The Insecticidal Activity of Rhinella schneideri (Werner, 1894) Paratoid Secretion in Nauphoeta cinerea Cocroaches.

Rhinella schneideri is a common toad found in South America, whose paratoid toxic secretion has never been explored as an insecticide. In order to evaluate its insecticidal potential, Nauphoeta cinerea cockroaches were used as an experimental model in biochemical, physiological and behavioral procedures. Lethality assays with Rhinella schneideri paratoid secretion (RSPS) determined the LD50 value after 24 h (58.07µg/g) and 48 h exposure (44.07 µg/g) (R2 = 0.882 and 0.954, respectively). Acetylcholinesterase activity (AChE) after RSPS at its highest dose promoted an enzyme inhibition of 40%, a similar effect observed with neostigmine administration (p < 0.001, n= 5). Insect locomotion recordings revealed that RSPS decreased the distance traveled by up to 37% with a concomitant 85% increase in immobile episodes (p < 0.001, n = 36). RSPS added to in vivo cockroach semi-isolated heart preparation promoted an irreversible and dose dependent decrease in heart rate, showing a complete failure after 30 min recording (p < 0.001, n ≥ 6). In addition, RSPS into nerve-muscle preparations induced a dose-dependent neuromuscular blockade, reaching a total blockage at 70 min at the highest dose applied (p < 0.001, n ≥ 6). The effect of RSPS on spontaneous sensorial action potentials was characterized by an increase in the number of spikes 61% (p < 0.01). Meanwhile, there was 42% decrease in the mean area of those potentials (p < 0.05, n ≥ 6). The results obtained here highlight the potential insecticidal relevance of RSPS and its potential biotechnological application.

Modulation of octopaminergic and cholinergic pathways induced by Caatinga tree Manilkara rufula chemical compounds in Nauphoeta cinerea cockroaches.

The entomotoxic potential of Manilkara rufula crude extract (CEMR) and its aqueous (AFMR) and methanolic (MFMR) fractions were evaluated against Nauphoeta cinerea cockroaches. The results point out to a direct modulation of octopaminergic and cholinergic pathways in insect nervous system. CEMR induced an anti-acetylcholinesterase (AChE) effect in cockroach brain homogenates. CEMR significantly decreased the cockroach heart rate in semi-isolated heart preparations. CEMR also caused a broad disturbance in the insect behavior by reducing the exploratory activity. The decreased antennae and leg grooming activities, by different doses of CEMR, mimicked those of phentolamine activity, a selective octopaminergic receptor antagonist. The lethargy induced by CEMR was accompanied by neuromuscular failure and by a decrease of sensilla spontaneous neural compound action potentials (SNCAP) firing in in vivo and ex vivo cockroach muscle-nerve preparations, respectively. AFMR was more effective in promoting neuromuscular paralysis than its methanolic counterpart, in the same dose. These data validate the entomotoxic activity of M. rufula. The phentolamine-like modulation induced in cockroaches is the result of a potential direct inhibition of octopaminergic receptors, combined to an anti-AChE activity. In addition, the modulation of CEMR on octopaminergic and cholinergic pathways is probably the result of a synergism between AFMR and MFMR chemical compounds. Further phytochemical investigation followed by a bio-guiding protocol will improve the molecular aspects of M. rufula pharmacology and toxicology to insects.