Nauphoeta Research Articles

RPS6 transcriptional modulation in neural tissues of Nauphoeta cinerea during streptozotocin-associated sugar metabolism impairment.

The use of insects to model molecular events that characterize degenerative conditions was originally met with scepticism. However, the discovery of insect insulin-like peptides in the 1970's and the demonstration of evolutionary conservation of insulin-related signalling from insects to mammals have highlighted the importance and reduced cost of insect models in biomedical research. Here, we expand on our earlier described modelling of streptozotocin-induced brain glucose metabolic disruption in Nauphoeta cinerea, using RNA-sequencing analysis to study the transcriptional and genetic signatures of degeneration and stress signalling when glucose levels are elevated in the brain of the lobster cockroach. Nymphs were randomly divided into three groups: Control (0.8% NaCl), and two single streptozotocin injection doses (74nmol and 740nmol). The transcriptional analyses featured a dysregulation of 226 genes at high dose STZ treatment and 278 genes at the low dose. Our mRNA-sequencing data showed that ribosomal protein genes were the most upregulated genes at both 74 and 740nmol STZ treatment. We therefore used RT-qPCR and relative transcriptional methods to validate our proposed mechanism of brain glucose toxicity-induced degeneration in Nauphoeta cinerea, which involved the upregulation of ribosomal proteins and rpS6 regulators (mTORC1, protein kinases, casein kinase 1 and Death-associated protein kinase), the upregulation of MAPK cascades (RAS, ERK, P38 and JNK), alongside the downregulation of the PI3K/AKT cascade. Taken together, this study highlights the remarkable opportunity for Nauphoeta cinerea use as an experimental organism in hyperglycaemia, degeneration, and stress signalling.

The neurotoxic mechanism of Jack Bean Urease in insects involves the interplay between octopaminergic and dopaminergic pathways

In the last decades, the entomotoxicity of JBU and its derived peptides became an object of study, due mainly to the ubiquitous interaction of these compounds with different species of insects and their potential as natural insecticides. In this work, we investigated the neurotoxic effects of JBU in Nauphoeta cinerea cockroaches by dissecting pharmacologically the monoaminergic pathways involved. Selective pharmacological modulators for monoaminergic pathways in in vivo and ex vivo experimental models were employed. Thus, the analysis of N. cinerea neurolocomotory behavior demonstrated that JBU (1.5 and 3 μg/g) induces a significant decrease in the exploratory activity. In these assays, pretreatment of animals with phentolamine, SCH23390 or reserpine, interfered significantly with the response of JBU. Using in vivo abductor metathoracic preparations JBU (1.5 μg/g) induced progressive neuromuscular blockade, in 120 min recordings. In this set of experiments, the previous treatment of the animals with phentolamine, SCH23390 or reserpine, completely inhibited JBU-induced neuromuscular blockade. The recordings of spontaneous compound neural action potentials in N. cinerea legs showed that JBU, only in the smallest dose, significantly decreased the number of potentials in 60 min recordings. When the animals were pretreated with phentolamine, SCH23390, or reserpine, but not with mianserin, there was a significant prevention of the JBU-inhibitory responses on the action potentials firing. Meanwhile, the treatment of the animals with mianserin did not affect JBU's inhibitory activity. The data presented in this work strongly suggest that the neurotoxic response of JBU in N. cinerea involves a cross talking between OCTOPAMIN-ergic and DOPAMIN-ergic nerve systems, but not the SEROTONIN-ergic neurotransmission. Further molecular biology studies with expression of insect receptors associated with voltage clamp techniques will help to discriminate the selectivity of JBU over the monoaminergic transmission.

A trapdoor spider, Latouchia typica (Araneae: Halonoproctidae), uses vibrational cues as a trigger for predatory behavior

AbstractSpiders are one of the most dominant predators in terrestrial ecosystems. Although cues triggering predatory behavior in web‐building and wandering spiders are well investigated, studies concerning burrowing species, the most ancestral group of spiders, are relatively limited. To clarify critical cues affecting the predatory behavior in burrowing species, we conducted vibration‐reducing experiments with the trapdoor spider Latouchia typica (Araneae: Halonoproctidae) and nymphs of the speckled cockroach, Nauphoeta cinerea (Blattodea: Blaberidae), as prey. Spiders achieved a high success rate of prey capture even when blinded (with paint on the eyes). However, the use of a rubber mat to reduce vibrations significantly decreased predation success rate. In addition, the presence or absence of the blindfold did not affect the predation rates under the reducing vibration condition. These results indicate that substrate vibrations emitted from prey are critically important to trigger the predatory behavior in L. typica, but visual and chemical stimuli are not used even in the case when vibration cues are unavailable. This is the first study to use vibration‐reducing experiments to experimentally demonstrate the critical cues for predation in trapdoor spiders.

Paullinia cupana (Kunth) stimulates behavior patterns and regulates oxidative stress markers in lobster cockroach Nauphoeta cinerea

Paullinia cupana (Kunth), popularly known as guarana, is a plant species from the Amazon Region of Brazil that stands out for being one of the most promising herbal medicines of the Brazilian flora since it has relevant biological properties. However, studies are still needed to use this species as a direct approach to evaluate aspects related to behavior and oxidative stress in invertebrate model animals. In this context, we aimed to evaluate the stimulant and the antioxidant effects of P. cupana in lobster cockroach Nauphoeta cinerea. For that, cockroaches were exposed to a diet separately supplemented by the decoction of P. cupana powder decoction at increasing concentrations of 1, 25, 50 and 100 mg/g for 42 days. Behavioral and biochemical assays were performed, respectively, to assess the locomotor/exploratory performance and oxidative stress marker levels of the cockroaches. They exhibited an outstanding increase in the locomotion performance and in the cell viability content, as well as in the protein and non-protein thiol levels. Moreover, there was a decrease in lipid peroxidation levels and in free Fe2+ ion contents. Together, our results demonstrate the stimulant and the antioxidant capacity of P. cupana by acting positively in behavioral patterns and by regulating oxidative stress markers in lobster cockroach N cinerea. These findings encourage further laboratory analyzes in order to better enlighten the specific mechanisms of action attributed to guarana.

Ovarian apoptosis is regulated by carbohydrate intake but not by protein intake in speckled cockroaches

When the likelihood of reproducing successfully is low, any prior investment in developing oocytes may be wasted. One means of recouping this investment is oosorption – where ova are absorbed and resources salvaged so they can be re-allocated to other traits. Food-limited female speckled cockroaches (Nauphoeta cinerea) appear to use this strategy. However, it is unclear if total food intake or the availability of specific nutrients induces this process. Here, we used the geometric framework of nutrition to determine how protein, carbohydrate and energy intake affect levels of ovarian apoptosis and necrosis (controlled versus uncontrolled cell death) in the terminal oocytes of female N. cinerea. We then compare the effects of nutrient intake on apoptosis (a key step towards oosorption) and offspring production to better understand the relationship between diet, apoptosis and female fitness. We found that even when food was abundant, females experienced high levels of apoptosis if their diet lacked carbohydrate. Necrosis was reduced when energy intake was high, but largely irrespective of nutrient ratio. Offspring production peaked on a low protein, high carbohydrate nutrient ratio (1P:7.96C), similar to that which minimized apoptosis (1P:7.34C) but not in the region of nutrient space that minimized necrosis. Thus, females consuming an ideal nutrient blend for reproduction can invest heavily in their current brood without needing to salvage nutrients from developing ova. However, offspring production was more dependent on carbohydrate consumption than apoptosis was, suggesting that the importance of carbohydrate in reproduction goes beyond regulating oosorption. This reliance on carbohydrate for female reproduction may reflect the unusual reproductive and nutritional physiology of speckled cockroaches; attributes that make this species an exciting model for understanding how diet regulates reproduction.

Identification and Semi-quantification of Protozoa from the Digestive System Microbiota of the Lobster Cockroach Nauphoeta cinerea Oliver, 1789 (Insecta:Blattaria).

The lobster cockroach Nauphoeta cinerea (N. cinerea) is indicated as a promising non-mammalian model, because it presents behavioral and biochemical alterations also observed in conventional models. In this research, we identified and characterized the distribution of protozoa that inhabit the digestive system (DS) of N. cinerea cockroaches. The adult specimens of N. cinerea used in this study (n = 32) were obtained at the Federal University of Santa Maria, dissected and had their visceral contents observed in bright-field microscopy without staining and after application of lugol, Ziehl-Neelsen staining, EA36 trichrome and simulated dark-field microscopy with application of nankin ink. The presence of protozoa in different portions of the DS was semi-quantified by a system of crosses (+). The main taxa observed were: amoebas (Archaemebae:Entamoebida), gregarins (Apicomplexa:Eugregarinide), coccidia (Apicomplexa:Eucoccidiorida), kinetoplastids (Kinetoplastea:Kinetoplastida) and oxymonads (Preaxostyla:Oxymonadida). The highest prevalence of amoebas and gregarines was observed in the medial portion of the DS, while for the other groups, this was seen in the final portion, and in the case of coccidia, such prevalence was specially evidenced by the alcohol-acid coloration. In the present work, the great biological diversity that exists in the microbiota of the digestive system of Nauphoeta cinerea was demonstrated, being possible to find several pathogenic species for humans such as Entamoeba histolytica/dispar/moshkovskii, Cryptosporidium sp. and Cyclospora cayetanensis. There is still a lot to know about the interactions between endocommensal protozoa and their respective invertebrate hosts, so the best way to clarify such relationships is through molecular and genetic test.

Identification and Validation of Reference Genes for RT-qPCR Normalization in Nauphoeta cinerea (Olivier, 1789) (Blattodea, Blaberidae)

Quantitative RT-PCR (q-RT-PCR) is a powerful tool that allows large-scale analysis of very small changes in gene expression. For the calculation of gene expression, such as the delta-delta Ct method, different PCR primer efficiencies (E) may affect the result, as PCR primer yields are assumed to be comparable for the gene of interest and housekeeping gene. Therefore, identification of a suitable reference gene for data normalization is an important step in the development of qPCR assays. Furthermore, accurate and reliable results depend on the use of stable reference genes for normalization. The aim of the current study is the identification and validation of a set of six housekeeping genes (GADPH, RPS18, α-TUB, EF1α, ArgK, and ACTB) in cockroach species Nauphoeta cinerea adults using five different algorithms (ΔCt method, Bestkeeper, geNorm, Normfinder and RefFinder) to evaluate the stability of selected reference genes expression. Our results show that α-Tub use provides accurate normalization of gene expression levels in N. cinerea adults. In addition, since the GADPH is selected as the second most stable reference gene, GADPH can be also used for transcript analysis N. cinerea adults. Our study also showed that ACTB (β-actin) should not be used for normalizing transcript levels when examining N. cinerea adults. Additionally, validation studies for reference genes in cockroaches are very few (only one) in the literature. Therefore, the results highlight the need for validation of reference genes under biotic and abiotic conditions in q-RT-PCR studies in cockroaches.

Genome size evaluations in cockroaches: new entries.

In this paper, we report genome size (GS) values for nine cockroaches (order Blattodea, families Blattidae, Blaberidae and Ectobiidae, ex Blattelidae), three of which are original additions to the ten already present in the GS database: the death’s head roach (Blaberus craniifer), the Surinam cockroach (Pycnoscelus surinamensis) and the Madeira cockroach (Leucophaea maderae). Regarding the American cockroach (Periplaneta americana), the GS database contains two contrasting values (2.72 vs 3.41 pg); likely, the 2.72 pg value is the correct one as it is strikingly similar to our sperm DNA content evaluation (2.80 ± 0.11 pg). Also, we suggest halving the published GS of the Argentine cockroach Blaptica dubia and the spotted cockroach (the gray cockroach) Nauphoeta cinerea discussing i) the occurrence of a correlation between increasing 2n chromosome number and GS within the order Blattodea; and ii) the possible occurrence of a polyploidization phenomenon doubling a basic GS of 0.58 pg of some termite families (superfamily Blattoidea, epifamily Termitoidae).

Neurolocomotor Behavior and Oxidative Stress Markers of Thiazole and Thiazolidinedione Derivatives against Nauphoeta cinerea.

Thiazolidine compounds NJ20 {(E)-2-(2-(5-bromo-2-methoxybenzylidene)hydrazinyl)-4-(4-nitrophenyl)thiazole} and NW05 [(2-(benzo (d) (1,3) dioxol-4-ylmethylene)-N-(4-bromophenyl)-thiosemicarbazone] potentiated the effect of norfloxacin in resistant bacteria; however, there are no reports on their effects on Nauphoeta cinerea in the literature. The objective of this work was to evaluate the behavioral effects and oxidative markers of NW05 and NJ20 in lobster cockroach N. cinerea. To evaluate the behavioral study, a video tracking software was used to evaluate the locomotor points and the exploratory profile of cockroaches in the horizontal and vertical regions of a new environment. The total concentration of thiol and reduced glutathione (GSH), substances reactive to thiobarbituric acid (TBARS), free iron (II) content and mitochondrial viability were determined. The antioxidant potential was evaluated by the DPPH method. Both substances induced changes in the behavior of cockroaches, showing a significant reduction in the total distance covered and in the speed. In the cell viability test (MTT), there was a significant reduction for NJ20 (1 mM). NJ20 caused a significant increase in total levels of thiol and non-protein thiol (NPSH), although it also slightly increased the content of malondialdehyde (MDA). Both compounds (NW05 and NJ20) caused a significant reduction in the content of free iron at a concentration of 10 mM. In conclusion, the compound NJ20 caused moderate neurotoxicity (1 mM), but had good antioxidant action, while NW05 did not show toxicity or significant antioxidant activity in the model organism tested. It is desirable to carry out complementary tests related to the antioxidant prospection of these same compounds, evaluating them at different concentrations.

Cockroach (Blattodea, Blaberoidea) fauna of the Maltese Islands, with descriptions of two new species

Cockroach (Blattodea, Blaberoidea) fauna of the Maltese Islands. Hitherto seven species of cockroaches were known from the Maltese Archipelago (Schembri 1980): four of them are synanthropic species introduced by man: Blatta orientalis, Blattella germanica, Periplaneta americana and Supella longipalpa; the remaining three are indigenous species: Ectobius kraussianus, Loboptera decipiens and Polyphaga aegyptiaca. The earlier reports almost exclusively concern the largest island Malta; only one report (L. decipiens) applies to the islet Comino. Faunistic part. By recent collectings of the authors (mainly A.S.) including also the smaller islands Gozo, Comino and some rock islets (Cominotto, Filfla, Fungus Rock, Large Blue Lagoon Rock, Pigean Rock) the known distribution of the species could be largely widened; two further obviously indigenous species were found, both new to science and presumably endemic to Malta: Ectobius melitensis, sp. nov. (Ectobiidae, Ectobiinae) and Heterogamisca jeffreyana, sp. nov. (Corydiidae, Corydiinae). E. kraussianus presumably has to be eliminated from the list of Maltese cockroaches; the older report most likely concerns a misidentified E. melitensis. During the last years the number of introduced species was strongly increased (1) by escapees of species cultivated for the nourishment of amphibia and reptilia, as are Blaptica dubia, Gromphadorhina sp., Nauphoeta cinerea, Periplaneta fuliginosa, Shelfordella lateralis and Symploce pallens, and (2) by species presumably imported with gardening materials: Pycnoscelus surinamensis and Phoetalia pallida. The mentioned species have repeatedly been found in human buildings and adjacent gardens of a number of settlements, mostly in the surroundings of the capital Valetta. They seem to be well established there; part of them has even been observed in the wild. The newly introduced species are in spite of their successful establishment and reproduction in Malta not considered as a danger for the indigenous species: The great differences in the requirements between the two groups most likely exclude competition between them. Three species, Ectobius vittiventris, Henschoutedenis flexivitta, and Periplaneta australasiae, have only occasionally been found and can, therefore, scarcely be considered as members of the Maltese fauna; the latter also applies to species of the genus Panchlora which were repeatedly observed on fruit markets at freshly imported bananas, but never found outside, neither in the Maltese banana plantations. Taxonomic part. The new species are described and compared with their closest relatives, colouration and characteristic structures are shown in several figures. Ectobius melitensis belongs to the kraussianus-species group of Ectobius known from Sicily and surrounding islands (Ustica, Aeolian Islands, Ponza), but with one species also reaching Albania; the nearest relatives of Heterogamisca jeffreyana are occurring on the Iberian Peninsula and North Africa (Morocco, Algeria, Tunisia).

Streptozotocin activates inflammation-associated signalling and antioxidant response in the lobster cockroach; Nauphoeta cinerea (Blattodea: Blaberidae)

Streptozotocin exhibits tropism to insulin-producing beta-cells in mammals and has been used to model diabetes-like phenotypes in insects. We have previously shown increased brain glucose levels and oxidative stress in STZ-treated nymphs of Nauphoeta cinerea. Here, we validate Nauphoeta cinerea as an experimental organism for studying STZ-induced metabolic disruptions by investigating the potential changes in the expression of inflammation and antioxidant related genes. Cockroaches were injected with 0.8% NaCl, 74 and 740 nmol of STZ. mRNA extracted from the head of cockroaches was used to estimate the RT-qPCR expression of inflammation and antioxidant genes. STZ-treatment upregulated the target genes of the JNK pathway (early growth factor response factor and reaper) but had no effect on PDGF-and VEGF-related factor 1. TOLL 1, the target gene of TOLL/NF-kB pathway was up regulated, while both the activator and target gene of the UPD3/JAK/STAT pathway [unpaired 3 and Suppressor of cytokine signalling at 36E] were upregulated. mRNA levels of primary antioxidants (superoxide dismutase and catalase) were increased in STZ treated nymphs but there was no effect on thioredoxins and Peroxiredoxin 4. Likewise, STZ treatment did not affect the expression of the delta class of the glutathione S-transferase gene family, but the sigma and theta classes of the GST family were upregulated. The STZ-induced N. cinerea gene expression modification demonstrates the involvement of primary antioxidants and the GST detoxification system in the cockroach oxidative stress response and buttresses the proposed crosstalk between inflammatory and redox pathways.

Social interactions upregulate hemolymph tryptophan and tyrosine levels in the male lobster cockroach

In the present study, we found that tryptophan (TRP) and tyrosine (TYR) levels are increased in hemolymph of male Nauphoeta cinerea after social contact with either male or female conspecifics. Hemolymph was collected from individual males before and after the social interactions, and samples were analyzed by HPLC-ECD; analyte identities were confirmed by UPLC/MS. After a male-male first encounter fight, hemolymph TRP and TYR levels were significantly increased in dominants compared with the levels before the encounter. Conversely, TRP and TYR in subordinates were maintained at levels similar to those before the encounter. While after-fight TRP and TYR levels were significantly higher in dominants than subordinates, no significant differences were found in the contestants before the fight. Moreover, contact with an isolated male antenna was sufficient to stimulate attack behavior and increase hemolymph TRP and TYR titers to levels similar to those seen in dominants. After a male-female interaction, two distinct outcomes could be observed. Either hemolymph TRP and TYR levels were increased in successfully mated males, or TRP and TYR levels were unchanged in males that only exhibited premating wing-raising behavior but failed in mating. After contacting the antenna of a socially naïve male with an isolated female antenna, three patterns of behavior and related amino acid response were observed: 1) only premating wing-raising behavior with significant increase of TRP and TYR levels, 2) only attack behavior with significant increase of TRP and TYR levels, and 3) mixed wing-raising and attack behaviors with no significant changes in TRP and TYR levels. The present results show a robust response of hemolymph TRP and TYR to social contact. In light of previously characterized responses in pheromone and juvenile hormone levels, these amine responses suggest that the physiological response of N. cinerea to social contact is multi-dimensional.

Chronic ciprofloxacin and atrazine co-exposure aggravates locomotor and exploratory deficits in non-target detritivore speckled cockroach (Nauphoeta cinerea).

The global detection of ciprofloxacin and atrazine in soil is linked to intensive anthropogenic activities in agriculture and inadvertent discharge of industrial wastes to the environment. Nauphoeta cinerea is a terrestrial insect with cosmopolitan distribution and great environmental function. The current study probed the neurobehavioral and cellular responses of N. cinerea singly and jointly exposed to atrazine (1.0 and 0.5μgg-1 feed) and ciprofloxacin (0.5 and 0.25μgg-1 feed) for 63days. Results demonstrated that the reductions in the body rotation, maximum speed, turn angle, path efficiency, distance traveled, episodes, and time of mobility induced by atrazine or ciprofloxacin per se was exacerbated in the co-exposure group. The altered exploratory and locomotor in insects singly and jointly exposed to ciprofloxacin and atrazine were verified by track plots and heat maps. Furthermore, we observed a decrease in acetylcholinesterase and anti-oxidative enzyme activities with concomitant elevation in the levels of lipid peroxidation, nitric oxide, and reactive oxygen and nitrogen species were significantly intensified in the midgut, hemolymph, and head of insects co-exposed to ciprofloxacin and atrazine. In conclusion, exposure to binary mixtures of ciprofloxacin and atrazine elicited greater locomotor and exploratory deficits than upon exposure to the individual compound by inhibiting acetylcholinesterase activity and induction of oxido-inflammatory stress responses in the insects. N. cinerea may be a usable model insect for checking contaminants of ecological risks.

PSVI-26 Insect meals as novel protein sources in wet pet foods for adult cats

Abstract As the human and pet populations increase, the human and pet food industries will experience a greater demand for animal-based protein sources. Finding alternative proteins that are nutritional adequate and safe for companion animals may contribute to diversification of protein sources and lower the demand for animal-based protein in pet food products. However, limited information is available on the nutritional value of insects for pet animals. Thus, the object of this study was to evaluate three different insect meals, Speckled cockroach (SC; Nauphoeta cinerea), Madagascar hissing cockroach (MC; Gromphadorhina portentosa) and superworm (SW; Zophobas morio larvae), added at the expense of chicken meal (control diet), as protein sources in retorted feline diets. This study was a complete randomized design, with a total of 28 cats (mean age = 2.1 ± 0.03 yr; mean BW= 4.9 ± 0.8 kg) randomly assigned to 1 of the 4 dietary treatments. All animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee. All diet formulated meet or exceed the AAFCO (2018) nutrient profile. The experimental period was 28 d in length, with 7 d of diet adaptation (control diet) followed by 21 of feeding the assigned experimental diets. Fecal samples were collected at the last 4 d of experiment. All diets were well digested by the cats, and apparent total tract digestibility of dry matter (86.5–88.1%), organic matter (88.9–90.6%), fat (90.1–92.3%), crude protein (86.3–89.4%) did not differ among treatments (P > 0.05). Fecal scores were not affected by dietary treatment, ranging from 1.8 to 2.2 (5–point scale). Similarly, fecal branched-chain fatty acids, indole and phenol concentrations did not differ among treatments. Overall, the selected insect meals tested herein had no negative effects on macronutrient digestibility, fecal characteristics and metabolites, or overall health of adult cats.

Streptozotocin induces brain glucose metabolic changes and alters glucose transporter expression in the Lobster cockroach; Nauphoeta cinerea (Blattodea: Blaberidae).

The development of new models to study diabetes in invertebrates is important to ensure adherence to the 3R's principle and to expedite knowledge of the complex molecular events underlying glucose toxicity. Streptozotocin (STZ)-an alkylating and highly toxic agent that has tropism to mammalian beta cells-is used as a model of type 1 diabetes in rodents, but little is known about STZ effects in insects. Here, the cockroach; Nauphoeta cinerea was used to determine the acute toxicity of 74 and 740nmol of STZ injection per cockroach. STZ increased the glucose content, mRNA expression of glucose transporter 1 (GLUT1) and markers of oxidative stress in the head. Fat body glycogen, insect survival, acetylcholinesterase activity, triglyceride content and viable cells in head homogenate were reduced, which may indicate a disruption in glucose utilization by the head and fat body of insects after injection of 74 and 740nmol STZ per nymph. The glutathione S-transferase (GST) activity and reduced glutathione levels (GSH) were increased, possibly via activation of nuclear factor erythroid 2 related factor as a compensatory response against the increase in reactive oxygen species. Our data present the potential for metabolic disruption in N. cinerea by glucose analogues and opens paths for the study of brain energy metabolism in insects. We further phylogenetically demonstrated conservation between N. cinerea glucose transporter 1 and the GLUT of other insects in the Neoptera infra-class.

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.

Toxicological outcome of exposure to psychoactive drugs carbamazepine and diazepam on non-target insect Nauphoeta cinerea

The continuous detection of human pharmaceuticals during environmental biomonitoring is a global concern because of the menaces they may exert on non-target organisms. Carbamazepine (CBZ) and diazepam (DZP) are commonly prescribed psychotropic drugs which have been reported to coexist in the environment globally. Nauphoeta cinerea is a common insect with high ecological impact. This study elucidated the influence of co-exposure to DZP (0.5 and 1.0 μg kg−1 diet) and CBZ (1.5 and 3.0 μg kg−1 diet) for 42 days on the behavior and biochemical responses in Nauphoeta cinerea. Results showed that DZP alone did not induce adverse effect on the behavior and antioxidant status in the exposed insects. However, exposure to CBZ alone and binary mixtures of DZP and CBZ significantly decreased locomotor and exploratory accomplishments evidenced by decreased mobile episodes, total mobile time, maximum speed, total distance traveled, absolute turn angle, body rotation and path efficiency in comparison with control. The decline observed in the exploratory activities of insects fed with CBZ alone and the mixtures was confirmed by track plots and heat maps. Further, acetylcholinesterase and antioxidant enzyme activities decreased significantly whereas reactive oxygen and nitrogen species, nitric oxide and lipid peroxidation levels increased significantly in the hemolymph, head and midgut of insects exposed to CBZ alone and the mixtures. Collectively, CBZ alone and binary mixtures of CBZ and DZP caused neurotoxicity via induction of inflammatory and oxidative stress in insects. Nauphoeta cinerea may be a potential non-target insect model for monitoring ecotoxicological hazard of pharmaceuticals.

Water Costs of Gas Exchange by a Speckled Cockroach and a Darkling Beetle.

Respiratory water loss during metabolic gas exchange is an unavoidable cost of living for terrestrial insects. It has been suggested to depend on several factors, such as the mode of gas exchange (convective vs. diffusive), species habitat (aridity), body size and measurement conditions (temperature). We measured this cost in terms of respiratory water loss relative to metabolic rate (respiratory water cost of gas exchange; RWL/V˙CO2) for adults of two insect species, the speckled cockroach (Nauphoeta cinerea) and the darkling beetle (Zophobas morio), which are similar in their mode of gas exchange (dominantly convective), habitat (mesic), body size and measurement conditions, by measuring gas exchange patterns using flow-through respirometry. The speckled cockroaches showed both continuous and discontinuous gas exchange patterns, which had significantly a different metabolic rate and respiratory water loss but the same respiratory water cost of gas exchange. The darkling beetles showed continuous gas exchange pattern only, and their metabolic rate, respiratory water loss and respiratory cost of gas exchange were equivalent to those cockroaches using continuous gas exchange. This outcome from our study highlights that the respiratory water cost of gas exchange is similar between species, regardless of gas exchange pattern used, when the confounding factors affecting this cost are controlled. However, the total evaporative water cost of gas exchange is much higher than the respiratory cost because cuticular water loss contributes considerably more to the overall evaporative water loss than respiratory water. We suggest that the total water cost of gas exchange is likely to be a more useful index of environmental adaptation (e.g., aridity) than just the respiratory water cost.

Cockroaches: an alternative model to teach enzymatic inhibition to undergraduate students

ABSTRACT Enzymes are organic molecules known for their catalytic nature because they accelerate metabolic reactions inorganisms. The study of enzyme functions and modes of action is essential to understand Biochemistry. This study aimed to analyse the acetylcholinesterase enzymatic activity, as well as the enzymatic inhibition mechanism in practical classes. The enzyme was obtained from the cockroach (Nauphoeta cinerea) head, an alternative model to the use of vertebrates in practical classes. For this study, practical classes were carried out in two undergraduate courses: Biology and Veterinary Medicine. It was observed the knowledge consolidation through the analysis of the answers given to the questionnaire. The students’ enthusiasm to work with animals was evident. This fact led to a better understanding about the subject and more attractive classes.

Dietary monosodium glutamate altered redox status and dopamine metabolism in lobster cockroach (Nauphoeta cinerea).

Monosodium Glutamate (MSG) is the most commonly utilized food additive in the world. However, data on possible biochemical reasons underlying the neurotoxic effects of dietary MSG is limited. Therefore, this study investigated the effects of dietary supplementation of MSG on redox status and neurochemical indices in lobster cockroach nymph. These were evaluated via assessment of enzymatic and nonenzymatic antioxidants, acetylcholinesterase and monoamine oxidase activities, and dopamine content in the cockroach nymph head homogenate. MSG supplemented diet caused dose-dependent significant (p<.05) reduction in % survival, thiol, GSH, dopamine contents, and GST activity, increased ROS, NO, Fe2+ , MDA contents, and MAO activity but no significant (p<.05) difference was obtained in GSH and TBARS contents, and AChE activity. Increased oxidative, cholinergic, and monoaminergic activities coupled with decreased dopamine level might be the plausible biochemical explanation for the neurotoxic effects observed during sub-chronic consumption of large amounts of MSG in diet. PRACTICAL APPLICATIONS: This study suggests that consumption of monosodium glutamate should be reduced to the barest minimum due to its capability to induce oxidative stress and nervous toxicological effects at high dosage.