebrafish model was conducted to test under a high-throughput manner for the biosafety of Fe3O4@[email protected]2 microspheres. The highest no observed toxic effect concentration (NOEC) estimated by the acute toxicity assay for the microspheres was 1.6 mg/mL. The estimated number (measured by ICP-MS) of the penetrated microspheres at this concentration was 2 × 10⁶ per embryo. The results of three different performed toxicity assays show no overall acute toxicity, teratogenicity, or neurotoxicity of the microspheres on zebrafish embryos at any of the tested concentrations (from 0.1 to 1.6 mg/mL) via its multifunctional microstructure. Here, gemcitabine (GEM) as a model of anti-cancer drug was loaded into the mesopores of Fe3O4@[email protected]2 microspheres to study their drug release behavior. The microspheres were found to exhibit pH responsive property, which benefits for the GEM release under cancer therapy. Overall, this study offers promising avenue for effective evaluation of magnetic core-shell microspheres for drug delivery and cancer therapy applications.
Die Reaktion eines Organismus auf eine toxische Verbindung muß als eine allgemeine Reaktion-Stimulus-Kurve beschrieben werden, wie sie in nahezu allen biologischen Systemen auftritt. Es ergibt sich eine rechtsschiefe Verteilungsfunktion, die durch eine einfache logarithmische Transformation des Stimulus (= Konzentration oder Kontaminationsdauer des Toxins) in eine Normalverteilung überführt werden kann. Diese Normalverteilung wird durch zwei Parameter definiert, den Mittelwert der Normalverteilung (d.h. 50% der Organismen zeigen eine Reaktion) und die Standardabweichung der Normalverteilung als Varianz der Reaktion (der Mittelwert ± Standardabweichung umfaßt den Bereich 16 bis 84%). Als statistisches Verfahren zur Beschreibung von toxischen Wirkungen auf Organismen dient die Wahrscheinlichkeitsanalyse (= Probitanalyse, Finney 1971), und es können statistisch gesicherte Daten über die Toxizität von Stoffen ermittelt werden. Als Parameter werden erhoben:
LC50 („median lethal concentration“) als mittlere letale Konzentration; der Index 50 gibt den Prozentsatz der geschädigten Organismen an. Mitunter werden auch andere Wirkungsschwellen betrachtet, z.B. LC10 für 10% geschädigte Tiere. Allerdings nimmt die Signifikanz der Daten in den Randbereichen der Normalverteilung deutlich ab. Bei der Erfassung der LC50 ist die Expositionszeit der Standardtestverfahren mit 1 bis 2 Tagen vorgegeben.
EC50 („median effective concentration“) als mittlere subletal wirksame Konzentration; in einigen Testverfahren ist es notwendig, subletale Effekte zu erfassen, da der Zeitpunkt des Todes nur sehr schwer ermittelbar ist (u.a. bei Wasserflöhen Daphnia). Bei der Angabe einer EC50 muß der erfaßte Effekt, z.B. Immobilisierung der Daphnien, mit benannt werden.
LT50 („median lethal time“) beschreibt die mittlere Zeit bis zum Eintritt einer letalen Wirkung; bei der Angabe einer LT50 muß die entsprechende Konzentration des Toxins vorgegeben werden, z.B. aufgrund der Vorflutverhältnisse. Die LT50 liefert somit eine toxikologische Bewertung einer speziellen Gewässersituation.
ET50 („median effective time“) als mittlere Zeit bis zum Eintritt eines subletalen Effektes. Bei Angabe einer ET50 muß die entsprechende Konzentration des Toxins und der erfaßte Effekt benannt werden.
Malathion is an organophosphate insecticide which uses to destroy insects and pests of fruit trees, ornamental plants and agricultural corps.In the present study, effect of Malathion on liver and selected enzymes (SGOT, SGPT and ALP) was studied in Caspian Roach (Rutilusrutiluscaspicus). Four treatments with three replications were designed to carry out the survey. Four groups of experimental fish (containing 30 fish in each group) were exposed to different concentrations of Malathion. e. 0, 0.01, 0.05 and 0.1 ppm respectively for 23 days. Blood collection was done in 3rd, 13th and 23th after exposure to Malathion and also 30 days after recovery in clean water and enzymes were measured using standard kits. Also liver tissues were isolated to histological examination. Results showed that tissues of control group (0ppm) were normal and there were no damages, yet there were hepatocytes degeneration, picnotic in nuclear, hepatocytes vacuolization, vascular congestion and sinusoid congestion in liver of other groups. Tissue damages were increased in higher malathion concentration and over time. Results related to enzymes showed that there were no significant differences in SGOT of fish treated with low concentrations of malathion (0.01 and 0.05 ppm) and control group but it was increased in highest concentration (P<0.05). Yet, SGPT increased significantly after passing 23 days in all fish exposed to malathion But ALP changes trend was decreasing.
Histopathologic lesions in the liver of juvenile Prochilodus lineatus were assessed following 24 and 48h exposure to the organophosphate pesticide Dipterex 500 ® (Trichlorfon) at a concentration of 0.2 µl/L. Livers of experimental animals were collected, processed, and stained with hematoxylin and eosin according to routine histology methods. Alterations in hepatic tissue were observed after pesticide exposure, including lateral migration of nuclei, variation in the diameter and density of nuclei, pyknosis and necrosis.
To assess the effects of hexavalent chromium on acid and alkaline phosphatase in six vital tissues (gills, kidney, intestine, liver, brain and muscle) fishes (P. dipes) were exposed to three sublethal concentrations of Cr (VI) (5, 10 and 15 mg/l) for three different exposure durations (2,4 and 6 days). The activity of both the enzymes showed dose and exposure duration dependent inhibition in almost all the tissues studied. However, in kidney, liver and intestine the enzyme activity was stimulated. It was evident that both dose and duration dependent alteration of enzyme activity occurred in gills and kidney. However, in other tissues predominantly exposure duration dependent alterations were observed.
The effects of sodium fluoride were investigated on the biochemical parameters in Heteropneustes fossilis. This study was carried out to evaluate the toxicity of fluoride at three levels (25mg/L,50mg/L and 75mg/L) for 90 days. The biochemical changes in plasma showed significant increase in glucose, total protein, triglyceride, cholesterol, SGOT, SGPT, and ALP. These results indicate that sodium fluoride can alter the plasma's biochemical contents of H. fossilis. Thus it can be concluded that fishes can effectively used as monitors of water quality with respect to fluoride as well as toxicants.
Neste trabalho foram avaliadas as alterações histopatológicas de fígados de juvenis de curimbatá, Prochilodus lineatus (Valenciennes, 1836) expostos a uma concentração de 0,2 µl/L do organofosforado Dipterex 500 (Trichlorfon). Após 24 e 48 horas de exposição coletaram-se os fígados dos peixes contaminados e controle, que foram processados pelas técnicas de rotina para histologia e corados pela hematoxilina e eosina (HE). As alterações hepáticas observadas foram: migração lateral do núcleo dos hepatócitos, núcleos com vários diâmetros e diferentes graus de densidade, picnose e necroseHistopathologic lesions in the liver of juvenile Prochilodus lineatus were assessed following 24 and 48h exposure to the organophosphate pesticide Dipterex 500 ® (Trichlorfon) at a concentration of 0.2 µl/L. Livers of experimental animals were collected, processed, and stained with hematoxylin and eosin according to routine histology methods. Alterations in hepatic tissue were observed after pesticide exposure, including lateral migration of nuclei, variation in the diameter and density of nuclei, pyknosis and necrosis
The input of agrochemicals in the aquatic compartment can results in biochemical injuries for living organisms. In this context, the knowledge of alterations of enzymatic activities due the presence of agriculture pollutants contributes for the elucidation of the mechanisms of toxicity, implementation of economic methods for monitoring purposes and establishment of maximum allowed concentrations. In the present work, the above considerations are discussed, and data concerning changes in enzymatic function by pesticides and fertilizer contaminants are reviewed. Also, we focused on the acid phosphatase due its susceptibility to several pollutants and diversity in cellular functions.
Glyphosate is a broad-spectrum organophosphate (OP) herbicide, highly soluble in water, and when applied in terrestrial systems it penetrates into soil, eventually reaching the aquatic community and affecting nontarget organisms. The aim of this study was to evaluate the toxicity of glyphosate on ovaries of zebrafish (Danio rerio). Ovaries (n = 18 per triplicate) were exposed to 65 μg/L of glyphosate [N-(phosphonomethyl) glycine] for 15 d. This concentration was determined according to Resolution 357/2005/CONAMA/Brazil, which establishes the permissible concentration of glyphosate in Brazilian inland waters. Nonexposed ovaries (n = 18 per triplicate) were used as control. Subsequently, morphology and expression of steroidogenic factor-1 (SF-1) of exposed and nonexposed ovaries was determined. No apparent changes were noted in general morphology of exposed and nonexposed ovaries. However, a significant increase in diameter of oocytes was observed after exposure to glyphosate. When ovarian ultrastructure was examined the presence of concentric membranes, appearing as myelin-like structures, associated with the external membranes of mitochondria and with yolk granules was found. After glyphosate exposure, immunohistochemistry and immunoblotting revealed greater expression of SF-1 in the oocytes, which suggests a relationship between oocyte growth and SF-1 expression. These subtle adverse effects of glyphosate on oocytes raised a potential concern for fish reproduction. These results contribute to understanding glyphosate-induced toxicity to nontarget organisms, showing subcellular and molecular impairments that may affect reproduction in +female fish.
Among the new substances that were synthesised by the chemical industiy during the last decades, with the aim to improve human prosperity, some cause negative effects on non target organisms like fish. Among the pesticides, the organophosphates (OP) are widely used in cattle management, agriculture and aquaculture activities. Brachydanio rerio Hamilton-Buchman, 1822 is used world-wide in fish-tests for the establishment of the lethal levels of pesticides. Nevertheless, these fish show morphological changes in their hepatocytes when exposed experimentally for 24 hours to the sublethal dose of 0.025 µl/l of the OP Dimethoate 500, in acute tests. Right hours after exposure, a high density of cells with cytoplasmic granulation is seen as well as focal necrosis. The main changes that occurred in the first 24 hours of exposure were the loss of the typical polygonal cell shape and of detectable cell limits, lateral migration of nuclei, nuclear size and shape, condensation of chromatin and pycnosis, increased cytoplasmic granulation followed by vacuolisation. After 48 hours in clean water, 72 hours from contamination, besides significant morphological changes and necrosis, focal regeneration of the hepatocytes, canaliculi, ducts and sinusoids were seen. Even having been exposed to the half of the dose considered as safe according to the general pesticide allowance rules, significant changes of the liver tissue were observed.
The input of agrochemicals in the aquatic compartment can results in biochemical injuries for living organisms. In this context, the knowledge of alterations of enzymatic activities due the presence of agriculture pollutants contributes for the elucidation of the mechanisms of toxicity, implementation of economic methods for monitoring purposes and establishment of maximum allowed concentrations. In the present work, the above considerations are discussed, and data concerning changes in enzymatic function by pesticides and fertilizer contaminants are reviewed. Also, we focused on the acid phosphatase due its susceptibility to several pollutants and diversity in cellular functions.
This study was conducted to investigate the induction of biochemical stress responses in Macrobrachium malcolmsonii following exposure to endosulfan and to determine the most sensitive parameter of endosulfan-induced stress in this species of prawn. Intermolt juvenile prawns were exposed to three sublethal concentrations of endosulfan (10.6, 16.0, and 32.0 ng/L) for a period of 21 days. Samples were taken from the hemolymph, brain, hepatopancreas, gills, and muscle of respective prawns from each test and control group on 1, 8, 15, and 21 days of exposure. The content of glutathione S-transferase (GST) was found to be higher in test prawns than in controls. This suggests that there was an activation of a mechanism to detoxify endosulfan. However, the toxic effects of endosulfan were not fully neutralized, and, hence, there was evidence of impairments in various biochemical mechanisms. Biphasic alterations were noted in phosphatases and in lactate dehydrogenase (LDH) levels in tissues of test prawns. The concentration of soluble protein in the hemolymph was found to be elevated, whereas this declined in other tissues of test prawns in comparison to controls. Elevation in the level of total free amino acid and decline in DNA and RNA levels in test prawns were recorded. Endosulfan toxicity resulted in breakdown of various polypeptides in tissues of test prawns. Thus, protein denaturation appeared to be one of the manifested toxic effects of endosulfan. The content of acetylcholinesterase (AchE) declined in test prawns. This reflects the disturbances of basic metabolic activities in test prawns. The grade of endosulfan-induced biochemical stress responses in test prawns was in the order phosphatases > LDH > GST > protein denaturation > AchE. The present investigation records that endosulfan severely impairs various biochemical and physiological mechanisms in M. malcolmsonii and hence poses a threat to its survival, growth, and maturation.
Experiment,s were carried out on the toxicity of diluted dyeing and printing industry
effluent to a penaeid prawn, Parapeimeopsis sculptilis (HELLER). Effluent tolerance experiments
showed that bot,h juvenile and adult prawns were unable to withstand the increasing concentration
of the effluent. Juveniles were more susceptible to effluent than adults. As a consequence
of higher consumption of energy due to the disturbed metabolism a decrease in the metabolite
content was observed, whereas the content of ions showed an irregular trend with all the concentrations.
The behavioural st'udy showed that before death both juvenile and adult prawns became
inactive, followed by paralysis. The physico-chemical nature of the effluent showed high amounts
of solicls? BOD. COD, sodium, magnesium, calcium, chloride. ammonium nit.rogen. sulphate and
absence of DO, probably causing an osmotic imbalance in the prawn body which leads to early
mortality. Even a 1 concentration of such an effluent of a physico-chemical nature was reported
to bc toxic to the pra'wns
Typescript (photocopy). Thesis (M.S.)--Oregon State University, 1991. Includes bibliographical references (leaves 122-133).
Four-month old adult siblings of zebrafish were exposed to four concentrations of diazinon for up to 168 h. DNA, RNA, protein and total free amino acid content were monitored in the liver. The DNA, RNA and protein contents were significantly reduced, whereas the amino acid content was significantly enhanced. All these changes showed dose- as well as time-dependent response.
Signal transduction by xenobiotics in fish has recently gained much attention. The better known transduction mechanisms are those elicited by organochlorines, organophosphates, carbamates and heavy metals. Organochlorines specifically bind to the membrane bound ouabain sensitive Na+-K+-ATPase affecting neural transmission while the organophosphates and carbamates bind specifically to the membrane bound enzyme acetylcholinesterase again affecting neural transmission. Since the nervous system is one of the important integrative and interactive physiological systems in animals, hypofunction of the nervous system leads to secondary effects in the endocrine system including thyroidal, gonadal, interrenal, pituitary and hypothalamic functions. Even low levels of xenobiotics are efficient enough to bring about remarkable changes in the functional physiology of the non target animals. Heavy metals such as cadmium or mercury belonging to the same group II B in the periodic table probably have a similar mechanism of action. Avidity of these metals to SH-radicals allow them to bind indiscriminately to SH groups in proteins. One pathway of interaction by inorganic mercury with the membrane bound ouabain sensitive Na+-K+-ATPase has been clearly established in fish liver and ovary. Binding of inorganic mercury to the membrane bound enzyme is through sulfhydryl group which inactivates the sodium pump leading to accumulation of the cation in the cytosol. The inorganic mercury is next conjugated by the cytosolar nucleophile, glutathione, and is transported to the nucleus where dissociation occurs and the free metal binds to the metal regulatory element to initiate gene expression. The inducible proteins are 3beta-hydroxysteroid dehydrogenase in the oocyte and metallothionein and C-reactive protein in the liver. The present review deals with the role of xenobiotic as a stress factor.
The zebrafish (Danio rerio) is now the pre-eminent vertebrate model system for clarification of the roles of specific genes and signaling pathways in development. The zebrafish genome will be completely sequenced within the next 1-2 years. Together with the substantial historical database regarding basic developmental biology, toxicology, and gene transfer, the rich foundation of molecular genetic and genomic data makes zebrafish a powerful model system for clarifying mechanisms in toxicity. In contrast to the highly advanced knowledge base on molecular developmental genetics in zebrafish, our database regarding infectious and noninfectious diseases and pathologic lesions in zebrafish lags far behind the information available on most other domestic mammalian and avian species, particularly rodents. Currently, minimal data are available regarding spontaneous neoplasm rates or spontaneous aging lesions in any of the commonly used wild-type or mutant lines of zebrafish. Therefore, to fully utilize the potential of zebrafish as an animal model for understanding human development, disease, and toxicology we must greatly advance our knowledge on zebrafish diseases and pathology.
The present study was performed to determine the toxic effects of endosulfan on the quantitative and qualitative aspects of acid and alkaline phosphatases (ACP and ALP) in Macrobrachium malcolmsonii. Intermoult juvenile prawns were exposed to 32.0 ng/l of endosulfan for a period of 21 days. Samples were taken from the hemolymph, hepatopancreas, gills and muscle of test prawns on the 21st day. The content of ACP and ALP in the hepatopancreas of test prawns were found to be higher in comparison to respective controls. The levels of these enzymes in the hemolymph, gills and muscle of test prawns were found to be lower than in the same tissues of controls. In non-denaturing PAGE, phosphatases appeared as white bands. The intensity of white bands in the hepatopancreas of test prawns were found to be higher when compared to controls. In the hemolymph, gills and muscle of test prawns, the intensity of white bands were found to be lower in comparison to controls. The results of the present study suggest that endosulfan affects the quantity and quality of ACP and ALP in the tissues of M. malcolmsonii.
The protective role of sesamol, an inhibitor of the mixed function oxygenase (MFO) system, against histopathological effects of methyl parathion in the hepatopancreas of the estuarine crab, Chasmagnathus granulatus, was studied. Exposure (72 h) to a sublethal dose (0.05 mg/kg/day; 10% of 72 h-LD50) of injected methyl parathion increased the percentage of damaged hepatopancreatic tubules. Presence of melanin-like deposits in the connective tissue between hepatopancreatic tubules was also observed. Antioxidant enzyme activities (catalase and glutathione S-transferase) and lipid peroxidation (LPO) levels were also increased in hepatopancreas of crabs injected with methyl parathion. Pretreatment with sesamol (0.85 mg/kg/day) significantly protected against all these effects. These findings suggest that the hepatopancreatic damages induced by methyl parathion are due to LPO of hepatopancreatocytes membranes, as a consequence of the oxidative stress generated after methyl parathion oxidative biotransformation mediated by the MFO system.
The procedure for the determination of nucleic acids described in this chapter is based on the finding that nucleic acids can be separated from other tissue compounds by their preferential solubility in hot trichloroacetie acid. The isolated nucleic acids are then quantitated by means of colorimetric reactions involving the pentose components of the nucleic acids. The determination of nucleic acids in tissues is largely a problem in identification. By means of the extraction procedures described in the chapter and the colorimetric reactions of peptide nucleic acid and DNA, a considerable degree of specificity is placed on the determination of these compounds. Occasionally however, false results will be obtained, owing to the presence of materials in the nucleic acid extracts that interfere with the pentose reactions. It is emphasized that the extraction methods described were developed for nucleic acid determinations by spectrophotometric methods. Although it was at first thought that these procedures might be directly applicable to isotopic work, it has become quite clear that the separations are not sufficiently refined for such studies. The methods have served, however, as starting points for other separation procedures more suitable for isotopic work.
Toxicity tests reveal that the zebrafish (Brachydanio rerio) is very sensitive to malathion, an organophosphorus insecticide, and 24 to 96-h LC50 values showed a gradual decrease as the exposure time was increased. Zebrafish exposed to long-term sublethal concentrations failed to spawn and invariably exhibited skeletal deformities. Possible causes of the reduced gonadosomatic index and skeletal deformities are discussed.
The synergistic effects of piperonyl butoxide (PB)2, sulfoxide (SU), and dimethyl amino aniline (DAA) used in conjunction with two organophosphate pesticides (phorate and formothion) and two carbamate pesticides (mexacarbate and carbaryl) were examined in terms of the LC50 and acetylcholinesterase (AChE) inhibition in the snailLymnaea acuminata. The three synergists reduced the LC50 of phorate, formothion, mexacarbate, and carbaryl at exposure periods ranging from 24 to 144 hr. PB and SU had strong synergistic action; DAA had little effect; the highest synergistic ratio (70) was found with carbaryl and PB. The synergists enhanced thein vitro inhibition of cholinesterase caused by the pesticides; the synergistic effect onin vitro inhibition, however, was lower, compared to the change in the LC50.
The snailLymnaea acuminata, vector of the liver flukesFasciola hepatica andFasciola gigantica, was exposed to 40% and 80% LC50 doses of the pesticides 12/24 mg/L phorate (O,O-diethylS-(ethylthio) methyl phosphorodithioate) and 2.5/5.0 mg/L of mexacarbate (4-dimethylamino)-3,5-dimethylphenyl methyl carbamate) for 24 or 48 hr. These pesticides, although considered to be antiacetylcholinesterase agents, inhibited alkaline phosphatase and enhanced the activity of acid phosphatase in hepatopancreas, mantle, intestine, and foot of the snail. Trypsin activity in the alimentary canal of the snail was also inhibited. Discontinuation of pesticide treatment resulted in partial recovery in mexacarbate-treated animals while in phorate-treated animals the effect of the pesticide was largely irreversible.
Oral administration of a dose of leptophos which does not produce delayed neurotoxicity to hens caused significant inhibition of plasma cholinesterase. The enzymatic activity which was lowest one half day after the administration gradually increased with time and was completely recovered on day 20. Plasma acid phosphatase activity significantly increased following the oral administration of leptophos, and the enzymatic activity was significantly higher than that of the control throughout the experiment. The increase in acid phosphatase activity is implicated in the mechanism of delayed neurotoxicity caused by leptophos in hens.
The effect of LD50 (1.8 mg/liter) and a sublethal concentration (0.3 mg/liter) of mercuric chloride on the digestive system of a teleost fish, Channa punctatus, has been studied at intervals of 96 hr and 5, 10, 15, 20, and 30 days. The results show that drastic changes are produced in the histological structure of the liver. These include liver cord disarray, connective tissue damage, granulation and vacuolation of the cytoplasm and hypertrophy of the nucleus, necrosis, fatty infiltration, proliferation of connective tissue, glycogen depletion, and cirrhosis. In the digestive system, treatment produced more drastic changes, especially between 15 and 30 days of treatment. The gastric gland cells showed a decrease in pepsinogen granules, followed by degeneration of the entire cells. Pyloric caeca were the most affected portion of the alimentary canal. The villi in this organ and in the intestine were ruptured. Lateral cell boundaries had disintegrated, and cells presented a syncytial appearance. Inflammatory cells collected at a number of places in the mucosa. In the intestine, initially, a hyperactivity of mucus secreting goblet cells was observed. This was followed by release of large amount of mucus into the intestinal lumen. No conspicuous change was observed in the rectal portion.
Acetylcholinesterase (AChE) activity in the nervous tissue (brain) of the fish Brachydanio rerio was significantly inhibited by exposure to the organophosphorus (OP) pesticide, malathion. The inhibition was dose- as well as time-dependent. The fish survived even when the activity of the enzyme was inhibited by 90%. There was a significant recovery in the activity of AChE when malathion stress was lifted.
The effect of various concentrations of thiodon on tissue non-specific phosphomonoesterases was studied in 9 edible freshwater teleosts from a tropical environment. Sublethal doses (5 and 10 ppm) of thiodon increased alkaline phosphatase in liver, muscle, kidney and brain, while lethal doses (15 and 20 ppm) decreased it. The reverse was the case with acid phosphatase. The greatest changes were recorded in kidney and liver. The increase in the activity of acid phosphatase after lethal doses might be attributable to rupture of the cellular and lysosomal membranes. The decrease in alkaline phosphatase activity might be related to the fall in pH following rupture of these membranes. The changes were greater in major carp species (C. catla, L. rohita and C. mrigala) than in catfish (H. fossilis, C. batrachus and M. seenghala) or snakehead species of murrels (C. striatus, C. punctarus and C. marulius).
Thirty years of testing has yielded over 120 esterase inhibitors in current use for pest control. Several hundred million pounds of these organophosphates and carbamates are employed each year as insecticides and acaricides and, to a much lesser extent, as anthelmintic agents, nematocides, and herbicides. Systemics or chemotherapeutic agents for control of insect pests of plants and animals first became practical with the organophosphates. Compounds of lower mammalian toxicity and other favorable biological properties continue to appear and displace established compounds and broaden the use areas. Problems of resistance and residues in certain areas of insect control by chlorinated hydrocarbons will result in a further shift to esterase-inhibitors for pest control. Interpretation of the potential hazards of pesticides to man is dependent on the availability of fundamental information on their modes of action combined with use experience; this knowledge is available for the organophosphates and carbamates that act as acetylcholinesterase inhibitors.
Since 1922 when Wu proposed the use of the Folin phenol reagent for
the measurement of proteins (l), a number of modified analytical pro-
cedures ut.ilizing this reagent have been reported for the determination
of proteins in serum (2-G), in antigen-antibody precipitates (7-9), and
in insulin (10).
Although the reagent would seem to be recommended by its great sen-
sitivity and the simplicity of procedure possible with its use, it has not
found great favor for general biochemical purposes.
In the belief that this reagent, nevertheless, has considerable merit for
certain application, but that its peculiarities and limitations need to be
understood for its fullest exploitation, it has been studied with regard t.o
effects of variations in pH, time of reaction, and concentration of react-
ants, permissible levels of reagents commonly used in handling proteins,
and interfering subst.ances. Procedures are described for measuring pro-
tein in solution or after precipitation wit,h acids or other agents, and for
the determination of as little as 0.2 y of protein.
Metabolism of leptophos by liver microsomal enzymes: A factor in species selectivity to delayed neurotoxicity Verlag Chemie Record of Zebra danio (Brachydanio rerio) (Cyprinidae) from Uttar Pradesh with notes on sexual dimorphism
- M B And Ashry
- M A And Szcypinski
- A J Kumar
ABOU-DOMA, M. B., AND ASHRY, M. A. (1978). Metabolism of leptophos by liver microsomal enzymes: A factor in species selectivity to delayed neurotoxicity. Fed. Proc. 37,504. ANDERSCH, M. A., AND SZCYPINSKI, A. J. (I 947). Amer. J. Clin. Pathol. 17, 57 1. Cited in Methods of Enzymatic Analysis (H. V. Bergmeyer, ed.) (1967). Verlag Chemie, Deer6eld Beach, Fla. ANSARI, B. A., AND KUMAR, K. (1982). Record of Zebra danio (Brachydanio rerio) (Cyprinidae) from Uttar Pradesh with notes on sexual dimorphism. J. Adv. Zool. 3,88-89.
Text Book ofPhysiology and Biochemistry, pp. 103-105. TinlingandCo
- G H Bell
- J N Davidson
- D E And Smith
BELL, G. H., DAVIDSON, J. N., AND SMITH, D. E. (1970). Text Book ofPhysiology and Biochemistry, pp. 103-105. TinlingandCo.,London. BERGMEYER, H. V. (ed.) (1974). Methods ofEnzymatic Analysis (modi6ed from Andersch, M. A., and Szcypinski, A. J. (1947). Amer. J. Clin. Pathol. 17,57 1). Verlag Chemie, Deerfield Beach, Fla. C~~1~~,J.E.(1964).Esteraseinhibitorsaspesticid~.Science146, 1011-1017.
Toxic effects of endrin on liver and kidney of a teleost fish Determination of nucleic acids in tissues by pentose analysis
- K V Sastry
- S K And Sharma
SASTRY, K. V., AND SHARMA, S. K. (1979). Toxic effects of endrin on liver and kidney of a teleost fish. In Proceedings Symp. Environ. Biol., pp. 337-342. Muzathunagar, India. SCHNEIDER, W. C. ( 1957). Determination of nucleic acids in tissues by pentose analysis. In Methods in Enzymology (S. P. Colowick and N.
Methods of Enzymatic Analysis
- Andersch
Labilization of lysosomes as an aspect of the biochemical toxicology of anticholinesterase pesticides
- Ntiforo
- Andersch
Toxic effects of endrin on liver and kidney of a teleost fish
- Sastry
Effect of phosphamidon in a fresh water teleost fish Nemachelius denisonii (Day). Histopathological and biochemical studies
- Rashatwar
Metabolism of leptophos by liver microsomal enzymes: A factor in species selectivity to delayed neurotoxicity
- Abou-Donia