Oxidative stress induction by essential oil from Piper alatipetiolatum (Piperaceae) triggers lethality in the larvae of Culex quinquefasciatus (Diptera: Culicidae).

Culex quinquefasciatus is the main vector of lymphatic filariasis in Brazil, which present resistance to commercial insecticides. Nowadays, essential oils (EOs) exhibiting larvicidal activity, such as those derived from Piper alatipetiolatum, provide a promising alternative for vector control, including Culex species. This study aimed to investigate the larvicidal activity and the oxidative stress indicators of the EO from P. alatipetiolatum in Cx. quinquefasciatus larvae. The EO was extracted from P. alatipetiolatum leaves using the hydrodistillation method, resulting in a yield of 7.2 ± 0.1%, analysed by gas chromatography coupled with spectrometry and gas chromatography coupled with flame ionization detector (GC-MS and GC-FID), and evaluated against Cx. quinquefasciatus larvae. Reactive Oxygen and Nitrogen Species (RONS), Catalase (CAT), glutathione-S-transferase (GST), acetylcholinesterase (AChE), and Thiol levels were used as oxidative stress indicators. Analysis by CG-MS and CG-FID revealed that the main compound in the EO was the oxygenated sesquiterpene ishwarone, constituting 78.6% of the composition. Furthermore, the EO exhibited larvicidal activity, ranging from 26 to 100%, with an LC50 of 4.53 µg/mL and LC90 of 15.37 µg/mL. This activity was accompanied by a significant increase in RONS production, alterations in CAT, GST, AChE activity, and thiol levels compared to the control groups (p < 0.05). To the best of our knowledge, this is the first report describing the larvicidal activity and oxidative stress induced by the EO from P. alatipetiolatum against Cx. quinquefasciatus larvae. Therefore, we propose that this EO shows promise as larvicidal agent for the effective control of Cx. quinquefasciatus larvae.

Essential oil and fenchone extracted from Tetradenia riparia (Hochstetter.) Codd (Lamiaceae) induce oxidative stress in Culex quinquefasciatus larvae (Diptera: Culicidae) without causing lethal effects on non-target animals.

We investigated the larvicidal activity of the essential oil (EO) from Tetradenia riparia and its majority compound fenchone for controlling Culex quinquefasciatus larvae, focusing on reactive oxygen and nitrogen species (RONS), catalase (CAT), glutathione S-transferase (GST), acetylcholinesterase (AChE) activities, and total thiol content as oxidative stress indicators. Moreover, the lethal effect of EO and fenchone was evaluated against Anisops bouvieri, Diplonychus indicus, Danio rerio, and Paracheirodon axelrodi. The EO and fenchone (5 to 25 µg/mL) showed larvicidal activity (LC50 from 16.05 to 18.94 µg/mL), followed by an overproduction of RONS, and changes in the activity of CAT, GST, AChE, and total thiol content. The Kaplan-Meier followed by Log-rank (Mantel-Cox) analyses showed a 100% survival rate for A. bouvieri, D. indicus, D. rerio, and P. axelrodi when exposed to EO and fenchone (262.6 and 302.60 µg/mL), while α-cypermethrin (0.25 µg/mL) was extremely toxic to these non-target animals, causing 100% of death. These findings emphasize that the EO from T. riparia and fenchone serve as suitable larvicides for controlling C. quinquefasciatus larvae, without imposing lethal effects on the non-target animals investigated.

Essential oil from Piper tuberculatum Jacq. (Piperaceae) and its majority compound ß-caryophyllene: mechanism of larvicidal action against Aedes aegypti (Diptera: Culicidae) and selective toxicity.

Synthetic insecticides have been the primary approach in controlling Aedes aegypti; however, their indiscriminate use has led to the development of resistance and toxicity to non-target animals. In contrast, essential oils (EOs) are alternatives for vector control. This study investigated the mechanism of larvicidal action of the EO and ß-caryophyllene from Piper tuberculatum against A. aegypti larvae, as well as evaluated the toxicity of both on non-target animals. The EO extracted from P. tuberculatum leaves was majority constituted of ß-caryophyllene (54.8%). Both demonstrated larvicidal activity (LC50 of 48.61 and 57.20 ppm, p < 0.05), acetylcholinesterase inhibition (IC50 of 57.78 and 71.97 ppm), and an increase in the production of reactive oxygen and nitrogen species in larvae after exposure to the EO and ß-caryophyllene. Furthermore, EO and ß-caryophyllene demonstrate no toxicity to non-target animals Toxorhynchites haemorrhoidalis, Anisops bouvieri, and Diplonychus indicus (100% of survival rate), while the insecticide α-cypermethrin was highly toxic (100% of death). The results demonstrate that the EO from P. tuberculatum and ß-caryophyllene are important larvicidal agents.

Toxicity of the essential oil from Tetradenia riparia (Hochstetter.) Codd (Lamiaceae) and its principal constituent against malaria and dengue vectors and non-target animals.

Malaria and dengue are diseases transmitted by mosquitoes of the genera Anopheles and Aedes resistant to commercial insecticides, which are toxic to non-target animals. Alternatively, eco-friendly strategies have focused on searching for essential oil (EO) from plants to control these mosquitoes. In this aspect, this study was carried out to investigate the toxicity of the EO from Tetradenia riparia and its main constituent against Anopheles and Aedes larvae and non-target animals Toxorhynchites haemorrhoidalis and Gambusia affinis. The mechanism of the larvicidal action of the EO and its main compound was investigated by the acetylcholinesterase (AChE) inhibition. The EO from T. riparia was extracted by hydrodistillation with yield of 1.4 ± 0.17%. The analysis of the EO by GC-MS and GC-FID revealed fenchone (38.62%) as the main compound. The EO (100 ppm) showed larvicidal activity against Anopheles and Aedes larvae (91 to 100% of mortality) (LC50 from 29.31 to 40.76 ppm). On the other hand, fenchone (10 ppm) showed more activity (89 to 100% of mortality) (LC50 from 5.93 to 7.00 ppm) than the EO. The EO and fenchone caused the inhibition of AChE (IC50 from 1.93 to 2.65 ppm), suggesting the inhibition of this enzyme as a possible mechanism of larvicidal action. Regarding toxicity, the EO (1000 ppm) and fenchone (100 ppm) showed low toxicity against T. haemorrhoidalis and G. affinis (9 to 74% of mortality) (LC50 from 170.50 to 924.89 ppm) (SI/PSF from 17.99 to 31.91) than the α-cypermethrin (0.52 ppm) which was extremally toxic against these non-target animals (100% of mortality, LC50 from 0.22 to 0.29 ppm). This significant larvicidal activity of the T. riparia EO and its main constituent, along with the low toxicity towards non-target organisms indicate these samples as a possible eco-friendly alternative for the control of malaria and dengue vectors.

Essential oil of Piper purusanum C.DC (Piperaceae) and its main sesquiterpenes: biodefensives against malaria and dengue vectors, without lethal effect on non-target aquatic fauna.

The mosquito vectors of the genera Aedes and Anopheles present resistance to several commercial insecticides, which are also toxic to non-predator targets. On the other hand, essential oils are a promising source of insecticides. Thus, in this work, the essential oil from the leaves of Piper purusanum was characterized by gas chromatography-based approaches and evaluated as biodefensive against malaria and dengue vectors. The main compounds of P. purusanum essential oil were ß-caryophyllene (57.05%), α-humulene (14.50%), and germacrene D (8.20%). The essential oil inhibited egg hatching (7.6 ± 1.5 to 95.6 ± 4.5%), caused larval death (LC50 from 49.84 to 51.60 ppm), and inhibited the action of acetylcholinesterase (IC50 of 2.29 µg/mL), which can be related to the mechanisms of action. On the other hand, the biological activities of ß-caryophyllene, α-humulene, and germacrene D were higher than that of essential oil. In addition, these sesquiterpenes and essential oil did not show a lethal effect on Toxorhynchites splendens, Anisops bouvieri, Gambusia affinis, and Diplonychus indicus (LC50 from 2098.80 to 7707.13 ppm), although D. indicus is more sensitive (SI/PSF from 48.56 to 252.02 ppm) to essential oil, representing a natural alternative against these relevant vectors.

Production of thermostable ß-glucosidase and CMCase by Penicillium sp: LMI01 isolated from the Amazon region

Background: Cellulolytic enzymes of microbial origin have great industrial importance because of their wide application in various industrial sectors. Fungi are considered the most efficient producers of these enzymes. Bioprospecting survey to identify fungal sources of biomass-hydrolyzing enzymes from a high-diversity environment is an important approach to discover interesting strains for bioprocess uses. In this study, we evaluated the production of endoglucanase (CMCase) and ß-glucosidase, enzymes from the lignocellulolytic complex, produced by a native fungus. Penicillium sp. LMI01 was isolated from decaying plant material in the Amazon region, and its performance was compared with that of the standard isolate Trichoderma reesei QM9414 under submerged fermentation conditions. Results: The effectiveness of LMI01 was similar to that of QM9414 in volumetric enzyme activity (U/mL); however, the specific enzyme activity (U/mg) of the former was higher, corresponding to 24.170 U/mg of CMCase and 1.345 U/mg of ß-glucosidase. The enzymes produced by LMI01 had the following physicochemical properties: CMCase activity was optimal at pH 4.2 and the ß-glucosidase activity was optimal at pH 6.0. Both CMCase and ß-glucosidase had an optimum temperature at 60°C and were thermostable between 50 and 60°C. The electrophoretic profile of the proteins secreted by LMI01 indicated that this isolate produced at least two enzymes with CMCase activity, with approximate molecular masses of 50 and 35 kDa, and ß-glucosidases with molecular masses between 70 and 100 kDa. Conclusions: The effectiveness and characteristics of these enzymes indicate that LMI01 can be an alternative for the hydrolysis of lignocellulosic materials and should be tested in commercial formulations.

Occurrence of Aelurostrongylus abstrusus (Railliet, 1898) larvae (Nematoda: Metastrongylidae) infecting Achatina (Lissachatina) fulica Bowdich, 1822 (Mollusca: Gastropoda) in the Amazon region / Ocorrência de Aelurostrongylus abstrusus (Railliet, 1898) (Nematoda: Metastrongylidae) infectando o Achatina (Lissachatina) fulica Bowdich, 1822 (Mollusca: Gastropoda) na região amazônica

Achatina fulica or "giant African snail" is an exotic species, considered to be one of the world's hundred most invasive species, causing serious environmental damages. In the present study we report, for the first time, the occurrence of Aelurostrongylus abstrusus infecting A.fulica in the Amazon region. This nematode is described parasitizing mainly the pulmonary system of felines, which causes "aelurostrongilose", also known as feline cardio-pulmonary strongyloidosis. New morphometric data of third stage larvae are presented herein. The present study demonstrated that 40 percent of all the snails were infected by A. abstrusus. Achatina fulica specimens were collected from three different areas in Manaus namely: rural; east and west areas. The east area presents the highest prevalence of 80 percent. The large number of A.fulica found in inhabited areas increases the chances of emergent zoonoses, which highlights the need of further studies so as to better control this disease.

Achatina fulica ou "caramujo africano" é uma espécie exótica, considerada uma das cem piores espécies invasoras do mundo, causando sérios danos ambientais. No presente estudo foi registrado, pela primeira vez, a ocorrência do Aelurostrongylus abstrusus infectando o A.fulica na região amazônica. Esse nematóide é descrito parasitando principalmente o sistema pulmonar de felinos, causando a "aelurostrongilose", também conhecida como estrongiloidose cardio-pulmonar felina. Novos dados morfométricos de larvas de terceiro estágio são apresentados. Dos 45 caramujos coletados, 40 por cento estavam infectados por larvas de A. abstrusus. Especimens de Achatina fulica foram coletados em três áreas da cidade de Manaus: rural, leste e oeste. A zona leste apresentou a maior prevalência de 80 por cento. O grande número de A.fulica encontrado em áreas habitadas aumenta as chances de ocorrência de zoonoses emergentes e destaca a necessidade de mais estudos para o melhor controle da doença.

Calibration and evaluation of field cage for oviposition study with Aedes (Stegomyia) aegypti female (L.) (Diptera: Culicidae) / Calibração e avaliação de semi-campo para estudos de oviposição para fêmeas de Aedes (Stegomyia) aegypti (L.) (Diptera: Culicidae)

Differences among results gathered from insect behavior studies conducted in laboratory and field situations are due to ambient variables that differ greatly between both environments. In laboratory studies the environmental conditions can be controlled whereas in field temperature, humidity and air velocity vary uncontrollably. The objective of this study was to calibrate and evaluate an experimental area (field cage) (14 x 7 x 3.5 m) subdivided into eight test cages (2.5 x 2.5 x 2 m) for use in behavioral oviposition tests of Aedes aegypti (L.) mosquitoes for developing a new methodology to assess attractants and oviposition traps. Test cage calibration involved: (1) minimal experiment duration tests; (2) optimal female release number per traps test and (3) trap placement tests. All tests used gravid A. aegypti females; 3-4 days post blood meal and the sticky trap MosquiTRAP® to catch adults. Ninety percent of the females released were recaptured 2h after the beginning of the experiment, and this allowed up to 32 test repetitions/day to be conducted in the field cage. The minimum number of females necessary to conduct statistical analyses was 20 females/trap/test per cage. No significant difference was found in the behavioral response of gravid females to four different trap positions within test cages. Field trapping results with attractant were similar to those in the field cage. Therefore, the field cage could replace field trapping for evaluating at least mosquito traps and oviposition attractants for A. aegypti.

Discrepâncias entre resultados de estudos de comportamento de insetos em laboratório e em campo ocorrem devido a variáveis encontradas entre os ambientes. Enquanto em laboratório as condições são controladas, em campo os insetos são avaliados em condições variáveis de temperatura, umidade e velocidade do ar. O objetivo deste trabalho foi calibrar e avaliar uma área experimental (14 x 7 x 3,5 m) contendo oito gaiolas de teste (2,5 x 2,5 x 2 m), para estudos comportamentais de oviposição para Aedes aegypti (L.) visando desenvolver uma nova metodologia para avaliar atraentes e armadilhas de oviposição em semi-campo. A calibração consistiu na (1) determinação do tempo de realização dos experimentos e no (2) número de fêmeas liberadas por teste e na (3) avaliação das posições das armadilhas no interior das gaiolas de teste. Fêmeas de A. aegypti de 3-4 dias após repasto sangüíneo e MosquiTRAP® foram utilizadas nos experimentos. Noventa por cento das fêmeas liberadas foram recapturadas após 2h do início dos experimentos, permitindo a realização de até 32 repetições/dia. A amostra mínima de fêmeas por experimento para a realização de análises estatísticas foi limitada a 20 fêmeas/armadilha/gaiola. Não foi observada diferença na resposta comportamental das fêmeas nas diferentes posições avaliadas. Resultados com MosquiTRAP em área urbana foram semelhantes aos do semi-campo. Portanto, a metodologia pode substituir os testes de campo para avaliar pelo menos armadilhas e atrativos de oviposição para A. aegypti.