Effects of Terbufos (Organophosphate) on Larval Behaviour of Two Forensically Important Diptera Species: Contributions for Entomotoxicology.

Neurotoxicant compounds interfere with the behaviour and biology of insects, significantly altering their locomotion patterns. However, little is known about the effect of organophosphates, neurotoxicants for agricultural, domestic and industrial use, on the larval movement of necrophagous flies, although being responsible for frequent cases of poisoning and accidental or intentional deaths. Thus, we aimed to study the influence of Terbufos (organophosphate) on the activity and mobility patterns of Lucilia eximia (Wiedemann 1819) (Calliphoridae) and Peckia (Peckia) chrysostoma (Wiedemann 1830) (Sarcophagidae) immatures collected from rat carcasses intoxicated with 5, 10 or 20 mg/kg of Terbufos, to evaluate (i) peristaltic movements and body contractions, and (ii) distance and shape of the trajectory travelled by the larva. Behavioural parameters were analysed in loco and through videos. We observed that the presence of Terbufos altered poisoned larvae's activity and body mobility in both taxon and dose-dependent manner. Lucilia eximia larvae were more active, with greater frequency of body movements and lateral contractions when intoxicated with high and intermediate doses of Terbufos. On the other hand, P. (P.) chrysostoma immatures were less active, with fewer body and lateral contractions when intoxicated with the high dose of the compound. This work experimentally demonstrates that the presence of Terbufos can alters the mobility and movement of intoxicated necrophagous Diptera, essential components of the cadaveric fauna.

The repellency effect of icaridin nanostructural solution applied on cotton knitting fabric against Lutzomyia longipalpis.

The use of repellents is considered an alternative against biting insects, including Lutzomyia longipalpis (Diptera: Psychodidae), the main vector of the protozoan Leishmania infantum, visceral leishmaniasis's (VL) etiologic agent in the Americas. This study aimed to evaluate the repellent efficacy of icaridin nanostructured solution applied on cotton knitting fabric against L. longipalpis. Arm-in-cage tests were performed in eight volunteers at different concentrations (5%, 10%, 25%, and 50%), using L. longipalpis (n = 30). The bioassay was performed in 1, 24, 48, 72, 96, 120, and 144 h after impregnation and one test after washing the fabrics with icaridin. The total repellency rate (%R) > 95% was used as a reference to define a minimum effective concentration (MEC). The results revealed that the insects' landing mean decreased significantly in different icaridin concentrations, compared with the control tests (p < 0.05) and the 25% and 50% concentrations compared to lower concentration (5%) (p < 0.05). The higher concentrations (25% and 50%) provided longer complete protection times (CPTs) with 120 and 144 h of protection, respectively and the %R of 100% for 72 and 96 h after impregnation, respectively. The 25% was the MEC (%R Total = 98.18%). Our results indicate, for the first time, that icaridin nanostructured solution applied on cotton knitting fabric proved to be an efficient repellent against L. longipalpis with the presence of repellent action even after washing. The concentration of 25% showed better efficiency and may become an efficient method for L. longipalpis biting control.

Culicidae (Diptera: Culicomorpha) in the southern Brazilian 'Ana Leuch Lozovei' collection, with notes on distribution and diversity.

Biological collections are important for acquiring knowledge of the biodiversity of a specific environment. Here, we organize, list and catalog adult specimens of Culicidae from the Parasitology Collection of the Basic Pathology Department at the Federal University of Paraná, Brazil, and present a databank of taxonomic and collection information for each sample. Culicids were collected using different methodologies in 18 municipalities in Paraná state, between 1967 and 1999. There are 5,739 catalogued specimens, of which 4,703 (81.94%) are identified at species level, with a diversity of 100 species. Of these, 18 are new recorded samples for Paraná, and three are new for Brazil. This collection is named the Ana Leuch Lozovei Entomological Collection in honor of the 30 years Prof Lovozei dedicated to culicid study in Paraná. The collection comprises insect vectors or potential vectors, of agents causing human diseases.

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.

Behavioural Aspects of the Prey-Predator Interaction Among Necrophagous Diptera: Implications for Cadaveric Colonization.

Predation is a major process in determining the composition and dynamics of necrophagous dipteran assemblages and has implications for the colonization of cadavers. This work describes behavioural interactions between necrophagous larvae under a predator-prey perspective, using as a model the predatory species Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae) and three species of prey: Lucilia eximia (Wiedemann) (Calliphoridae), Synthesiomyia nudiseta (Wulp) (Muscidae) and Peckia (Peckia) chrysostoma (Wiedemann) (Sarcophagidae). Using experimental arenas, we observed behavioural acts, such as escape, avoidance and prey discrimination in three experiments, which also incorporated predator density. We demonstrate that the number of escapes, avoidance and predation varies according to the type of prey. Lucilia eximia and P. chrysostoma are able to avoid and/or escape the predator more frequently when compared to S. nudiseta. At least 70% of larvae of both species successfully evaded from C. albiceps attack. Low frequency of escape, associated with strong ability to counterattack, was observed for S. nudiseta. Peckia chrysostoma and L. eximia were the preferred items in the experiment of prey discrimination in both 2- and 3-choice tests. Predation occurs quickly, and the minimum predation time varied according to the prey species and predator density. Remarkably, prey handling varied with the species: whilst individual larvae chased and killed L. eximia, 75% of predation of P. chrysostoma occurred in groups of two or more C. albiceps larvae. Differential intraguild predation has implications for reducing populations of carrion insects-consequently affecting the likelihood of sampling necrophagous larvae in human cadavers.

Fluralaner (Bravecto®) induces long-term mortality of Lutzomyia longipalpis after a blood meal in treated dogs.

BACKGROUND: Leishmania infantum is the etiological agent of visceral leishmaniasis (VL) in the New World, where the sand fly Lutzomyia longipalpis and domestic dogs are considered the main vector and host reservoirs, respectively. Systemic insecticides have been studied as an alternative to control vector-borne diseases, including VL. Fluralaner, an isoxazoline class compound, is a systemic insecticide used in dogs, with proven efficiency against different species of phlebotomine sand flies. However, to date no studies have demonstrated the efficacy of fluralaner on Lu. longipalpis. The aim of this study was to evaluate the insecticidal effect of fluralaner (Bravecto®) on the sand fly Lu. longipalpis after blood meal in treated dogs. METHODS: Healthy mongrel dogs (n = 8) were recruited from the Zoonoses Control Center in the city of Natal, Rio Grande do Norte, Brazil, and randomized into two groups: fluralaner treated (n = 4) and non-treated control (n = 4). Colony-reared female specimens of Lu. longipalpis (n = 20) were allowed to feed on all dogs for 40 min before treatment (for fluralaner-treated dogs), at day 1 after treatment and then monthly until 1 year post-treatment. RESULTS: In the treatment group, there was 100% mortality of Lu. longipalpis for up to 5 months after treatment initiation, decreasing to 72.5% at 6 months post-treatment initiation. The efficacy of fluralaner ranged from 100% at day 1 (P = 0.0002) to 68% ( P = 0.0015) at 6 months, decreasing to 1.4% at 1 year post-treatment. Sand fly mortality carried out blood meal in non-treated control dogs remained constant at ≤ 15%. CONCLUSIONS: Taken together, our results suggest that fluralaner may be used as a control strategy for VL in dogs in VL endemic areas.

Carrion decomposition and assemblage of necrophagous dipterans associated with Terbufos (Organophosphate) intoxicated rat carcasses.

Organophosphate use represents one of the main causes of intentional and accidental poisoning in Brazil. Because they induce alteration in carcass decomposition and succession of necrophagous Diptera, they are important investigative tool in forensic entomology for determining the postmortem interval (PMI) and cause of death. Thus, this work aimed to evaluate the influence of Terbufos, the most commonly used organophosphate for suicides in Rio Grande do Norte, Brazil, in the decomposition process and necrophagous fly assemblies in intoxicated rat carcasses. For this, 150g female Wistar rats received, by gavage, 200µL of Terbufos (5mg/kg or 10mg/kg) or distilled water (control). Approximately 30 minutes after administration, animals were euthanized and distributed in suspended traps to decompose under environmental conditions. Decomposition was monitored daily, with photographic record and collection of the visiting dipterofauna until the dry phase. Data show that the higher dose of Terbufos i) accelerates carcass decomposition in 24h; ii) decreases the species richness and abundance of scavengers flies when compared to the control; iii) changes the succession pattern, delaying the arrival of important species for the PMI estimate and iv) causes 8% mortality of the visiting dipterofauna. This work provides relevant information about dipteran assemblages and changes in the cadaver decomposition process associated with Terbufos intoxication, which may assist in future investigative processes with suspected organophosphate poisoning.

Diversity of Mosquitoes (Diptera: Culicidae) in the Caatinga Biome, Brazil, from the Widespread to the Endemic.

Mosquito fauna in the northeast semiarid region of Brazil, Caatinga biome, are poorly known. Studies on the diversity are scarce and the few surveys available focus on local fauna. In order to understand the ecological pattern of mosquito's distribution, information available from studies from 2008 to 2015 were gathered. A partitioning framework of the beta diversity, the turnover (ßJTU) and nestedness (ßJNE) components were used to determine dissimilarity among communities. Eighty-two morphospecies were recorded and 47 of the species were not shared between the areas. The most representative genera were Aedes, Anopheles, Psorophora, Haemagogus, Coquillettidia, and Mansonia, which all include species of medical interest. The communities had high rates of variation, and the mechanism of turnover accounted for the observed diversity pattern. Despite differences in collection methods, the observed dissimilarity may be related to the broad environmental heterogeneity of the biome, the intrinsic relationships of the species with their habitats, and the environmental degradation caused by different types of anthropogenic interference. Considering the mosquito species richness and endemicity, the hypothesis that the Caatinga harbor poor biodiversity is rejected. The spatial variation observed is of particular importance and should be taken into account for the knowledge of Caatinga biodiversity.

The Effect of Flunitrazepam (Rohypnol(®) ) on the Development of Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) and its Implications for Forensic Entomology.

This study investigated the potential effects of flunitrazepam (known as "date rape drug") on the developmental cycle of Chrysomya megacephala, an important forensic species, and their possible implications for the calculation of the PMI. A 1050 C. megacephala eggs were divided into five groups with seven replications each. The eggs were placed on artificial diet prepared with four drug concentrations of flunitrazepam (4, 8, 16, and 32 ng/g), besides the control group (prepared with water). Were evaluated the potential effects on development time, weight gain, and mortality during the cycles. The drug had no significant effect on development time or mortality although it did affect the weight of the pupae and adults (Kruskal-Wallis, p < 0.05). The result can be deduced that the determination of the postmortem interval is not affected.

Development of the BG-Malaria trap as an alternative to human-landing catches for the capture of Anopheles darlingi.

Although the human-landing catch (HLC) method is the most effective for collecting anthropophilic anophelines, it has been increasingly abandoned, primarily for ethical considerations. The objective of the present study was to develop a new trap for the collection of Anopheles darlingi . The initial trials were conducted using the BG-Sentinel trap as a standard for further trap development based on colour, airflow direction and illumination. The performance of the trap was then compared with those of the CDC, Fay-Prince, counterflow geometry trap (CFG) and HLC. All trials were conducted outdoors between 06:00 pm-08:00 pm. Female specimens of An. darlingi were dissected to determine their parity. A total of 8,334 anophelines were captured, of which 4,945 were identified as An. darlingi . The best trap configuration was an all-white version, with an upward airflow and no required light source. This configuration was subsequently named BG-Malaria (BGM). The BGM captured significantly more anophelines than any of the other traps tested and was similar to HLC with respect to the number and parity of anophelines. The BGM trap can be used as an alternative to HLC for collecting anophelines.

Development of the BG-Malaria trap as an alternative to human-landing catches for the capture of Anopheles darlingi

Although the human-landing catch (HLC) method is the most effective for collecting anthropophilic anophelines, it has been increasingly abandoned, primarily for ethical considerations. The objective of the present study was to develop a new trap for the collection of Anopheles darlingi . The initial trials were conducted using the BG-Sentinel trap as a standard for further trap development based on colour, airflow direction and illumination. The performance of the trap was then compared with those of the CDC, Fay-Prince, counterflow geometry trap (CFG) and HLC. All trials were conducted outdoors between 06:00 pm-08:00 pm. Female specimens of An. darlingi were dissected to determine their parity. A total of 8,334 anophelines were captured, of which 4,945 were identified as An. darlingi . The best trap configuration was an all-white version, with an upward airflow and no required light source. This configuration was subsequently named BG-Malaria (BGM). The BGM captured significantly more anophelines than any of the other traps tested and was similar to HLC with respect to the number and parity of anophelines. The BGM trap can be used as an alternative to HLC for collecting anophelines.

Fauna of Culicidae in rural areas of Porto Velho and the first record of Mansonia (Mansonia) flaveola (Coquillet, 1906), for the state of Rondônia, Brazil.

INTRODUCTION: Knowledge concerning the fauna of Culicidae in the Brazilian Amazon States contributes to current understanding of the bionomics of the insects collected and makes it possible to observe changes in the fauna over time. METHODS: The Culicidae were captured with a BG-Sentinel® trap in extra-domiciliary area of two rural regions of Porto Velho in June and July of 2007 and 2008. RESULTS: A total of 10,695 Culicidae was collected, belonging to nine genera: Coquillettidia, Culex, Mansonia, Psorophora, Aedes, Aedeomyia, Anopheles, Uranotaenia and Wyeomyia. CONCLUSIONS: The presence of Mansonia (Mansonia) flaveola was recorded in the State of Rondônia for the first time.

Fauna of Culicidae in rural areas of Porto Velho and the first record of Mansonia (Mansonia) flaveola (Coquillet, 1906), for the state of Rondônia, Brazil / Fauna de Culicidae em áreas rurais de Porto Velho e primeiro registro de Mansonia (Mansonia) flaveola (Coquillet, 1906), para o estado de Rondônia, Brasil

INTRODUCTION: Knowledge concerning the fauna of Culicidae in the Brazilian Amazon States contributes to current understanding of the bionomics of the insects collected and makes it possible to observe changes in the fauna over time. METHODS: The Culicidae were captured with a BG-Sentinel® trap in extra-domiciliary area of two rural regions of Porto Velho in June and July of 2007 and 2008. RESULTS: A total of 10,695 Culicidae was collected, belonging to nine genera: Coquillettidia, Culex, Mansonia, Psorophora, Aedes, Aedeomyia, Anopheles, Uranotaenia and Wyeomyia. CONCLUSIONS: The presence of Mansonia (Mansonia) flaveola was recorded in the State of Rondônia for the first time.

INTRODUÇÃO: O conhecimento da fauna de culicídeos em estados da Amazônia Brasileira auxilia no conhecimento da bionomia dos insetos obtidos, além de possibilitar a observação de modificações da fauna ao decorrer do tempo. MÉTODOS: As capturas dos culicídeos foram realizadas em área extradomiciliar de duas zonas rurais de Porto Velho, durante junho e julho, nos anos de 2007 e 2008, com a armadilha BG-Sentinel®. RESULTADOs: Foram coletados 10.695 culicídeos, distribuídos em nove gêneros: Coquillettidia, Culex, Mansonia, Psorophora, Aedes, Aedeomyia, Anopheles, Uranotaenia e Wyeomyia. CONCLUSÕES: Foi registrada pela primeira vez no estado a presença de Mansonia (Mansonia) flaveola.

Patent literature on mosquito repellent inventions which contain plant essential oils--a review.

Bites Bites of mosquitoes belonging to the genera Anopheles Meigen, Aedes Meigen, Culex L. and Haemagogus L. are a general nuisance and are responsible for the transmission of important tropical diseases such as malaria, hemorrhagic dengue and yellow fevers and filariasis (elephantiasis). Plants are traditional sources of mosquito repelling essential oils (EOs), glyceridic oils and repellent and synergistic chemicals. A Chemical Abstracts search on mosquito repellent inventions containing plant-derived EOs revealed 144 active patents mostly from Asia. Chinese, Japanese and Korean language patents and those of India (in English) accounted for roughly 3/4 of all patents. Since 1998 patents on EO-containing mosquito repellent inventions have almost doubled about every 4 years. In general, these patents describe repellent compositions for use in topical agents, cosmetic products, incense, fumigants, indoor and outdoor sprays, fibers, textiles among other applications. 67 EOs and 9 glyceridic oils were individually cited in at least 2 patents. Over 1/2 of all patents named just one EO. Citronella [Cymbopogon nardus (L.) Rendle, C.winterianus Jowitt ex Bor] and eucalyptus (Eucalyptus LʼHér. spp.) EOs were each cited in approximately 1/3 of all patents. Camphor [Cinnamomum camphora (L.) J. Presl], cinnamon (Cinnamomum zeylanicum Blume), clove [Syzygium aromaticum (L.) Merr. & L.M. Perry], geranium (Pelargonium graveolens LʼHér.), lavender (Lavandula angustifolia Mill.), lemon [Citrus × limon (L.) Osbeck], lemongrass [Cymbopogon citratus (DC.) Stapf] and peppermint (Mentha × piperita L.) EOs were each cited in > 10% of patents. Repellent chemicals present in EO compositions or added as pure "natural" ingredients such as geraniol, limonene, p-menthane-3,8-diol, nepetalactone and vanillin were described in approximately 40% of all patents. About 25% of EO-containing inventions included or were made to be used with synthetic insect control agents having mosquito repellent properties such as pyrethroids, N,N-diethyl-m-toluamide (DEET), (±)-p-menthane-3,8-diol (PMD) and dialkyl phthalates. Synergistic effects involving one or more EOs and synthetic and/or natural components were claimed in about 10% of all patents. Scientific literature sources provide evidence for the mosquito repellency of many of the EOs and individual chemical components found in EOs used in patented repellent inventions.

Efeito da ovitrampa letal na longevidade de fêmeas de Aedes aegypti (Diptera: Culicidae) / Effect of lethal ovitrap on the longevity of females of Aedes aegypti (Diptera: Culicidae)

A armadilha de oviposição acrescida de inseticida pode funcionar como novo método de controle de fêmeas do mosquito Aedes aegypti. Fêmeas de Aedes aegypti foram colocadas em contato com ovitrampas letais envelhecidas e a mortalidade variou de 60,3 por cento a 100 por cento sendo significativo o efeito do envelhecimento das palhetas impregnadas com deltametrina no percentual de mortalidade.

Oviposition traps with added insecticide may work as a new method for controlling the females of the Aedes aegypti mosquito. Females of Aedes aegypti were placed in contact with lethal ovitraps with aging. The mortality rate ranged from 60.3 percent to 100 percent. The effect of aging the slats impregnated with deltamethrin was significant in relation to the percentage mortality among Aedes aegypti females.

[Evaluation of the HP trap baited with different release rates of octenol for capturing anophelines (Diptera: Culicidae) in Brejo do Mutambal, municipality of Varzelândia, State of Minas Gerais]. / Avaliação da armadilha HP iscada com diferentes taxas de liberação de octenol na captura de anofelinos (Diptera: Culicidae) em Brejo do Mutambal, Município de Varzelândia, Estado de Minas Gerais.

To evaluate the role of octenol in collecting anophelines, HP traps baited with different rates of evaporation. A total of 285 anophelines was captured and there was no significant difference in the numbers of anophelines captured between the control traps and the traps baited with octenol and no difference between the release rates.

Avaliação da armadilha HP iscada com diferentes taxas de liberação de octenol na captura de anofelinos (Diptera: Culicidae) em Brejo do Mutambal, Município de Varzelândia, Estado de Minas Gerais / Evaluation of the HP trap baited with different release rates of octenol for capturing anophelines (Diptera: Culicidae) in Brejo do Mutambal, municipality of Varzelândia, State of Minas Gerais

Para avaliar o papel do octenol na coleta de anofelinos foram utilizadas armadilhas HP iscadas com diferentes taxas de evaporação. Foram capturados 285 anofelinos e não houve diferença entre o número de anofelinos coletados com as armadilhas controle e as iscadas com octenol e nem entre as taxas de evaporação.

To evaluate the role of octenol in collecting anophelines, HP traps baited with different rates of evaporation. A total of 285 anophelines was captured and there was no significant difference in the numbers of anophelines captured between the control traps and the traps baited with octenol and no difference between the release rates.

[Effect of lethal ovitrap on the longevity of females of Aedes aegypti (Diptera: Culicidae)]. / Efeito da ovitrampa letal na longevidade de fêmeas de Aedes aegypti (Diptera: Culicidae).

Oviposition traps with added insecticide may work as a new method for controlling the females of the Aedes aegypti mosquito. Females of Aedes aegypti were placed in contact with lethal ovitraps with aging. The mortality rate ranged from 60.3% to 100%. The effect of aging the slats impregnated with deltamethrin was significant in relation to the percentage mortality among Aedes aegypti females.

[Persistence and efficacy of growth regulator pyriproxyfen in laboratory conditions for Aedes aegypti]. / Persistência e eficácia do regulador de crescimento pyriproxyfen em condições de laboratório para Aedes aegypti.

The persistence and efficacy of growth regulator pyriproxyfen were evaluated in two final concentrations 0.01 and 0.05 ppm against Aedes aegypti larvae in laboratory conditions using three types of containers: cement box (45 liters), glass bottle (5 liters) and plastic bucket (20 liters). The tests were carried after 1, 7, 15, 30, 45, 60, 90 and 120 days of treatment against Aedes aegypti larvae 3rd and 4th instar. The percentages of larvae, pupae and adult mortality, the percentage of adult emergence inhibition and time duration of bioassays were calculated. A was observed a persistence of 45 and 90 days by using 0.01 and 0.05 ppm final concentrations of pyriproxyfen, respectively, was observed. We observed that mortality in the pupa stage was significantly higher than larvae and adults mortality for all containers and concentrations.

Persistência e eficácia do regulador de crescimento pyriproxyfen em condicões de laboratório para Aedes aegypti / Persistence and efficacy of growth regulator pyriproxyfen in laboratory conditions for Aedes aegypti

A persistência e a eficácia do regulador de crescimento pyriproxyfen foram testadas em concentracões de 0,01 e 0,05ppm, contra larvas de Aedes aegypti, utilizando os recipientes caixas d'água (45 litros), frascos de vidro (5 litros) e baldes de plástico (20 litros). As avaliacões foram nos dias 1, 7, 15, 30, 45, 60, 90 e 120 após o tratamento usando larvas de 3º e 4º estádio de Aedes aegypti. Foi calculado o percentual de mortalidade de larvas, pupas e adultos, percentual de inibicão de emergência de adulto e duracão dos bioensaios. Observou-se que a persistência foi de 45 dias e 90 dias para concentracão final de 0,01 e 0,05ppm de pyriproxyfen, respectivamente. Observamos que a mortalidade de pupas foi significativamente maior que a de larvas e de adultos para todos os recipientes e concentracões.