Encapsulation of carvacrol and thymol with yeast cell wall and its repellent activity against Amblyomma sculptum and Rhipicephalus sanguineus (Sensu Lato).

The main way to avoid contact with ticks and consequently tick-borne disease is the use of synthetic repellents. The search of new repellent compounds to increase the possibilities of use in strategies controls are necessary. The present study evaluated the repellent activity of two natural terpenes carvacrol and thymol in each one two different formulation (encapsulated and nonencapsulated with yeast cell wall) against the ticks Amblyomma sculptum and Rhipicephalus sanguineus sensu lato nymphs. Nymphs of A. sculptum and R. sanguineus s.l. of a single generation were used. The vertical filter paper repellency assay were performed with different concentration of both terpenes encapsulated and nonencapsulated in yeast cell wall. The repellent concentration 50% (RC50) were calculated to each compound formulation. Both carvacrol and thymol (encapsulated and nonencapsulated), had a repellent activity against A. sculptum and R. sanguineus s.l nymphs. Amblyomma sculptum was more sensitive to nonencapsulated carvacrol (RC50 values: 0.0032 to 0.0082 mg/cm2 after 1 and 15 min) (P < 0.05), while R. sanguineus s.l. was more sensitive to encapsulated carvacrol (RC50 values: 0.00008 to 0.0035 mg/cm2 after 1 and 15 min) (P < 0.05). Among tick species, R. sanguineus s.l. was more sensitive for most compounds than A. sculptum (P < 0.05). Although with distinct repellent activities, carvacrol and thymol encapsulated can be a promising alternative to synthetic repellents against A. sculptum and R. sanguineus s.l.

Repellent effects of Chinese cinnamon oil on nymphal ticks of Haemaphysalis longicornis, Rhipicephalus haemaphysaloides, and Hyalomma asiaticum.

The repellent activity of Chinese cinnamon oil (Cinnamomum cassia) on nymphal ticks (Haemaphysalis longicornis Neumann, Rhipicephalus haemaphysaloides Supino, and Hyalomma asiaticum Schulze and Schlottke) was evaluated in a sample Y-tube bioassay. The results were based on the vertical migration of ticks during the host-seek phase and showed a dose-dependent repellent effect of Chinese cinnamon oil on the tested nymphs after 6 h. For H. longicornis, R. haemaphysaloides, and H. asiaticum at the concentrations (vol/vol) of 3, 3, and 1.5%, the repellent percentages over time were 68-97, 69-94, and 69-93%, respectively, which indicated strong repellent activities against ticks, similar to the positive control DEET (N,N-diethyl-3-methylbenzamide). Chinese cinnamon oil exerted the strongest effect on H. asiaticum nymphs. To our knowledge, this is the first study to investigate the repellent effects of Chinese cinnamon oil on ticks. Chinese cinnamon oil has considerable potential and should be developed as a practical tick repellent.

Persistence Enhancement of a Promising Tick Repellent, Benzyl Isothiocyanate, by Yeast Microcarriers.

Phenethyl isothiocyanate isolated from Armoracia rusticana root oil and its derivatives were tested at different doses in a bioassay designed to evaluate repellency against individual Haemaphysalis longicornis nymphs. Among the tested compounds, benzyl isothiocyanate exhibited repellency against H. longicornis nymphs at the lowest dose of 0.00625 mg/cm2, followed by phenethyl isothiocyanate (0.0125 mg/cm2) and phenyl isothiocyanate (0.025 mg/cm2). The behavioral responses of H. longicornis nymphs exposed to benzyl isothiocyanate and phenethyl isothiocyanate indicated that the mode of action of these compounds can be mainly attributed to the vapor phase. Encapsulated benzyl isothiocyanate showed repellency up to 120 min post-application at 0.1 mg/cm2, whereas pure benzyl isothiocyanate showed repellency up to 60 min post-application at 0.1 mg/cm2. The present study suggests that benzyl isothiocyanate is a potential repellent for protection against H. longicornis nymphs, and encapsulation in yeast cells may enhance the repellency effect.

Ovicidal activity of spirotetramat and its effect on hatching, development and formation of Frankliniella occidentalis egg.

Frankliniella occidentalis (Pergande) has become an important vegetable pest worldwide because of its economic damage to crop production. However, it is difficult to control due to its unique living habits. In this study, the eggs of F. occidentalis were used as the target to explore the ovicidal activity of spirotetramat on the thrips and its effect on hatching, development and formation. After the treatment of spirotetramat, the LC50 value descreased with increased egg age using egg dipping method, and showed the same trend as the leaf dipping method verified on living plants. Through ultra-depth-of-field microscopy, scanning electron microscopy and transmission electron microscopy, the egg shell and internal structures of F. occidentalis eggs were studied. Spirotetramat can destroy the egg shells of F. occidentalis, resulting in shrinkage of the egg surface, sunken pores, egg deformities, egg shell rupture and other phenomena. This allows spirotetramat to enter the egg and destroy the egg structure, making the egg internal structure flocculent, fuzzy and unevenly distributed, which affects embryonic development and causes the nymphs to die before hatching. Therefore, the prevention and control of F. occidentalis using spirotetramat before damage is caused to crops should have a better effect.

Insecticidal efficacy of fluralaner (Bravecto®) against Triatoma brasiliensis, a major vector of Trypanosoma cruzi in Brazil.

BACKGROUND: Triatomines are responsible for the vector transmission of the protozoan parasite Trypanosoma cruzi, which causes Chagas disease. Triatoma brasiliensis is the main vector of the parasite in Brazil, and dogs are an important reservoir of the parasite. The aim of this study was to evaluate the insecticidal effect of fluralaner (Bravecto®) on T. brasiliensis after a blood meal in treated dogs. METHODS: Healthy mongrel dogs (n = 8) were recruited from the Zoonoses Control Center (ZCC) in the city of Natal, Rio Grande do Norte, Brazil, and randomized into two groups, a fluralaner (Bravecto®)-treated group (n = 4) and a control group (n = 4). Colony-reared third-, fourth- and fifth-instar nymphs of T. brasiliensis nymphs (n = 10) were allowed to feed on dogs from both groups for 30-40 min, once monthly, for up to 12 months. Bug mortality was observed up to 5 days after each blood meal. RESULTS: Mortality in triatomines which had a blood meal on fluralaner (Bravecto®)-treated dogs was 100% for up to 7 months after treatment, with mortality decreasing to 66.4% after 8 months, 57% after 9 months, 35% after 10 months, 10% after 11 months and 0% after 12 months. The mortality of triatomines that fed on non-treated control dogs was always ≤ 2.5%. CONCLUSIONS: Our results suggest that fluralaner (Bravecto®) treatment of dogs induces long-term mortality of T. brasiliensis after the blood meal. This is a potential approach to be used to control vector transmission of T. cruzi, the etiological agent of Chagas disease, especially in endemic areas.

The complete mitochondrial genome of Choroterpes (Euthralus) yixingensis (Ephemeroptera: Leptophlebiidae) and its mitochondrial protein-coding gene expression under imidacloprid stress.

As one of the most common benthic invertebrates in freshwater, mayflies are very sensitive to changes in water quality and have high requirements for the water environment to allow their nymphs to successfully live and grow. Neonicotinoids, such as imidacloprid, can enter fresh water and pollute the aquatic environment. The present study had two goals: (1) investigate imidacloprid effects on mayfly larvae Choroterpes (Euthralus) yixingensis, and (2) contribute to the phylogenetic status of Ephemeroptera that has always been controversial. Nymphs were collected from Jinhua, China and exposed to different concentrations imidacloprid (5, 10, 20, and 40 µg/L) in the laboratory. Survival of C. yixingensis nymphs decreased as a function of time and imidacloprid concentration with only ~ 55% survival after 72 h exposure to 40 µg/L imidacloprid. After culture under 40 µg/L imidacloprid for 24 h, the steady state transcript levels of mitochondrial COX3, ND4 and ND4L genes were reduced to just 0.07 ± 0.11, 0.30 ± 0.16, and 0.28 ± 0.13 as compared with respective control values (P < 0.01). Steady state transcript levels of ND4 and ND4L were also significantly reduced in a dose-dependent manner (P < 0.05), suggesting that the steady state transcript pattern of these genes in mayfly nymphs can change in response to different levels of environmental contamination. Hence, the mitochondrial protein-coding genes of mayflies could potentially be developed as biomarkers for water ecotoxicity monitoring in the future. In addition, we used the mitochondrial genome sequence of C. yixingensis for an assessment of the phylogenetic tree of Ephemeroptera. The monophyly of Leptophlebiidae was supported and showed that Leptophlebiidae was a sister group to the clade (Baetidae + Caenidae).

Current susceptibilities of brown planthopper Nilaparvata lugens to triflumezopyrim and other frequently used insecticides in China.

The brown planthopper is a notorious rice pest in many areas of Asia. The evolution of insecticide resistance in Nilaparvata lugens has become a serious problem in the effective control of this pest in the paddy field. In this article, the current susceptibility of N. lugens field populations to novel mesoionic insecticide triflumezopyrim and major classes of chemical insecticides was determined and compared. The monitoring results indicated that field populations of N. lugens had developed low resistance to triflumezopyrim (resistance ratio, RR: 1.3-7.3-fold) during 2015-2018 in China, and the median lethal concentration values varied from 0.05 to 0.29 mg/L. Additionally, during 2017 to 2018, field populations of N. lugens showed high resistance levels to thiamethoxam (RR: 456.1-1025.6-fold), imidacloprid (RR: 2195.3-6899.0-fold) and buprofezin (RR: 1241.5-4521.7-fold), moderate to high resistance levels to dinotefuran (RR: 97.6-320.1-fold), clothianidin (RR: 69.4-230.1-fold) and isoprocarb (RR: 44.1-108.0-fold), and low to moderate levels of resistance to chlorpyrifos (RR: 12.0-29.7-fold) and nitenpyram (RR: 6.9-24.1-fold). In contrast, N. lugens just showed low resistance to sulfoxaflor (RR: 3.3-8.5-fold) and etofenprox (RR: 5.0-9.1-fold) in the field. Additionally, the P450 gene CYP6ER1 was found to be significantly overexpressed in all five field populations of N. lugens collected in 2018 when compared with a laboratory susceptible strain. Our findings will provide useful information to delay the evolution of insecticide resistance in N. lugens.

Do-It-Yourself Tick Control: Granular Gamma-Cyhalothrin Reduces Ixodes scapularis (Acari: Ixodidae) Nymphs in Residential Backyards.

Lyme disease is the most common vector-borne disease in the United States with hotspots in the Northeast and Midwest. Integrated vector control for mosquito-borne disease prevention is often organized at the community level, but tick control is primarily coordinated at the household and individual level. Management of the blacklegged tick, Ixodes scapularis (Say), the vector of the causative agent of Lyme disease in the Midwest and eastern United States in peridomestic environments may be critical as many tick encounters are reported to occur in the yard. Therefore, we assessed the effectiveness of a widely available and low-cost pesticide that targets common lawn pests and is labeled for use against ticks. In June 2019, we evaluated a granular form of gamma-cyhalothrin in a placebo-controlled residential backyard study (n = 90) in two communities in Wisconsin. The product applied by the research team reduced nymphal blacklegged ticks in plots established in the lawn part of the ecotone by 97% one week after application at both communities and by 89-97% three to four weeks postapplication. The proportion of homes with at least one nymphal tick postapplication was significantly lower at acaricide-treated homes and ranged from 4.2 to 29.2% compared with placebo homes where at least one nymphal tick was found at 50-81.5% of homes. These results support the efficacy of a low-cost do-it-yourself strategy for homeowners seeking to reduce blacklegged ticks in the yard.

Wolbachia enhances expression of NlCYP4CE1 in Nilaparvata lugens in response to imidacloprid stress.

The brown planthopper, Nilaparvata lugens, is one of the main insect pests of rice. The N. lugens gene NlCYP4CE1 encodes cytochrome P450 monooxygenase (P450), which is a key enzyme in the metabolism of the insecticide imidacloprid. Previous research has suggested that the expression of NlCYP4CE1 is induced by imidacloprid stress, but the effect of bacterial symbionts on its expression has not been determined. The results of this study show that exposure to subtoxic imidacloprid changed the structure of the bacterial symbiont community in N. lugens. Specifically, the total bacterial content increased, but the bacterial species diversity significantly decreased. Wolbachia accounted for the largest proportion of bacteria in N. lugens; its abundance significantly increased after subtoxic imidacloprid exposure. The transcript level of NlCYP4CE1 was significantly increased by imidacloprid, but this effect was significantly weakened after Wolbachia was cleared with tetracycline. This result suggests that Wolbachia enhances the expression of NlCYP4CE1 to promote the detoxification metabolic response to imidacloprid stress. Understanding the effect of bacterial symbionts on gene expression in the host provides a new perspective on interactions between insecticides and their target insect pests, and highlights that subtoxic imidacloprid exposure may raise the risk of insecticide resistance by altering the structure of bacterial symbiont communities.

Blood deprivation and heat stress increase mortality in bed bugs (Cimex lectularius) exposed to insect pathogenic fungi or desiccant dust.

Bed bugs (Cimex lectularius L.) have returned as a nuisance pest in the last 20 years. Different bed bug control measures in combination have not been thoroughly studied, although induction of multiple stressors may improve extermination. The effects of heat stress only, heat stress followed by exposure to insect pathogenic fungi, and heat stress followed by exposure to desiccant dust on starved and blood-fed bed bugs were investigated. Five days at 22 °C (control), 32 °C, 34 °C, or 36 °C (heat stress) did not cause mortality in adults. However, their starved first instar nymphs produced after heat stress suffered mortalities of 33%, 56% and 100%, respectively. Exposure to insect pathogenic fungi after heat stress increased the mortality of adults and their progeny compared to exposure to fungi without heat stress. The beneficial effects of heat stress were not observed in blood-fed bed bugs. Desiccant dust killed all nymphs within 2 days and all adults within 3 days regardless of previous heat stress, but survival time was prolonged by access to blood. This study highlights the advantage of combining different methods in pest management, and points to heat stress combined with blood deprivation as possible management elements to increase the control success.

Synthetic Pyrethroid, Natural Product, and Entomopathogenic Fungal Acaricide Product Formulations for Sustained Early Season Suppression of Host-Seeking Ixodes scapularis (Acari: Ixodidae) and Amblyomma americanum Nymphs.

We compared the ability of product formulations representing a synthetic pyrethroid acaricide (Talstar P Professional Insecticide), a natural product-based acaricide (Essentria IC3), and an entomopathogenic fungal acaricide (Met52 EC Bioinsecticide) to suppress Ixodes scapularis Say and Amblyomma americanum (L.) nymphs when applied following USEPA approved manufacturers' label recommendations for tick control using hand-pumped knapsack sprayers before the beginning of their seasonal activity period in the spring. We applied Met52 EC Bioinsecticide (11% Metarhizium anisopliae Strain F52) to five 100 m2 plots (10.6 ml AI/plot) in mid-April 2020. Two weeks later at the end of April 2020, we treated an additional five 100 m2 plots each with either Talstar P Professional Insecticide (7.9% bifenthrin @ 2.5 ml AI/plot) or Essentria IC3 (10% rosemary oil, 5% geraniol, and 2% peppermint oil @ 86.6 ml AI/plot). Weekly sampling of all plots through the end of June 2020 showed that both Met52 EC Bioinsecticide and Essentria IC3 failed to maintain a 90% suppression threshold for I. scapularis, compared to control plots, and required two additional applications over the course of the trial. In contrast, Talstar P Professional Insecticide suppressed 100% of I. scapularis nymphs and ≥96 and 100% of A. americanum nymphs and adults, respectively. Such pre-season applications of synthetic pyrethroids significantly reduce the early season acarological risk for exposure to host-seeking ticks as well as the frequency of acaricide applications.

Aerobic Metabolism Alterations as an Evidence of Underlying Deltamethrin Resistance Mechanisms in Triatoma infestans (Hemiptera: Reduviidae).

Triatoma infestans (Klug, 1834), the main vector of Chagas disease in Latin America, is regularly controlled by spraying the pyrethroid deltamethrin, to which some populations have developed resistance. The three main mechanisms of resistance are 1) metabolic resistance by overexpression or increased activity of detoxifying enzymes, 2) target site mutations, and 3) cuticle thickening/modification. We use open-flow respirometry to measure real-time H2O loss rate (V˙H2O) and CO2 production rate (V˙CO2), on nymphs from susceptible and resistant populations before and after exposure to the insecticide to understand the underlying mechanisms of resistance in live insects. Lack of differences in V˙H2O between populations suggested that cuticular thickness/composition is not acting as a relevant resistance mechanism. Similarly, there was no difference in resting V˙CO2, suggesting a trade-off between resistance mechanisms and other physiological processes. The increment in V˙CO2 after application of deltamethrin was similar in both populations, which suggested that while enhanced enzymatic detoxification may play a role in resistance expression in this population, the main mechanism involved should be a passive one such as target site mutations. Open-flow respirometry provided useful evidence for evaluating the mechanisms involved in deltamethrin resistance. Using this technique could improve efficiency of scientific research in the area of insecticide resistance management, leading to a faster decision making and hence improved control results.

An efficient high-welfare feeding device for assessing northern fowl mite interventions in vivo: an improved method for the identification of protective antigens/systemic acaricides/repellent effect.

The northern fowl mite (NFM), Ornithonyssus sylviarum, is an obligate hematophagous ectoparasite of domestic and wild birds, and it is an economic pest of laying hen in North America, China, India, Australia, Myanmar, and Brazil. Such an economically important pest remains neglected in many parts of the world, including Asian countries. Therefore, concerted action is required in both basic and applied research directed at the biology and control of this destructive pest. In the present study, we have developed a novel, high-welfare in vivo feeding capsule that would permit pre-screening of new interventions, repellency and deterrence effects of plant-derived products and other semiochemical compounds before proceeding to large-scale field experiments/bioassays, while the minimum number of animals is required to obtain results. Mites were fed on the birds through either a mesh or without a mesh. The average feeding rates of mites was significantly higher when fed directly on chickens, whereas 106 µm nylon mesh was the top-performing mesh when compared with 125 µm aperture nylon mesh. For optimal feeding, the feeding capsules contain NFM and are attached to the skin of the chicken's thigh for 6 h. This is a simple, reproducible, and easy approach and can be adapted to facilitate many aspects of bioassays.

Botanical monoterpenes synergise the toxicity of azamethiphos in the vector of Chagas disease, Triatoma infestans (Hemiptera: Reduviidae).

OBJECTIVE: To investigate what toxicological interactions occur when binary combinations of azamethiphos and botanical monoterpenes (eugenol, menthol or menthyl acetate) are applied to Triatoma infestans. METHODS: The toxicity of binary mixtures of azamethiphos and sublethal doses of a monoterpene (eugenol, menthol or menthyl acetate) was evaluated in nymphs of the first stage of T. infestans. Experiments using exposure to filter papers and topical application were carried out. Values of Lethal Concentration 50% (LC50) were calculated in the first case, and values of Lethal Dose 50% (LD50) in the second. RESULTS: The LC50 of azamethiphos applied on filter paper was 50.3 µg/cm2 . However, when it was simultaneously applied with a sublethal concentration of monoterpene, its toxicity increased (LC50 with eugenol = 11.20 µg/cm2 , LC50 with menthyl acetate = 5.30 µg/cm2 , LC50 with menthol = 7.26 µg/cm2 ). When applied topically, the LD50 of azamethiphos was 7.85 µg/insect, but its toxicity drastically increased when it was applied together with sublethal doses of menthol (LD50 = 0.00016 µg/insect) or menthyl acetate (LD50 = 0.00051 µg/insect). The simultaneous application with eugenol did not significantly change azamethiphos toxicity (LD50 = 12.79 µg/insect). CONCLUSIONS: The toxicity of azamethiphos in T. infestans was synergised when it was applied together with eugenol, menthol or menthyl acetate on a filter paper. However, only menthol and menthyl acetate synergysed azamethiphos when mixtures were topically applied. The drastic effects of menthol and menthyl acetate in topical application experiments should be further studied as they could be the basis for developing more efficient triatomicidal products with a lower content of conventional insecticides than those currently used for controlling T. infestans.

OBJECTIF: Etudier les interactions toxicologiques qui se produisent lorsque des combinaisons binaires d'azaméthiphos et de monoterpènes botaniques (eugénol, menthol ou acétate de menthyle) sont appliquées à Triatoma infestans. MÉTHODES: La toxicité de mélanges binaires d'azaméthiphos et de doses sublétales d'un monoterpène (eugénol, menthol ou acétate de menthyle) a été évaluée sur les nymphes du premier stade de T. infestans. Des expériences utilisant une exposition à des papiers filtres et une application topique ont été réalisées. Les valeurs de concentration létale à 50% (CL50) ont été calculées dans le premier cas et les valeurs de dose létale à 50% (DL50) dans le second. RÉSULTATS: La CL50 de l'azaméthiphos appliqué sur papier filtre était de 50,3 µg/cm2 . Cependant, lorsqu'il était appliqué simultanément avec une concentration sublétale de monoterpène, sa toxicité augmentait (CL50 avec eugénol = 11,20 µg/cm2 , CL50 avec acétate de menthyle = 5,30 µg/cm2 , CL50 avec menthol = 7,26 µg/cm2 ). Lorsqu'il était appliqué localement, la DL50 de l'azaméthiphos était de 7,85 µg/insecte, mais sa toxicité augmentait considérablement lorsqu'il était appliqué avec des doses sublétales de menthol (DL50 = 0,00016 µg/insecte) ou d' acétate de menthyle (DL50 = 0,00051 µg/insecte). L'application simultanée d'eugénol n'a pas modifié de manière significative la toxicité de l'azaméthiphos (DL50 = 12,79 µg/insecte). CONCLUSIONS: La toxicité de l'azaméthiphos chez T. infestans a été mise en synergie lorsqu'il a été appliqué avec de l'eugénol, du menthol ou de l' acétate de menthyle sur un papier filtre. Cependant, seuls le menthol et l' acétate de menthyle ont eu un effet synergique avec l'azaméthiphos lorsque les mélanges étaient appliqués localement. Les effets drastiques du menthol et de l' acétate de menthyle dans les expériences d'application topique devraient être plus étudiés car ils pourraient être la base du développement de produits triatomicides plus efficaces avec une teneur inférieure en insecticides conventionnels que ceux actuellement utilisés pour lutter contre T. infestans.

First report of the lethal activity and synergism between deltamethrin, amitraz and piperonyl butoxide against susceptible and pyrethroid-resistant nymphs of Triatoma infestans.

In South America, Triatoma infestans (Hemiptera: Reduviidae) is the main vector of the parasite Trypanosoma cruzi, etiological agent of Chagas disease. The main strategy for vector control is to spray domestic structures with pyrethroids. Reports of populations of T. infestans with varying degrees of resistance to pyrethroids have made the search for alternative molecules for vector control necessary. In the first stage of this work we investigated the lethal activity of amitraz and deltamethrin against susceptible and pyrethroid-resistant nymphs of Triatoma infestans. Lethal dose at 50% (LD50) of susceptible nymphs were compared with those recorded in pyrethroid-resistant nymphs and the resistance ratio (RR50) was obtained. The RR50 of deltamethrin was approximately 300. In the case of amitraz, we observed similar triatomicidal activity in the two nymph populations (RR50: 0.7). In a second stage of the work, we determined the synergistic effect of amitraz and piperonyl butoxide (PBO) on the lethal activity of deltamethrin. The strong synergistic effect of PBO on the lethal activity of deltamethrin in resistant nymphs produced a decrease in RR50 to almost one third of the RR50 reported in absence of the synergist. Amitraz plus PBO lethal activity was similarly increased in pyrethroid susceptible and resistant nymphs. Our data indicate that deltamethrin synergism by amitraz was higher against resistant than to susceptible nymphs (Synergist ratio (SR50) of: 7.2- and 4.1-fold, respectively). In pyrethroid resistant nymphs, the highest level of synergism was obtained combining deltamethrin with amitraz and PBO (SR50: 26.7-fold). These results indicate that this combination could be considered an effective alternative for the control of T. infestans.

Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata / Determinación de concentraciones letales y niveles de enzimas metabólicas de insecticidas en Triatoma dimidiata

Abstract: Objective: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. Materials and methods: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. Results: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and β-eterase levels. Conclusion: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

Resumen: Objetivo: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. Material y métodos: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. Resultados: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y β-eterasas. Conclusión: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

Respiration, predatory behavior and prey consumption by Podisus nigrispinus (Heteroptera: Pentatomidae) nymphs exposed to some insecticides.

Podisus nigrispinus Dallas (Heteroptera: Pentatomidae) preys on insect pests in eucalyptus plantations where it can be exposed to insecticides used in pest control. The effect of insecticides on non-target natural enemies requires further study. The objective of the present study was to evaluate the side-effects of Bacillus thuringiensis (Bt), permethrin, tebufenozide and thiamethoxam on third instar nymphs of the predator P. nigrispinus in the laboratory. The toxicity of insecticides for this insect was determined by estimating their lethal concentrations. Podisus nigrispinus behavior after exposure to insecticides was analyzed using a video tracking system and the respiratory rate with a respirometer. Prey/nymph consumption was assessed after 24 h of starvation. The preference of P. nigrispinus nymphs, for prey treated or not with the insecticides, was evaluated in free choice tests. The insecticides Bt [LC50 = 1.10(0.83-1.46) mg mL-1], permethrin [LC50 = 0.25(0.17-0.34) mg mL-1], tebufenozide [LC50 = 5.71(4.17-7.57) mg mL-1] and thiamethoxam [LC50 = 0.04(0.02-0.06) mg mL-1] are toxic to P. nigrispinus nymphs. Bt and the insecticides tebufenozide, permethrin and thiamethoxam reduced the respiratory rate of P. nigrispinus. The insecticides permethrin, tebufenozide and thiamethoxam affect the locomotion of this insect's nymphs. Prey treated with Bt, permethrin and thiamethoxam are less preferred by P. nigrispinus. The survival of the nymphs of this predator was 93.3%, 66.7%, 56.6%, 0% and 0% in the control, tebufenozide, Bt, permethrin and thiamethoxam treatments, respectively. In addition, the reduction of prey consumption, treated with neurotoxic insecticides, reduces the predatory potential of this natural enemy. Bt and tefubenozide present low toxicity for P. nigrispinus, but the neurotoxic products have low compatibility with this natural enemy and, therefore, are not recommended, with this predator in the management of forest insect pests.

Molecular screening of cattle ticks, tick-borne pathogens and amitraz resistance in ticks of Santo Domingo de los Tsáchilas province in Ecuador.

The province of Santo Domingo de los Tsáchilas in Ecuador is a strategic place for cattle production and trade. The lack of knowledge about tick species, tick control and tick-borne diseases in Ecuador prompted this study with the goal of identifying the cattle-tick species and tick-borne agents present in the area and molecularly determining the potential acaricide resistance to amitraz of the major cattle tick species. Eighty-four cattle farms were visited and in 88 % of them, cattle were infested with ticks. Additionally, 24 historical samples from other surrounding Ecuadorian provinces, were screened as well. Besides morphological keys, PCR-RFLP MspI was used to confirm the presence of the Rhipicephalus ticks. The tick samples were also screened for tick-borne agents using PCR-RFLP BseDI and Hhal tests to identify circulating Babesia sp. and Anaplasma spp. Furthermore, the PCR-RFLP EciI technique was used to identify the amitraz resistance gene in populations of Rhipicephalus microplus in the province. Pooled testing was used to determine prevalence at individual-tick level. The presence of R. microplus and Amblyomma cajennense sensu lato (s.l.) ticks was found in 83 % and 21 % of the cattle farms respectively, showing R. microplus is widespread in the province of Santo Domingo de los Tsáchilas. Regarding tick-borne agents, only Anaplasma marginale was observed in 50 % of the visited farms of the province, while about 27 % of the ticks tested positive according to estimations from the data of the tick pools. The presence of Babesia bigemina was only confirmed in samples collected outside the province. The amitraz resistance allele in R. microplus was found in 62 % of the farms, but the percentage of farms with cattle ticks completely resistant to this acaricide was low (2%). The findings of this study should prompt cattle producers and animal health authorities to monitor control strategies, which address the management of resistant tick populations and the epidemiologically-unstable areas of tick-borne diseases.

Ultrasound-assisted extraction extracts from Stemona japonica (Blume) Miq. and Cnidium monnieri (L.) Cuss. could be used as potential Rhipicephalus sanguineus control agents.

Nicotiana tabacum, Stemona japonica, and Cnidium monnieri are common plants that are widely used for their anti-parasitic properties. The purpose of this study was to evaluate the acaricidal activity of extracts from these plants against the brown dog tick, Rhipicephalus sanguineus. A composition analysis of crude extracts by GC-MS was conducted to discover compounds with acaricidal effects. The toxicity of extraction against the engorged nymphs of R. sanguineus was evaluated by an immersion test. The results showed that the crude extracts of S. japonica and C. monnieri in varying ratios, concentrations, and from different extraction methods, had a killing effect on R. sanguineus. Lethality reached 76.67% ± 0.04410 when using a 1:1 extract of S. japonica:C. monnieri in 75% ethanol with ultrasonic extraction; the crude extract was determined at a concentration of 0.5 g/mL. GC-MS results showed that osthole and 5-hydroxymethylfurfural (5-HMF) are the main components of the extract. These results suggested that ultrasound-assisted extraction (UAE) extracts contained acaricidal components acting against R. sanguineus, which may result in the development of effective extracts of S. japonica and C. monnieri as a source of low-toxicity, plant-based, natural acaricidal drugs.

Mode of entry of secondary metabolites of the bacteria Xenorhabdus szentirmaii and X. nematophila into Tetranychus urticae, and their toxicity to the predatory mites Phytoseiulus persimilis and Neoseiulus californicus.

The bacterial metabolites in supernatants of Xenorhabdus species have acaricidal activity, but this mode of entry into mites has not yet been elucidated. Herein, we report on the possible mode of entry of Xenorhabdus szentirmaii and Xenorhabdus nematophila supernatants into Tetranychus urticae (Acari: Tetranychidae) adult females. We also assessed the toxicity of the supernatants against the developmental stages of the predatory mites, Phytoseiulus persimilis and Neoseiulus californicus (Acari: Phytoseiidae). Experiments were conducted at 25 ± 1 °C, 70 ± 5% relative humidity, and 16:8h light:dark conditions. Our data showed that the bioactive acaricidal compound is most effective (86.5 to 89% mortality) when the entire integument of T. urticae comes in contact with it compared to contact of the ventral side only (26.5-34%). Against P. persimilis and N. californicus at 6 days post-application (dpa), the eggs were not affected by the X. szentirmaii or X. nematophila supernatant, whereas mortality of the mobile stages (larva, protonymph, deutonymph, adult) was 18.5% to 39.2%. Overall, the predatory mites were less affected by the bacterial metabolites than T. urticae. We hypothesize that the differences in morphology such as longer legs and thicker cuticle, as well as the diet of the predatory mites, reduce the contact of the body parts to the supernatant-treated surfaces. We need to isolate, identify, and characterize the X. szentirmaii and X. nematophila metabolite(s) and demonstrate efficacy to pestiferous mites and safety to plants, non-target organisms and the environment before it can be used as an acaricide.