Fluorescence enhancing and quenching signal based on new approach for selective detection of multiple organochlorine pesticides using blue emissive-carbon dot.

Measuring the concentration of organochlorine pesticides (OCPs) in agriculture has engaged significant awareness for healthcare investigation since OCPs are harmful to many physiological processes. Excessive usage of these compounds can result in major contamination of the environment and food supply chains. As a result, more accurate and rapid ways to detect pesticide residues in food are required. In this work, we have portrayed the surface-engineered fluorescent blue emissive-carbon dot (B-CD) with a high quantum yield (49.3 %) via the hydrothermal method for fluorescent sensing of OCPs on real samples. The amine group functionalities of carbon dots have supported the direct coordination with -Cl and -OH groups of HEP, ENS, CDF and 2,4-DPAC for the sensitive detection of OCPs, by switching in the fluorescent intensity of B-CD. The functional group of OCPs exhibits a variety of binding interactions with B-CD to contribute a complex formation, which leads to static quenching via an insubstantial restricted electron transfer process. The synthesized carbon dots exhibit individuality in binding nature towards different OCPs. Fluorescence studies help to distinguish the target OCPs and their low detection limits (LODs) were 0.002, 0.099, 0.16 and 0.082 µM for Heptachlor (HEP - turn "on"), Endosulfan (ENS), Chlordimeform (CDF) and 2,4-dichlorophenoxyacetic acid (2,4-DPAC - turn "off") OCPs respectively. The real water samples and agriculture food samples were effectively investigated and the OCP toxicity was noted. Thus, the design of the fluorescence sensor is established as an easy and proficient sensing method for detecting OCPs.

Rapid and sensitive detection of chlordimeform in cucumber and tomato samples using an immunochromatographic assay.

Chlordimeform (CDM) is a broad-spectrum and highly effective insecticide and acaricide used to control pests in agriculture. We produced two monoclonal antibodies (mAbs) against CDM and developed an immunochromatographic assay to screen CDM in cucumbers and tomatoes. MAb 4A3 had high sensitivity with a 50% inhibitory concentration of 0.287 ng mL-1. The assay had a cut-off value of 25 µg kg-1 and a visual limit of detection (vLOD) of 1 µg kg-1 in cucumbers and a cut off value of 50 µg kg-1 and a vLOD of 2.5 µg kg-1 in tomatoes. The calculated limit of detection (cLOD) in cucumbers and tomatoes was 0.115 µg kg-1 and 0.215 µg kg-1, respectively. The recovery rates were 97.9% to 106.9% for cucumbers and 97.8% to 107.4% for tomatoes, consistent with the results obtained from indirect competitive ELISA. Our findings showed that the immunochromatographic assay is an efficient and accurate method for CDM detection in cucumbers and tomatoes.

The preparation of polyclonal antibody against chlordimeform and establishment of detection by indirect competitive ELISA.

Objective: Chlordimeform is a chemical pesticide that is highly carcinogenic and toxic. The purpose of this study was to establish an enzyme-linked immunosorbent assay (ELISA) method for the detection of chlordimeform in aquaculture and fish farming. METHODS: Chlordimeform was coupled with bovine serum albumin (BSA) and ovalbumin (OVA) as carrier proteins. A chlordimeform-BSA conjugate was used as an immunogen, and chlordimeform-OVA was used as a coating antigen. Chlordimeform-BSA was used to immunize rabbits, and a polyclonal antibody was prepared. An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was established to detect chlordimeform. RESULTS: The working range of the established IC-ELISA method for chlordimeform detection was 1-20 ng/mL. The IC50 was 3.126 ng/mL, and the lower limit of detection (LOD) of chlordimeform was 0.637 ng/mL. The recovery of chlordimeform from spiked water samples ranged from 81% to 107%. CONCLUSION: An anti-chlordimeform polyclonal antibody was successfully developed, and a novel IC-ELISA was established to detect chlordimeform in aquaculture.

The physiological basis for contrast opponency in motion computation in Drosophila.

In Drosophila, direction-selective neurons implement a mechanism of motion computation similar to cortical neurons, using contrast-opponent receptive fields with ON and OFF subfields. It is not clear how the presynaptic circuitry of direction-selective neurons in the OFF pathway supports this computation if all major inputs are OFF-rectified neurons. Here, we reveal the biological substrate for motion computation in the OFF pathway. Three interneurons, Tm2, Tm9 and CT1, provide information about ON stimuli to the OFF direction-selective neuron T5 across its receptive field, supporting a contrast-opponent receptive field organization. Consistent with its prominent role in motion detection, variability in Tm9 receptive field properties transfers to T5, and calcium decrements in Tm9 in response to ON stimuli persist across behavioral states, while spatial tuning is sharpened by active behavior. Together, our work shows how a key neuronal computation is implemented by its constituent neuronal circuit elements to ensure direction selectivity.

Octopaminergic system orchestrates combat and mating behaviors: A potential regulator of alternative male mating tactics in an armed beetle.

Male-male combats over females and territories are widespread across animal taxa. The winner of a combat gains resources, while the loser suffers significant costs (e.g. time, energy and injury) without gaining resources. Many animals have evolved behavioral flexibility, depending on their nutritional condition and experience, to avoid combat in order to reduce such costs. In these cases, male aggression often correlates with mating behavior changes, that is, the deployment of alternative reproductive tactics. Therefore, uncovering the physiological mechanism that orchestrates combat and mating behaviors is essential to understand the evolution of alternative mating tactics. However, so far, our knowledge is limited to specific behaviors (i.e., fighting or mating) of specific model species. In this study, we used an armed beetle (Gnatocerus cornutus) and hypothesized that one of the key neuromodulators of invertebrate aggression, octopamine (OA), would control male combat and other mating behaviors. Using receptor agonists (chlordimeform and benzimidazole), we showed that the octopaminergic (OAergic) system down-regulated the combat and courtship behaviors, while it up-regulated locomotor activity and sperm size. This suggests that the OAergic system orchestrates a suite of fighting and mating behaviors, thereby implying that correlated behavioral responses to OAergic signaling may have driven the evolution of alternative mating tactics in this beetle.

ZnFe2O4-chitosan magnetic beads for the removal of chlordimeform by photo-Fenton process under UVC irradiation.

A simple protocol was proposed for the preparation of magnetic chitosan beads ZnFe2O4-CS via a co-precipitation method. The use of synthesized magnetic ZnFe2O4-CS beads as catalyst for the heterogeneous photo-Fenton treatment of chlordimeform insecticide (CDM) was evaluated. The photo-Fenton experiments were carried out with different synthesized catalysts by varying the molar ratio Zn/Fe in chitosan beads, the catalyst concentration and pH. Under optimal conditions using 1 g of ZnFe2O4-CS beads with a molar ratio Zn/Fe = 0.35 and at pHinitial = 3, a real wastewater doped with 20 mg L-1 of CDM was treated and complete removal of the insecticide was achieved after 7 min with a total TOC removal after 2 h of treatment. The generated carboxylic acids and ions during the photo-Fenton process were identified and quantified. The stability of the photocatalytic activity of the best catalyst in terms of pollutant removal, ZnFe2O4-CS(0.35) beads with a molar ratio Zn/Fe equal to 0.35, was satisfactory validated by four consecutive cycles. This optimal catalyst was characterized, before and after use, by Scanning Electron Microscopy/Energy Dispersive X-Ray Spectroscopy, X-Ray Powder Diffraction and Vibrating Sample Magnetometry analysis.

Toxicological Evaluation of Novel Butenolide Pesticide Flupyradifurone Against Culex quinquefasciatus (Diptera: Culicidae) Mosquitoes.

The impact of increasing resistance of mosquitoes to conventional pesticides has led to investigate various unique tools and pest control strategies. Herein, we assessed the potency of flupyradifurone, a novel pesticide, on fourth instar larvae of Culex quinquefasciatus Say. Further, we evaluated the synergistic action of piperonyl butoxide (PBO) and the octopamine receptor agonists (OR agonists) chlordimeform (CDM) and amitraz (AMZ) on the toxicity of flupyradifurone in comparison with sulfoxaflor and nitenpyram to increase their toxicity on Cx. quinquefasciatus. Results demonstrated that flupyradifurone was the most potent pesticide followed by sulfoxaflor and nitenpyram. Further, the synergetic effect of PBO, CDM, and AMZ was significant for all selected pesticides especially flupyradifurone. However, AMZ had the most significant effect in combination with the selected pesticides followed by CDM and PBO. The toxicity of the pesticides was time-dependent and increased over time from 24, 48, to 72 h of exposure in all experiments. The results indicate that flupyradifurone is a promising component in future mosquito control programs.

Pre-adult aggression and its long-term behavioural consequences in crickets.

Social experience, particularly aggression, is considered a major determinant of consistent inter-individual behavioural differences between animals of the same species and sex. We investigated the influence of pre-adult aggressive experience on future behaviour in male, last instar nymphs of the cricket Gryllus bimaculatus. We found that aggressive interactions between male nymphs are far less fierce than for adults in terms of duration and escalation. This appears to reflect immaturity of the sensory apparatus for releasing aggression, rather than the motor system controlling it. First, a comparison of the behavioural responses of nymphs and adults to mechanical antennal stimulation using freshly excised, untreated and hexane-washed antennae taken from nymphs and adults, indicate that nymphs neither respond to nor produce sex-specific cuticular semiochemicals important for releasing aggressive behaviour in adults. Second, treatment with the octopamine agonist chlordimeform could at least partially compensate for this deficit. In further contrast to adults, which become hyper-aggressive after victory, but submissive after defeat, such winner and loser effects are not apparent in nymphs. Aggressive competition between nymphs thus appears to have no consequence for future behaviour in crickets. Male nymphs are often attacked by adult males, but not by adult females. Furthermore, observations of nymphs raised in the presence, or absence of adult males, revealed that social subjugation by adult males leads to reduced aggressiveness and depressed exploratory behaviour when the nymphs become adult. We conclude that social subjugation by adults during pre-adult development of nymphs is a major determinant of consistent inter-individual behavioural differences in adult crickets.

Simultaneous determination of amitraz, chlordimeform, formetanate and metabolites in human blood by liquid chromatography tandem mass spectrometry with phospholipid-removal pretreatment.

In this study, a reliable ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method coupled with an easy, fast and effective sample pretreatment procedure was developed for simultaneous determination of amitraz, chlordimeform, formetanate and their metabolites in human blood. With the procedures of protein precipitation and a phospholipid-removal step, the endogenous compound interference was significantly reduced, and matrix effects were significantly reduced. The linear ranges of matrix-matched standard curves were from 0.5 to 1000 ng/mL with coefficients of determination >0.996. Very low limits of detection (0.05-0.12 ng/mL) and limits of quantitation (0.15-0.4 ng/mL) were achieved. Reasonable recoveries ranging from 88.1 to 103.5% were obtained. The intra-day RSDs ranging from 3.2 to 8.6% and inter-day RSDs ranging from 4.8 to 9.2% indicated good precision. With the introduction of a phospholipid-removal step, the ME ranged from 90.1 to 98.5%. The established method was successfully applied to the analysis of a blood sample from a formetanate poisoning case. This method possesses the advantages of high sensitivity, reduced matrix effects and rapidity.

Simultaneous determination of amitraz, chlordimeform, formetanate and their main metabolites in human urine by high performance liquid chromatography-tandem mass spectrometry.

A rapid, simple and reliable high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for simultaneous determination of amitraz, chlordimeform, formetanate and their main metabolites, N-(2,4-dimethylphenyl)-N-methyl-formamidine (DMPF), 2,4-dimethylformamidine (DMF), 2,4-dimethylaniline (DMA), 4-chloro-2-methylaniline and 3-hydroxyacetanilide in human urine. The urine samples were mixed with buffer solutions (pH 8) and subsequently cleaned up by solid supported liquid/liquid extraction (SLE). The target analytes were efficiently separated with a Waters Atlantis T3 column (150mm×4.6mm, 5µm), ionized with electrospray ion source in positive mode, and quantitatively determined by tandem mass spectrometry in the multiple reaction monitoring (MRM) mode. In order to minimize matrix effects, the matrix-matched calibration curves of eight analytes were adopted with correlation coefficients (R2) above 0.99. The method were further validated by determining the limits of detection (LODs, 0.3-0.6ng/mL), the limits of quantitation (LOQs, 1.0-2.0ng/mL) and recoveries (89.1%-108.4%) with intra-day and inter-day relative standard deviation (RSD, <11%). The established method was applied and demonstrated in a real case by assaying a urine sample from a female poisoned by formetanate. The achieved results proved this method to be rapid, sensitive and accurate for simultaneous quantitation of eight analytes in human urine for intended forensic cases of human poisoning.

The role of octopamine receptor agonists in the synergistic toxicity of certain insect growth regulators (IGRs) in controlling Dengue vector Aedes aegypti (Diptera: Culicidae) mosquito.

The synergistic action of octopamine receptor agonists (OR agonists) on many insecticide classes (e.g., organophosphorus, pyrethroids, and neonicotinoids) on Aedes aegypti L. has been reported recently. An investigation of OR agonist's effect on insect growth regulators (IGRs) was undertaken to provide a better understanding of the mechanism of action. Based on the IGR bioassay, pyriproxyfen was the most potent IGR insecticide tested (EC50=0.0019ng/ml). However, the lethal toxicity results indicate that diafenthiuron was the most potent insecticide (LC50=56ng/cm(2)) on A. aegypti adults after 24h of exposure. The same trend was true after 48 and 72h of exposure. Further, the synergistic effects of OR agonists plus amitraz (AMZ) or chlordimeform (CDM) was significant on adults. Among the tested synergists, AMZ increased the potency of the selected IGRs on adults the greatest. As results, OR agonists were largely synergistic with the selected IGRs. OR agonists enhanced the lethal toxicity of IGRs, which is a valuable new tool in the field of A. aegypti control. However, further field experiments need to be done to understand the unique potential role of OR agonists and their synergistic action on IGRs.

Synergistic action of octopamine receptor agonists on the activity of selected novel insecticides for control of dengue vector Aedes aegypti (Diptera: Culicidae) mosquito.

Studying insecticide resistance in mosquitoes has attracted the attention of many scientists to elucidate the pathways of resistance development and to design novel strategies in order to prevent or minimize the spread and evolution of resistance. Here, we tested the synergistic action of piperonyl butoxide (PBO) and two octopamine receptor (OR) agonists, amitraz (AMZ) and chlordimeform (CDM) on selected novel insecticides to increase their lethal action on the fourth instar larvae of Aedes aegypti L. However, chlorfenapyr was the most toxic insecticide (LC50 = 193, 102, and 48 ng/ml, after 24, 48, and 72 h exposure, respectively) tested. Further, PBO synergized all insecticides and the most toxic combinatorial insecticide was nitenpyram even after 48 and 72 h exposure. In addition, OR agonists significantly synergized most of the selected insecticides especially after 48 and 72 h exposure. The results imply that the synergistic effects of amitraz are a promising approach in increasing the potency of certain insecticides in controlling the dengue vector Ae. aegypti mosquito.

Unique biochemical and molecular biological mechanism of synergistic actions of formamidine compounds on selected pyrethroid and neonicotinoid insecticides on the fourth instar larvae of Aedes aegypti (Diptera: Culicidae).

We recently reported that formamidine pesticides such as amitraz and chlordimeform effectively synergize toxic actions of certain pyrethroid and neonicotinoid insecticides in some insect species on the 4th instar larvae of Aedes aegypti. Here we studied the biochemical basis of the synergistic actions of the formamidines in amplifying the toxicity of neonicotinoids and pyrethroids such as dinotefuran and thiamethoxam, as well as deltamethrin-fenvalerate type of pyrethroids. We tested the hypothesis that their synergistic actions are mediated by the octopamine receptor, and that the major consequence of octopamine receptor activation is induction of trehalase to increase glucose levels in the hemolymph. The results show that formamidines cause a significant up-regulation of the octopamine receptor and trehalase mRNA expressions. Furthermore, formamidines significantly elevate levels of free glucose when co-treated with dinotefuran, deltamethrin and fenvalerate, but not with permethrin or fenitrothion, which showed no synergistic toxic effects with formamidines. These results support the conclusion that the main mode of synergism is based on the ability to activate the octopamine receptor, which is particularly effective with insecticides causing hyperexcitation-induced glucose release and consequently leading to quick energy exhaustion.

Synergistic actions of formamidine insecticides on the activity of pyrethroids and neonicotinoids against Aedes aegypti (Diptera: Culicidae).

Formamidines are unique insecticides and acaricides that elicit multiple effects in controlling insects. Here, we tested two formamidines, amitraz, and chlordimeform, for their synergistic actions on type II pyrethroids and neonicotinoids to increase their larvicidal actions on the fourth instars of Aedes aegypti L. An organophosphate insecticide was used as a negative control. After 24 h, the synergism of formamidines was highest on imidacloprid, followed by two type II pyrethroids, deltamethrin and fenvalerate. After 48 h, the synergism of formamidines on imidacloprid decreased, remained unchanged on type II pyrethroids, and increased noticeably on two of the newer type neonicotinoids, dinotefuran and thiamethoxam. By 72 h, synergism of formamidines on dinotefuran reached the maximum, while that on imidacloprid was at a minimum. Both formamidines did not show synergistic effects on permethrin or fenitrothion. In all cases, the synergistic effects of amitraz on the two major classes of larvicides were greater than for chlordimeform. These results indicate that amitraz is a promising synergist that shows the potential to increase the efficacy of certain members of type II pyrethroids as well as neonicotinoids to control Ae. aegypti larvae.

[Effect of scopolamine on the acute severe chlorphenamidine poisoning patients].

OBJECTIVE: To investigate effectiveness of scopolamine on the acute severe chlorphenamidine poisoning patients. METHODS: 72 cases of acute severe chlorphenamidine poisoning patients were divided into I and II groups by the principle of a 1:1 sampling according to the order of admission. The I group (36 cases) were treated with traditional multimodality therapy, including gastrolavage, catharsis, using reductant-oxidant (methylthioninium chloride and vitamin C), and symptomatic treatment. The II group were treated with traditional multimodality therapy and scopolamine at the same times. Blood methemoglobin were measured at 0, third, seventh, twelfth, twenty-fourth hour, serum troponin I (CTnI) and creatine kinase isoenzyme (CK-MB) levels at third, seventh, twenty-fourth, forty-eighth hour, hepatic and renal functions at third, twenty-fourth, forty-eighth hour, and electrocardiogram (ECG) were evaluated every 4 hours in 3 days after hospitalization on all patients. The two groups of patients were compared the efficacy and change detection of targets. RESULTS: 31 patients (86.11%) recovered and 5 patients (13.89%) died in I group. All 36 cases recovered in II group. The recovery rate of II group was distinctively higher than that in I group (P < 0.05) and the difference was statistically significant (P < 0.05). The average recovery time and the length of hospital stay in II group were sharply shorter than those in I group (P < 0.01) and the difference was statistically significant (P < 0.05). Serum CTnI levels between seventh hour and forty-eighth hour, serum CK-MB levels between third hour and forty-eighth hour and methemoglobin concentration at third, seventh, twelfth, twenty-fourth hour were apparently lower in II group, and the difference was statistically significant (P < 0.05). The abnormal rates of hepatic and renal functions in II group were distinctively lower than those in I group and the difference was statistically significant (P < 0.05). The abnormal rates of ECG in the second and third day in II group were respectively 38.89% and 11.11%, and were lower than those in I group (64.71%, 38.71%). The difference was statistically significant (P < 0.05). CONCLUSION: Scopolamine has the excellent treatment effect on acute severe chlorphenamidine poisoning patients and protec their hearts, livers, and kidneys. It complements the deficiency of reductant-oxidants, and combination of the two drugs can form the synergy effect.

Octopaminergic modulation of contrast gain adaptation in fly visual motion-sensitive neurons.

Locomotor activity like walking or flying has recently been shown to alter visual processing in several species. In insects, the neuromodulator octopamine is thought to play an important role in mediating state changes during locomotion of the animal [K.D. Longden & H.G. Krapp (2009) J. Neurophysiol., 102, 3606-3618; (2010) Front. Syst. Neurosci., 4, 153; S.N. Jung et al. (2011)J. Neurosci., 31, 9231-9237]. Here, we used the octopamine agonist chlordimeform (CDM) to mimic effects of behavioural state changes on visual motion processing. We recorded from identified motion-sensitive visual interneurons in the lobula plate of the blowfly Calliphora vicina. In these neurons, which are thought to be involved in visual guidance of locomotion, motion adaptation leads to a prominent attenuation of contrast sensitivity. Following CDM application, the neurons maintained high contrast sensitivity in the adapted state. This modulation of contrast gain adaptation was independent of the activity of the recorded neurons, because it was also present after stimulation with visual motion that did not result in deviations from the neurons' resting activity. We conclude that CDM affects presynaptic inputs of the recorded neurons. Accordingly, the effect of CDM was weak when adapting and test stimuli were presented in different parts of the receptive field, stimulating separate populations of local presynaptic neurons. In the peripheral visual system adaptation depends on the temporal frequency of the stimulus pattern and is therefore related to pattern velocity. Contrast gain adaptation could therefore be the basis for a shift in the velocity tuning that was previously suggested to contribute to state-dependent processing of visual motion information in the lobula plate interneurons.

Effect of scopolamine on the acute severe chlorphenamidine poisoning patients / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate effectiveness of scopolamine on the acute severe chlorphenamidine poisoning patients.</p><p><b>METHODS</b>72 cases of acute severe chlorphenamidine poisoning patients were divided into I and II groups by the principle of a 1:1 sampling according to the order of admission. The I group (36 cases) were treated with traditional multimodality therapy, including gastrolavage, catharsis, using reductant-oxidant (methylthioninium chloride and vitamin C), and symptomatic treatment. The II group were treated with traditional multimodality therapy and scopolamine at the same times. Blood methemoglobin were measured at 0, third, seventh, twelfth, twenty-fourth hour, serum troponin I (CTnI) and creatine kinase isoenzyme (CK-MB) levels at third, seventh, twenty-fourth, forty-eighth hour, hepatic and renal functions at third, twenty-fourth, forty-eighth hour, and electrocardiogram (ECG) were evaluated every 4 hours in 3 days after hospitalization on all patients. The two groups of patients were compared the efficacy and change detection of targets.</p><p><b>RESULTS</b>31 patients (86.11%) recovered and 5 patients (13.89%) died in I group. All 36 cases recovered in II group. The recovery rate of II group was distinctively higher than that in I group (P < 0.05) and the difference was statistically significant (P < 0.05). The average recovery time and the length of hospital stay in II group were sharply shorter than those in I group (P < 0.01) and the difference was statistically significant (P < 0.05). Serum CTnI levels between seventh hour and forty-eighth hour, serum CK-MB levels between third hour and forty-eighth hour and methemoglobin concentration at third, seventh, twelfth, twenty-fourth hour were apparently lower in II group, and the difference was statistically significant (P < 0.05). The abnormal rates of hepatic and renal functions in II group were distinctively lower than those in I group and the difference was statistically significant (P < 0.05). The abnormal rates of ECG in the second and third day in II group were respectively 38.89% and 11.11%, and were lower than those in I group (64.71%, 38.71%). The difference was statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>Scopolamine has the excellent treatment effect on acute severe chlorphenamidine poisoning patients and protec their hearts, livers, and kidneys. It complements the deficiency of reductant-oxidants, and combination of the two drugs can form the synergy effect.</p>

The effect of octopaminergic compounds on the behaviour and transmission of Gyrodactylus.

BACKGROUND: The high transmission potential of species belonging to the monogenean parasite genus Gyrodactylus, coupled with their high fecundity, allows them to rapidly colonise new hosts and to increase in number. One gyrodactylid, Gyrodactylus salaris Malmberg, 1957, has been responsible for devastation of Altantic salmon (Salmo salar L.) populations in a number of Norwegian rivers. Current methods of eradicating G. salaris from river systems centre around the use of non-specific biocides, such as rotenone and aluminium sulphate.Although transmission routes in gyrodactylids have been studied extensively, the behaviour of individual parasites has received little attention. Specimens of Gyrodactylus gasterostei Gläser, 1974 and G. arcuatus Bychowsky, 1933, were collected from the skin of their host, the three-spined stickleback (Gasterosteus aculeatus L.), and permitted to attach to the substrate. The movements of individual parasites were recorded and analysed. RESULTS: The behaviour patterns of the two species were similar and parasites were more active in red light and darkness than in white light. Four octopaminergic compounds were tested and all four inhibited the movements of parasites. Treatment ultimately led to death at low concentrations (0.2 µM), although prolonged exposure was necessary in some instances. CONCLUSIONS: Octopaminergic compounds may affect the parasite's ability to locate and remain on its host and these or related compounds might provide alternative or supplementary treatments for the control of G. salaris infections. With more research there is potential for use of octopaminergic compounds, which have minimal effects on the host or its environment, as parasite-specific treatments against G. salaris infections.

[Determination of chlordimeform and its metabolite residues in honey using liquid chromatography-tandem mass spectrometry].

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of chlordimeform and its metabolite (4-chloro-o-toluidine) residue in acacia honey, chaste honey, comb honey, multifloral honey and wild honey. The samples were dissolved and diluted with sodium hydroxide solution, and cleaned up by a Waters Oasis HLB solid phase extraction column. The separation was performed on an Agilent XDB-C18 column with gradient elution using 0.1% formic acid solution and acetonitrile. The residues of chlordimeform and its metabolite were determined by electrospray ionization-tandem mass spectrometry, and quantified by the method of matrix-matched external standard. The calibration curves for chlordimeform and its metabolite showed good linearity in the range of 2.5 - 250 microg/L with the correlation coefficients (r) higher than 0.999. The limit of quantification was 5 microg/kg, and the limit of detection was 2.5 microg/kg. The recoveries of chlordimeform and 4-chloro-o-toluidine in honey at the spiked levels of 5, 10 and 20 microg/kg were in the ranges of 75.8% - 113.8% and 85.6% - 114.3%, respectively. The relative standard deviations were 4.8% - 10.2% and 4.7% -9.1%, respectively. The method can meet the requirements for the determination of chlordimeform and 4-chloro-o-toluidine in honey for import and export inspection.

Postembryonic development of centrally generated flight motor patterns in the hawkmoth, Manduca sexta.

This study analyses the maturation of centrally generated flight motor patterns during metamorphosis of Manduca sexta. Bath application of the octopamine agonist chlordimeform to the isolated central nervous system of adult moths reliably induces fictive flight patterns in wing depressor and elevator motoneurons. Pattern maturation is investigated by chlordimeform application at different developmental stages. Chlordimeform also induces motor patterns in larval ganglia, which differ from fictive flight, indicating that in larvae and adults, octopamine affects different networks. First changes in motoneuron activity occur at the pupal stage P10. Rhythmic motor output is induced in depressor, but not in elevator motoneurons at P12. Adult-like fictive flight activity in motoneurons is observed at P16 and increases in speed and precision until emergence 2 days later. Pharmacological block of chloride channels with picrotoxin also induces fictive flight in adults, suggesting that the pattern-generating network can be activated by the removal of inhibition, and that proper network function does not rely on GABA(A) receptors. Our results suggest that the flight pattern-generating network becomes gradually established between P12 and P16, and is further refined until adulthood. These findings are discussed in the context of known physiological and structural CNS development during Manduca metamorphosis.