RESUMO
Pesticides are extensively used for the control of agricultural pests and disease vectors, but they also affect non-target organisms. Cypermethrin (CYP) is a synthetic pyrethroid used worldwide. Otherwise, bioinsecticides like Bacillus thuringiensis var. israelensis (Bti) have received great attention as an environmentally benign and desirable alternative. In order to evaluate the toxicity of those pesticides, Chironomus calligraphus was selected due to its high sensitivity to some toxicants. Third and fourth instars larvae were exposed to serial dilutions of CYP and Bti to determine LC50 values. In order to evaluate the potentially histopathological alterations as biomarkers, after 96-h of exposure, live larvae were fixed for histological analysis of the mid region of digestive tract. The 96-h LC50 values were 0.52 and 1.506µg/L for CYP and Bti, respectively. Midgut histological structure of the control group showed a single layer of cubical cells with microvilli in their apical surface and a big central nucleus. The midgut epithelium of larvae exposed to a low concentration of CYP (0.037µg/L) showed secretion activity and vacuolization while at high concentration (0.3µg/L) cells showed a greater disorganization and a more developed fat body. On the other hand, Bti caused progressive histological damage in this tissue. Chironomus calligraphus is sensitive to Bti and CYP toxicity like other Chironomus species. The histopathological alterations could be a valuable tool to assess toxicity mechanism of different pesticides.
Assuntos
Bacillus thuringiensis/fisiologia , Chironomidae/efeitos dos fármacos , Chironomidae/microbiologia , Larva/efeitos dos fármacos , Larva/microbiologia , Piretrinas/toxicidade , Animais , Núcleo Celular/efeitos dos fármacos , Chironomidae/citologia , Sistema Digestório/citologia , Sistema Digestório/efeitos dos fármacos , Sistema Digestório/microbiologia , Larva/citologia , Microvilosidades/efeitos dos fármacos , Controle Biológico de VetoresRESUMO
The aim of this study was to evaluate the sensitivity of the prawn Palaemon argentinus to the pyrethroid cypermethrin (CYP) and the tetramic acid spirotetramat (STM). These treatments were compared with prawns collected at a reference site to define their basal physiological state. Initially, physicochemical parameters and several pollutants at the selected site were analyzed. The LC50-96 h was determined in adult prawns. Then, prawns were exposed for 96 h to sublethal concentrations of CYP (0.0005 µg/l) and STM (0.44 mg/l) to evaluate the effects on some biochemical endpoints. A treatment combining both pesticides was also added at 5 % of these values. Controls with and without solvent (acetone) were included. The LC50-96 h values were 0.005 µg/l and 4.43 mg/l for CYP and STM, respectively. Moreover, some biomarkers linked to oxidative and energy metabolism were analyzed in the hepatopancreas and muscle of both essayed prawns and those at the basal state. The STM caused a significant decrease in total protein content (32 %) in contrast to the increase of protein carbonyl content (71 %) (p < 0.05). Also, glutathione S-transferase (52 %) and catalase (61 %) activities in the hepatopancreas of exposed prawns were higher compared to both the control and state basal groups (p < 0.05). In muscle, only a significant decrease in the lactate content (69 %) was caused by STM (p < 0.05). In addition, CYP caused a significant increase in the lactate dehydrogenase activity (110 %) in muscle and triacylglycerol content (73 %) in the hepatopancreas (p < 0.05). The integrated biomarker index (IBRv2) analysis showed that STM caused greater damage than CYP. Besides, the combined treatment showed an antagonistic interaction between both insecticides. The differential response of biomarkers to both CYP and STM exposure with respect to their basal levels shows a high sensitivity of P. argentinus demonstrating its potential role as a bioindicator organism.
Assuntos
Biomarcadores , Inseticidas , Palaemonidae , Piretrinas , Compostos de Espiro , Poluentes Químicos da Água , Animais , Palaemonidae/efeitos dos fármacos , Inseticidas/toxicidade , Piretrinas/toxicidade , Compostos de Espiro/toxicidade , Poluentes Químicos da Água/toxicidade , Biomarcadores/metabolismo , Compostos Aza/toxicidade , Hepatopâncreas/efeitos dos fármacos , Hepatopâncreas/metabolismoRESUMO
The aim of this study was to evaluate the toxic effects of the pesticide cypermethrin (CYP) in the digestive gland of the apple snail, Pomacea canaliculata, analysing histological and histochemical alterations. Adult snails were exposed to sublethal CYP concentrations (10, 25, and 100 µg/L) under acute (1 day) and sub-chronic (14 days) conditions. Histological analyses of the morphology of the digestive gland were performed and a histopathological condition index (HI) was calculated. Also, both intracellular accumulation of lipofuscins (LF) and neutral lipids (NL) were evaluated. CYP exposure induced tissue damage to this organ, such as disorganisation of the connective tissue, fibrosis, haemocytic infiltration, atrophy, and necrosis under acute and sub-chronic conditions. These alterations, integrated into a single HI value, revealed notable CYP effects during both acute and sub-chronic exposures. Cell type replacement, measured as VvBAS, was only observed in the sub-chronic treatment. Under acute conditions, the pyrethroid affected NL accumulation at the highest concentration, while in sub-chronic conditions NL accumulation was only observed at the lowest concentrations. P. canaliculata also showed a dose-dependent response of LF under acute CYP exposure conditions. However, under sub-chronic conditions, this parameter was not sensitive to pesticide exposure. All these relevant structural lesions may affect the normal function of the digestive gland, even though the species presented additional mechanisms, as infiltration of hemocyte and basophilic cell hyperplasia, that help it to tolerate the exposure to pollutants. This study showed that some histological and histochemical parameters are sensitive in P. canaliculata at CYP concentrations to which the snail could be exposed in the environments it inhabits.
Assuntos
Poluentes Ambientais , Praguicidas , Piretrinas , Poluentes Químicos da Água , Animais , Poluentes Químicos da Água/toxicidade , Caramujos , Piretrinas/toxicidade , Piretrinas/metabolismo , Praguicidas/metabolismo , Poluentes Ambientais/toxicidade , LipídeosRESUMO
Geraniol (GOH), like other plant-derived natural bioactive compounds, has been found to possess antiproliferative properties that are essential to cope with malignant tumors. However, the mechanisms of molecular action of GOH are not fully elucidated. The aim of this study was to evaluate the effect of GOH on some oxidative parameters in human tumor cell lines (HepG2 and A549). Cytotoxicity evaluated in cell lines by the MTT assay, genotoxicity by the comet assay, and lipid peroxidation by the TBARS. The activities of antioxidant the enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST), were also analyzed. Additionally, intracellular reactive oxygen species (ROS), nitric oxide, and lactate production were determined in HepG2 cells. Both tumor cell lines showed a clear concentration-dependent response to GOH in several of the parameters evaluated. Lipids turned out to be more sensitive than DNA to oxidative damage induced by GOH. TBARS levels increased with respect to control (p < 0.05) by 33% and 122% in HepG2 and A549 cells, respectively treated with 200 µM GOH. However, GOH caused a statistically significant decrease in SOD and CAT activities in HepG2 cells only. GST was not affected in any cell lines. GOH induced the production of ROS but not nitric oxide in HepG2, which shows that ROS were mainly responsible for oxidative damage. Lactate release increased statistically significantly compared to control (p < 0.001), by 41% and 86% at 200 and 800 µM GOH respectively, showing that this monoterpene also affected the glycolytic pathway in HepG2 cells. These results suggest that oxidative stress could mediate the anti-proliferative effects of GOH in HepG2 and A549 cells.
Assuntos
Monoterpenos Acíclicos/farmacologia , Proliferação de Células , Estresse Oxidativo/efeitos dos fármacos , Células A549 , Monoterpenos Acíclicos/administração & dosagem , Monoterpenos Acíclicos/química , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Hep G2 , Humanos , Estrutura MolecularRESUMO
The present study was aimed at evaluating the effect of male presence on ovarian maturation in juvenile females and the role of potential chemical, visual and tactile cues emitted by males in that physiological process. A highly gregarious caridean shrimp with sexual dimorphism, Neocaridina davidi, was used as experimental model. We tested the hypothesis that male presence accelerates ovarian maturation, mainly through chemical cues. Two experiments were performed. In Experiment 1, juvenile females were reared with adult males, adult females or alone, allowing full contact among shrimps. In Experiment 2, these treatments were evaluated allowing chemical and visual communication, only visual communication, or only chemical communication among shrimps. In both experiments juvenile females were observed once a week under a stereomicroscope to determine ovarian growth rate. Although male presence was not necessary for ovarian maturation, it clearly accelerated the rate of ovarian growth, particularly in the last maturation phase. This lead to relatively longer mature ovaries with higher lipid content. On the contrary, the presence of adult females delayed ovarian maturation in juvenile females, while females reared alone showed an intermediate ovarian growth. All these results suggest that adult males release certain cues that stimulate ovarian maturation, while adult females release cues that delay this physiological process. Neither visual cues nor chemical cues released at a distance from females were responsible, either alone or in combination, for the observed effects. Ovarian growth was only influenced when shrimps were allowed to interact freely, probably because of the "mounting" behavior of males towards females. Tactile cues and/or potential chemical cues released by males during this behavior may mediate male stimulatory effect on ovarian growth. Altogether, present results partially support our initial hypothesis and contribute to increase the limited amount of information available on the role of intraspecific multimodal communication in non-behavioral reproductive processes in invertebrate species.
Assuntos
Decápodes/anatomia & histologia , Decápodes/fisiologia , Ovário/crescimento & desenvolvimento , Comportamento Sexual Animal/fisiologia , Maturidade Sexual/fisiologia , Animais , Sinais (Psicologia) , Feminino , Masculino , Ovário/anatomia & histologia , TatoRESUMO
Palaemonetes argentinus, an abundant freshwater prawn species in the northern and central region of Argentina, has been used as a bioindicator of environmental pollutants as it displays a very high sensitivity to pollutants exposure. Despite their extraordinary ecological relevance, a lack of genomic information has hindered a more thorough understanding of the molecular mechanisms potentially involved in detoxification processes of this species. Thus, transcriptomic profiling studies represent a promising approach to overcome the limitations imposed by the lack of extensive genomic resources for P. argentinus, and may improve the understanding of its physiological and molecular response triggered by pollutants. This work represents the first comprehensive transcriptome-based characterization of the non-model species P. argentinus to generate functional genomic annotations and provides valuable resources for future genetic studies. Trinity de novo assembly consisted of 24,738 transcripts with high representation of detoxification (phase I and II), anti-oxidation, osmoregulation pathways and DNA replication and bioenergetics. This crustacean transcriptome provides valuable molecular information about detoxification and biochemical processes that could be applied as biomarkers in further ecotoxicology studies.
Assuntos
Desintoxicação Metabólica Fase II/genética , Desintoxicação Metabólica Fase I/genética , Palaemonidae/genética , Palaemonidae/metabolismo , Transcriptoma , Animais , Argentina , Biomarcadores/análiseRESUMO
The effect of the water-soluble fraction (WSF) of crude oil on lipid contents, lipid classes, FA, and PC molecular species was studied in high-phospholipid (hepatopancreas) and low-phospholipid (egg) tissues of a freshwater crustacean. After a 21-d exposure to a sublethal concentration of WSF, a significant decrease in shrimp total lipids was observed, although no alterations could be detected in the hepatopancreas or egg lipid contents. TAG/phospholipid ratios increased in the hepatopancreas and decreased in the eggs, suggesting alterations either in the mobilization of TAG to phospholipid pools or in the energy balance. The FA composition of phosphoglycerides in the hepatopancreas and eggs was dominated by PUFA, whereas the n-3/n-6 ratio was not affected by WSF exposure, although there was a significant increase in hepatopancreas 18:1n-9. Analysis of the PC molecular species by HPLC-ELSD showed the presence of 15 species, with 16:0/18:1, 18:1/18:2, 16:0/20:5, and 16:1/20:5 being the major species in the hepatopancreas. The PC molecular species in the eggs showed a different pattern, dominated by 16:0/18:1 and 18:1/18:2. Of the PC molecular species, 10 contained 22:6n-3, 20:5n-3, and 20:4n-6. Small amounts of di-PUFA species were also found. Exposure to WSF altered the PC molecular species in both tissues. The four major hepatopancreas molecular species and most of the ones containing PUFA decreased. This was compensated for by an increase in 16:1/18:1 (152%) and 18:1/18:1 (50%). The two major egg PC molecular species decreased, whereas the PUFA-containing ones increased. The contrasting responses of both tissues to WSF contamination suggests the presence of different homeostatic mechanisms.
Assuntos
Palaemonidae/efeitos dos fármacos , Petróleo/efeitos adversos , Fosfatidilcolinas/química , Poluentes Químicos da Água/farmacologia , Animais , Cromatografia Líquida de Alta Pressão , Ácidos Graxos/análise , Feminino , Hepatopâncreas/química , Hepatopâncreas/efeitos dos fármacos , Óvulo/química , Óvulo/efeitos dos fármacos , Palaemonidae/metabolismo , SolubilidadeRESUMO
Hemocyanin of the spider Polybetes pythagoricus, in addition to its typical role as an oxygen transporter, also exhibits a phenoloxidase activity induced by micellar concentrations of SDS. In the present work, we found the kinetic parameters Km and Vmax of Polybetes pythagoricus hemocyanin (PpHc) PO activity to be 0.407 mM and 0.081 µmolmin(-1) mg protein(-1) , respectively. Dopamine was used as the substrate with SDS at a final concentration of 10 mM and a 30-min incubation at 25°C. Conformational changes in Hc associated with the SDS treatment were analyzed using far-UV circular dichroism, intrinsic fluorescence and absorption spectroscopy. The secondary and tertiary structural changes of PpHc induced by SDS led to increases in α-helical content and tryptophan fluorescence intensity. A reduction in the absorption spectrum at 340 nm in the presence of SDS was also observed. These results suggest that the SDS-induced PO activity of PpHc can be ascribed to conformational changes in the local environment of the typer-3 copper active site.
Assuntos
Hemocianinas/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Aranhas/enzimologia , Animais , CinéticaRESUMO
BACKGROUND: The chemical control of the Chagas disease vector Triatoma infestans is endangered by the emergence of pyrethroid resistance. An effective alternative control tool is the use of the entomopathogenic fungus Beauveria bassiana. The effect of deltamethrin on fungal growth, gene expression and enzyme activity in relation to detoxification, antioxidant response and oxidative stress levels was studied to evaluate fungal tolerance to deltamethrin. RESULTS: The mean inhibitory concentration (IC50 ) was 50 µg deltamethrin/cm(2). Cytochrome P450 genes were differentially expressed; cyp52X1 and cyp617N1 transcripts were > 2-fold induced, followed by cyp655C1 (1.8-fold). Minor effects were observed on genes encoding for other P450s, epoxide hydrolase and glutathione S-transferase (GST). Superoxide dismutase (SOD) genes showed induction levels ≤ 2, catalase (CAT) and glutathione peroxidase genes were also induced â¼ 2-3-fold and < 2-fold, respectively. The activities of enzymes participating in the antioxidant defense system and phase II detoxification were also evaluated; SOD, CAT and GST activity showed significant differences with deltamethrin concentration. Lipid peroxidation levels and free proline content were also altered. CONCLUSIONS: Beauveria bassiana GHA can be used combined with deltamethrin without significant metabolic detrimental effects. This combination will help optimizing the benefits and increasing the efficacy of vector control tools.
Assuntos
Beauveria/efeitos dos fármacos , Beauveria/fisiologia , Inseticidas/toxicidade , Nitrilas/toxicidade , Piretrinas/toxicidade , Animais , Antioxidantes/metabolismo , Beauveria/genética , Agentes de Controle Biológico , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Expressão Gênica/efeitos dos fármacos , Concentração Inibidora 50 , Peroxidação de Lipídeos/efeitos dos fármacos , Prolina/metabolismo , TriatomaRESUMO
The shuttle system that mediates the transport of fatty acids across the mitochondrial membrane in invertebrates has received little attention. Carnitine O-palmitoyltransferase I (EC 2.3.1.21; CPT I) is a key component of this system that in vertebrates controls long-chain fatty acid beta-oxidation. To gain knowledge on the acyltransferases in aquatic arthropods, physical, kinetic, regulatory and immunological properties of CPT of the midgut gland mitochondria of Macrobrachium borellii were assayed. CPT I optimum conditions were 34 degrees C and pH=8.0. Kinetic analysis revealed a Km for carnitine of 2180+/-281 microM and a Km for palmitoyl-CoA of 98.9+/-8.9 microM, while V(max) were 56.5+/-6.6 and 36.7+/-4.8 nmol min(-1) mg protein(-1), respectively. A Hill coefficient, n~1, indicate a Michaelis-Menten behavior. The CPT I activity was sensitive to regulation by malonyl-CoA, with an IC(50) of 25.2 microM. Electrophoretic and immunological analyses showed that a 66 kDa protein with an isoelectric point of 5.1 cross-reacted with both rat liver and muscle-liver anti CPT I polyclonal antibodies, suggesting antigenic similarity with the rat enzymes. Although CPT I displayed kinetic differences with insect and vertebrates, prawn showed a high capacity for energy generation through beta-oxidation of long-chain fatty acids.