Hydrogen peroxide and chlorine dioxide against parasite Ichthyophthirius multifiliis (Protozoa, Ciliophora) in jundiá fingerlings

ABSTRACT: Ichthyophthiriasis is a worldwide fish disease with great financial impact on freshwater fish farming due to its associated high mortality rates. Current study assesses the parasiticidal capacity of hydrogen peroxide (H2O2) and chlorine dioxide (ClO2) against the causative agent, Ichthyophthirius multifiliis, in jundiá. Median lethal concentration (LC50, 96h) of each chemical agent was established, as well as the minimum inhibitory concentration of hydrogen peroxide for the parasite´s infectious larval phase (theront). Products were tested asynchronously in parasitized fingerlings for short and long baths at the following concentrations and exposure times: 1. Hydrogen peroxide: (T1) continuous bath - 30ppm and (T2) 50ppm; (T3) short bath - 150ppm, during 1h and (T4) 250ppm during 1h; control group (without any chemical agent). 2. Chlorine dioxide: (T1) continuous bath - 4ppm and (T2) 20ppm; (T3) short bath - 200ppm, during 1min; (T4) short bath - 400ppm, during 1min and control group. Data analysis demonstrated a concentration of 82.54ppm of the commercial product (or 24.76ppm of the active chemical agent) as LC50, 96h of H2O2 and 38.4ppm product (or 2.68ppm of the active chemical agent) for ClO2. Hydrogen peroxide concentration causing 100% mortality rate of theronts in 1h was 25ppm (product, or 7.5ppm of the active chemical agent). At the end of the fourth day of curative experiment, 98% of the animals died by ichthyophthiriasis. No treatment was effective against the parasite.

Dopamine Activates Two Different Receptors to Produce Variability in Sign at an Identified Synapse

Dopamine activates two different receptors to produce variability in sign at an identified synapse. Chemical synaptic transmission was investigated at a central synapse between identified neurons in the freshwater snail, Lymnaea stagnalis. The presynaptic neuron was the dopaminergic cell, Right Pedal Dorsal one (RPeD1). The postsynaptic neuron was Visceral Dorsal four (VD4). These neurons are components of the respiratory central pattern generator. The synapse from RPeD1 to VD4 showed variability of sign, i.e., it was either inhibitory (monophasic and hyperpolarizing), biphasic (depolarizing followed by hyperpolarizing phases), or undetectable. Both the inhibitory and biphasic synapse were eliminated by low Ca2+/high Mg2+ saline and maintained in high Ca2+/high Mg2+ saline, indicating that these two types of connections were chemical and monosynaptic. The latency of the inhibitory postsynaptic potential (IPSP) in high Ca2+/high Mg2+ saline was ∼43 ms, whereas the biphasic postsynaptic potential (BPSP) had ∼12-ms latency in either normal or high Ca2+/high Mg2+ saline. For a given preparation, when dopamine was pressured applied to the soma of VD4, it always elicited the same response as the synaptic input from RPeD1. Thus, for a VD4 neuron receiving an IPSP from RPeD1, pressure application of dopamine to the soma of VD4 produced an inhibitory response similar to the IPSP. The reversal potentials of the IPSP and the inhibitory dopamine response were both approximately −90 mV. For a VD4 neuron with a biphasic input from RPeD1, pressure-applied dopamine produced a biphasic response similar to the BPSP. The reversal potentials of the depolarizing phase of the BPSP and the biphasic dopamine response were both approximately −44 mV, whereas the reversal potentials for the hyperpolarizing phases were both approximately −90 mV. The hyperpolarizing but not the depolarizing phase of the BPSP and the biphasic dopamine response was blocked by the d-2 dopaminergic antagonist (±) sulpiride. Previously, our laboratory demonstrated that both IPSP and the inhibitory dopamine response are blocked by (±) sulpiride. Conversely, the depolarizing phase of both the BPSP and the biphasic dopamine response was blocked by the Cl− channel antagonist picrotoxin. Finally, both phases of the BPSP and the biphasic dopamine response were desensitized by continuous bath application of dopamine. These results indicate that the biphasic RPeD1 → VD4 synapse is dopaminergic. Collectively, these data suggest that the variability in sign (inhibitory vs. biphasic) at the RPeD1 → VD4 synapse is due to activation of two different dopamine receptors on the postsynaptic neuron VD4. This demonstrates that two populations of receptors can produce two different forms of transmission, i.e., the inhibitory and biphasic forms of the single RPeD1 → VD4 synapse.

GTP gammaS removal of D-600 block of skeletal muscle excitation-contraction coupling

G proteins interacting with dihydropyridine receptors (DHPR) in transverse tubules (TT) of skeletal muscle may have a role in skeletal excitation-contraction (EC) coupling. The aim of this study was to determine the effects of G protein-specific nucleotides [guanosine 5'-O-(3-thiotriphosphate) (GTP gammaS) and guanosine 5'-O-(2-thiodiphosphate) (GDP betaS)] on the EC coupling mechanism in the presence of D-600, an agent that blocks EC coupling by immobilizing the voltage-sensing subunit of the DHPR in its inactivated state. By use of the mechanically peeled single-fiber preparation from rabbit adductor magnus skeletal muscle, 50 microM GTP gammaS and 500 microM GDP betaS were applied with the fiber in a D-600-induced state of blocked EC coupling. Neither nucleotide served as an independent stimulus for sarcoplasmic reticulum (SR) Ca2+ release when added to the TT polarizing bath under conditions of D-600 block. The presence of GTP gammaS or GDP betaS during a complete EC coupling cycle removed the D-600 block of EC coupling, despite continuous bath D-600. After the nucleotides were washed out, in the continued presence of D-600, the D-600 block of EC coupling was reestablished. In contrast, GTP gammaS added only during the period of TT depolarization under D-600 block did not remove the D-600 block of EC coupling, even though GTP gammaS did stimulate SR Ca2+ release. GTP gammaS had no effect on submaximum (0.5-1.0 mM) caffeine contractures and thus is unlikely to be acting through the Ca2+-induced Ca2+ release mechanism of the SR. These data suggest that the molecular binding site for GTP gammaS and GDP betaS is likely to be in the TT near the DHPR, perhaps on a G protein.