Comparative assessment of acute toxicity and histological changes in liver of African catfish Clarias gariepinus exposed to cotton insecticides.

This study investigated the acute toxicity in Clarias gariepinus to insecticides currently used in Benin cotton fields, including Thalis 112 EC (Emamectin benzoate 48 g L-1, Acetamiprid 64 g L-1), Vizir C 92 EC (Cypermethrin 72 g L-1, Abamectin 20 g L-1), Pyrinex Quick 212 EC (Deltamethrin 12 g L-1, Chlorpyrifos 200 g L-1) and Pyro FTE 472 EC (Cypermethrin 72 g L-1, Chlorpyrifos 400 g L-1) with emphasis on liver histopathological effects. A set of 180 juveniles of C. gariepinus (5.21 ± 3.22 g) was exposed for 96 h to increasing concentrations of each pesticide. The values of 96-hLC50 were 4.778, 0.002, 0.004, and 0.012 µL L-1 for Thalis, Vizir, Pyrinex, and Pyro, respectively, indicating that Vizir, Pyrinex, and Pyro were very highly toxic to C. gariepinus juveniles. During the experiments, the morphological and behavioral responses (discoloration, hyperactivity, lethargy, etc.) were observed in exposed fish, hypothesizing the neurotoxicity of these pesticides. Histopathological alterations observed in liver of contaminated fish were regressive changes, such as necrosis, vacuolation, bleeding, nuclear degeneration, hepatocytes degeneration, sinusoids dilatation, etc. Vizir induced the highest histological alteration indices while the lowest were induced by Thalis, confirming the highest toxicity of Vizir. These results indicate that acute concentrations of these insecticidal molecules have destructive effects on the liver of C. gariepinus.

Evaluation of acute toxicity and histology effect on liver of glyphosate and atrazine in the African catfish Clarias gariepinus (Burchell 1822).

Aquatic organisms are exposed to chemical pesticides including glyphosate (Sharp 480 SL) and atrazine (Atraforce), two phytocidal molecules used for agriculture purposes in Benin. In this study, we assessed the acute toxicity of these two herbicides with emphasis on their histopathological effects on the liver of catfish Clarias gariepinus. One hundred and eighty juveniles of C. gariepinus (mean length 7.26 ± 0.59 cm and mean weight 5.21 ± 3.22 g) were exposed over 96 h to increasing concentrations of each phytocide. The values of 96 h-LC50 were 6.175 × 103 and 3.165 ppm, respectively for Sharp 480 SL and Atraforce. This indicates that Sharp 480 SL was nontoxic, while Atraforce displayed a moderate toxicity to C. gariepinus juveniles. During the tests, the behavioral responses (hyperexcitation, lethargy, loss of balance, discoloration of skin, etc.) that usually precede death were observed in exposed fishes, confirming the neurotoxicity of these phytocides. Histological alterations observed in liver of contaminated fishes were regressive changes, such as necrosis, hepatocyte vacuolation, nuclear degeneration, hepatocytes degeneration, sinusoids dilatation, etc. These results indicate that exposure to these herbicides had destructive effects on the liver of C. gariepinus.

Chronic effects of a binary insecticide Acer 35 EC on Nile tilapia Oreochromis niloticus through a multi-biomarker approach.

Acer 35 EC is a widely used insecticide (a binary mixture of lambda-cyhalothrin and acetamiprid) in pest control in many West African countries, particularly in the cotton culture in north Benin. The aim of this study was to investigate the chronic effects of Acer 35 EC on Nile tilapia Oreochromis niloticus juveniles using a multi-biomarker approach under laboratory conditions. For this purpose, fish were exposed to sublethal concentrations of Acer 35 EC (0, 1 and 10% of LC50- 96 h value). After 28 and 56 days of exposure, several biomarkers were measured in males and females including enzymatic activities related to detoxification and oxidative stress, neurotoxicity and immune responses, sex steroid hormones (testosterone, 17ß-estradiol and 11-keto-testosterone) and histological alterations of liver, kidney and gonads. An Integrated Biomarker Response (IBR) was then calculated. The results showed a reduction of cholinesterase activity in muscles, and intercellular superoxide anion production in both sexes. Female steroidogenesis and gametogenesis were affected, especially testosterone levels and oocyte growth. More alterations were observed in liver after exposure to Acer 35 EC. In both sexes, IBR values were higher after 56 days than after 28 days of exposure. In conclusion, based on a large set of biomarkers and IBR values, the chronic exposure to low doses of insecticide Acer 35 EC seems to impair different physiological functions in Nile tilapia juveniles on a time-dependent manner, with a stronger impact on females than on males.

Joint toxicity of two phytosanitary molecules, lambda-cyhalothrin and acetamiprid, on African catfish (Clarias gariepinus) juveniles.

This study was designed to evaluate the acute toxicity of acetamiprid (neonicotinoid) and lambda-cyhalothrin (pyrethroid), individually and mixtures (Act-LCh mixture and Acer 35 EC®, 15 g/L of acetamiprid and 20 g/L of lambda-cyhalothrin) in African catfish juveniles (3.35 ± 0.75 g). The tests were conducted in the laboratory under semi-static conditions according to OECD Guideline 203. Mixture toxicity effects as a function of lethal concentrations were assessed using the additive index (AI) method. Acetamiprid with 96 h-LC50 = 265.7 ppm can be considered to be nontoxic for this species. However, lambda-cyhalothrin was highly toxic to C. gariepinus with 96 h-LC50 = 0.00083 ppm. Acer 35 EC® was less toxic (96 h-LC50 = 0.21 ppm) than the Act-LCh mixture (96 h-LC50 = 0.043 ppm). Marked changes indicating nervous system damage were also recorded. An antagonistic effect was shown for lethal concentrations leading to 5 to 15% mortality in 96 hours (96 h-LC5-15) while an additive effect was obtained for the 96 h-LC20-50. The results indicate that ecological risk assessment of these molecules in aquatic environments should consider their contamination levels. Moreover, particular attention to behavior changes related to their neurotoxicity is recommended for additional monitoring of the negative effects of these insecticides.

Comparative acute toxicity of two phytosanitary molecules, lambda-cyhalothrin and acetamiprid, on Nile Tilapia (Oreochromis Niloticus) juveniles.

This study aimed to compare the toxicity for fish of two active ingredients (lambda-cyhalothrin-20 g L-1, a pyrethroid, and acetamiprid-15 g L-1, a neonicotinoid) which are components of a commercial insecticide (Acer 35 EC) used in cotton crop in many West African countries. The juveniles of Oreochromis niloticus (4.01 ± 0.34 g, mean body weight) were exposed for 96 h to increasing concentrations of active ingredients (lambda-cyhalothrin and acetamiprid) or a mixture similar to Acer 35 EC (composed by 20 g of chemical compound lambda-cyhalothrin and 15 g of acetamiprid dissolved in 1 L of acetone). The experiments were carried out under controlled conditions in aquaria according to OECD Guidelines. During the experiments, the behavioral responses (loss of balance, color change, hyperactivity, etc.) that usually precede death were observed in exposed fish. Mortalities were recorded in each aquarium and the LC50-96h of each chemical was determined. The LC50-96h obtained were respectively 0.1268, 0.0029, 182.9 and 0.5685 ppm for Acer 35 EC, lambda-cyhalothrin, acetamiprid and mixture. All insecticides used in this study had profound impact on Nile tilapia behavior which may confirm the neurotoxicity of each single active compound as well as of their mixture.

Occurrence of Acetamiprid Residues in Water Reservoirs in the Cotton Basin of Northern Benin.

An Ultra performance liquid chromatography (UPLC) coupled to UV detection method was developed to determine acetamiprid residues in water reservoirs of northern Benin, close to cotton fields. The quantification limit of this method was 0.2 µg L-1 acetamiprid in water, its precision ranged between 8% and 22%, and its trueness between 99% and 117% (for concentrations ranging from 0.2 to 5.0 µg L-1). Acetamiprid residues were determined in water samples collected in four reservoirs from northern Benin during the phytosanitary treatment period of cotton. The minimum and maximum concentrations of acetamiprid residues in water were 0.2 and 7.7 µg L-1, respectively. These levels do not represent any risk for human consumption of this water, but indicate a regular use of acetamiprid, possibly together with other pesticides which could be more harmful for both humans and aquatic species.

Genetic characterization of Benin's wild populations of Sarotherodon melanotheron melanotheron Rüppell, 1852.

The Cichlid fish Sarotherodon melanotheron is typically found in West and Central African estuaries and lagoons. It represents a good candidate for promoting tilapia farming in brackish waters. Understanding the genetic diversity in its populations from the hydrographical basins of Southern Benin is primordial before designing selective breeding programs. For this purpose, 202 samples collected from four rivers of Southern Benin and were genotyped using 15 polymorphic microsatellite DNA markers. Each river was split up into three sampling sites. We found significant global linkage disequilibrium across the genome of natural populations of this tilapia species overall the loci. However, when the loci that display aberrant Wright's (FIS and FST) were removed from the data, a linkage disequilibrium was detected for the remaining 11 loci and became compatible with the null hypothesis. Null alleles explained at least 20.58% of FIS variation. We found a significant isolation by distance across subsamples. Effective population size averaged 210 individuals, with a range from 36 to 517 individuals. Assuming that 79% of heterozygote deficits are explained by sib mating lead to a rough estimate of rsm = 0.4 of mating rate between full sibs within S. melanotheron subpopulations. The fish size correlated positively and significantly with the observed FIS (r = 0.58; p value = 0.04806). Reproduction system (endogamy) in S. melanotheron could explain the strong heterozygote deficit observed. Our results provide technical guidance for efficient management of this tilapia species' genetic resources for breeding programs in fresh and brackish waters.

Assessment of the health status of wild fish inhabiting a cotton basin heavily impacted by pesticides in Benin (West Africa).

To determine the impact of agricultural pesticides used in cotton cultivation on the health status of fish living in a Beninese cotton basin, we compared the reproductive and hepatic systems of fish sampled from rivers located in both contaminated and pristine conditions. Different types of biomarkers, including biometric indices (a condition factor K, a gonadosomatic index GSI, and a hepatosomatic index HSI), plasma levels of sex steroids (11-ketotestosterone 11-KT, testosterone T and estradiol-17ß E2) and the histopathology of the gonads and liver, were investigated for two different trophic levels of the following two fish species: the Guinean tilapia Tilapia guineensis and the African catfish Clarias gariepinus. The fish were captured during both the rainy season (when there is heavy use of pesticides on cotton fields) and the dry season from one site, in Pendjari River (reference site), which is located outside the cotton-producing basin, and from three other sites on the Alibori River within the cotton-producing basin. Comparing fish that were sampled from contaminated (high levels of endosulfan, heptachlor and DDT and metabolites) and reference sites, the results clearly indicated that agricultural pesticides significantly decreased K and GSI while they increased HSI, regardless of the season, species and sex of the fish. These pesticides also induced a decrease in the plasma levels of 11-KT and T and increased those of E2. The histopathology of the testes revealed, in both species, a high rate of testicular oocytes, up to 50% in the African catfish, downstream of the Alibori River, which indicated estrogenic effects from the pesticides. The disruption of male spermatogenesis primarily included necrosis, fibrosis and the presence of foam cells in the lobular lumen. The histopathology of the ovaries revealed high levels of pre-ovulatory follicular atresia, impaired oogenesis, a decrease in the oocyte vitellogenic diameter and other lesions, such as fibrosis, vacuolation and melano-macrophagic centers. The histopathology of the liver revealed the presence of necrosis, hypertrophic hepatocytes, foci of vacuolation, glycogen depletion and hemosiderin. An assessment of the general health of the fish indicated that all of the sampled fish from the polluted sites were in poorer health compared with those from the reference site but that the African catfish appeared much more affected than the Guinean tilapia, regardless of the sex and season. In conclusion, the overall results indicated that agricultural pesticides significantly impair the endocrine regulation of fish living in the Beninese cotton basin and that this would most likely be one of the causes of the severe damage observed in the liver and gonads and the reduced health condition.