Assessment of infection of Stomoxys calcitrans larvae by entomopathogenic nematodes Heterorhabditis amazonensis NEPT11.

This study aimed to evaluate the virulence of Heterorhabditis amazonenses NEPT11 against larvae of Stomoxys calcitrans. Groups of 10 third-instar fly larvae were deposited in Petri dishes, to which were added 50, 100 and 200 EPNs/larva in 4ml of distilled water. The volume of the control group was the same as the treated group, but without EPNs. Larval mortality was observed daily, until larvae died or adults emerged. The Petri dishes were kept on laboratory shelves at 27 ± 1 °C and 70 ± 10% RH. The experiment was replicated six times. A regression analysis revealed quadratic behavior with increasing concentrations, indicating that the concentration of 200 EPNs/larva (48%) was the most efficient among the tested concentrations, while concentrations of 50 and 100 EPNs/larva killed 26.6 and 40% of larvae, respectively. In general, none of the treatments resulted in a mortality rate of more than 50%, but all the treated groups exhibited a higher mortality than that of the control group. It is concluded that the EPN H. amazonensis NEPT11 shows a promising potential to control third-instar larvae of S. calcitrans. However, further studies are needed in different situations to better understand the activity of this organism against the immature stages of the stable fly.

Este estudo teve como objetivo avaliar a ação do NEP Heterorhabditis amazonenses NEPT11 frente larvas de S. calcitrans. Grupos de 10 larvas de terceiro instar da mosca foram depositados em placas de Petri, em seguida, adicionou-se 50, 100, 200, 300 e 400 NEPs/larva em 4ml de água destilada. O volume do grupo controle foi o mesmo dos tratados, porém sem NEPs. A mortalidade das larvas foi observada diariamente, até a morte das larvas ou emergência de adultos. As placas foram mantidas em estantes de laboratório a 27 ± 1 °C e 70 ± 10% UR. O experimento teve seis repetições. Por meio da análise de regressão, foi observado comportamento quadrático com o aumento das concentrações, sendo a concentração de 200 NEPs/larva (48%) a de maior eficiência entre as concentrações testadas, já as concentrações de 50 e 100 NEPs/larva mataram 26,6 e 40% das larvas, respectivamente. De modo geral, nenhum tratamento proporcionou mortalidade superior a 50%, todavia, todos os grupos tratados apresentaram mortalidade superior à observada no controle. Conclui-se que H. amazonenses NEPT11 mostrou-se promissor no controle de larvas de terceiro instar de S. calcitrans, porém mais estudos devem ser feitos para o melhor entendimento da ação deste organismo frente aos estágios imaturos da mosca-dos-estábulos.

Evaluation of mechanical transmission of Trypanosoma vivax by Stomoxys calcitrans in a region without a cyclic vector.

This work investigated the mechanical transmission of Trypanosoma vivax by Stomoxys calcitrans to cattle in a region without a cyclic vector. The study involved two experiments, one with calves experimentally infected with T. vivax, in the acute phase of trypanosomosis (Experiment 1) and the other in the chronic phase (Experiment 2). In both experiments, two transmission methods were used with flies that had not fed for 24 h or had never fed: (i) Method 1: flies released freely in cattle pens (≈3,300 flies/pen for 10 days); and (ii) Method 2: flies placed in a feeding chamber (12 flies/animal). To develop Method 1 in the two experiments (acute and chronic phases), T. vivax-positive animals were kept with T. vivax-negative animals. Periodically, the Brener method, Woo method, blood smears, cPCR, ELISA, IFAT, and Imunoteste® were performed to detect T. vivax in the animals. We also recorded the animals' head tossing and hoof stomping and the number of flies near the pens' inner walls. Subsequently, biological testing was performed using lambs. For Method 2 in both experiments, flies inside the feeding chamber first fed on T. vivax-positive animals and later on negative animals. In both experiments and methods, we examined the flies for the presence of T. vivax through blood smears and cPCR of the proboscis and abdomen. In Experiment 2 (chronic phase), a test was conducted to determine how long trypomastigotes forms could survive on the blood of animals with different levels of parasitemia. None of the animals (calves and lambs) became infected with T. vivax or showed antibodies against it. During the evaluation period, the animals in the presence of the flies exhibited more hoof stomping and head tossing compared to those without flies (control). Additionally, there was an increase in the number of flies in the pens during the experiment. Only in Experiment 1 (acute phase) were T. vivax trypomastigotes and DNA found in the abdomen of the flies but not in the proboscis. In Experiment 2 (chronic phase), higher concentrations of trypomastigotes per milliliter of blood were associated with a shorter the lifespan of this stage of the parasite. In conclusion, under the variable conditions of the experiments (hosts, number of flies, and level of parasitemia), S. calcitrans was unable to mechanically transmit T. vivax to cattle.

Evaluation of the effect of Heterorhabditis bacteriophora (HP88) on Stomoxys calcitrans (Linnaeus, 1758) larvae (Diptera: Muscidae) in sugarcane bagasse ash.

The purpose of this study was to evaluate the effect of the EPN Heterorhabditis bacteriophora HP88 on Stmoxys. calcitrans larvae in sugarcane bagasse ash. Groups of 10 stable fly larvae were placed in Petri dishes containing filter paper and bagasse ash. Concentrations of 50, 150 and 250 EPNs/larva of S. calcitrans in four milliliters of distilled water were added to each plate. In the control group contained only distilled water, without EPNs. The bioassay had three replications and was maintained at 27 ± 1°C and 70-80% relative humidity. It was observed that mortality rate in all treated groups was significantly higher than in the control group (26,6%). The mortality rate in the presence of 50 EPNs/larva (46,6%) was lower than in 150 EPNs/larva (76,3%), which in turn was lower than 250 EPNs/larva group (93,3%). It was verified by analysis of variance and regression that there was a linear pattern of mortality, that is, the higher the EPNs/larva concentration, the higher the larval mortality. It was concluded that EPN H. bacteriophora HP88 was capable of infecting and causing mortality of stable fly larvae in sugarcane bagasse ash.

O objetivo deste estudo foi avaliar o efeito do NEP Heterorhabditis bacteriophora HP88 sobre larvas de Stomoxys calcitrans em cinzas de bagaço de cana-de-açúcar. Grupos de 10 larvas da mosca dos estábulos foram depositadas em placas de Petri contendo papel filtro e cinzas. Foram adicionadas concentrações de 50, 150 e 250 NEPs/larva de S. calcitrans em cada placa. No grupo controle não havia NEPs, somente água destilada. O bioensaio teve três repetições e foi mantido em 27 ± 1°C e 70-80% de umidade relativa. Observou-se que a mortalidade em todos os grupos tratados foi significativamente superior à do grupo controle (26,6%). A taxa de mortalidade na presença de 50 NEPs/larva (46,6%) foi menor do que em 150 NEPs/larva (76,3%), que por sua vez foi menor do que no grupo 250 NEPs/larva (93,3%). Verificou-se pela análise de variância e de regressão que houve um padrão linear de mortalidade, ou seja, quanto maior a concentração de NEPs/larva, maior a mortalidade larval. Conclui-se que o NEP H. bacteriophora HP88 foi capaz de infectar e causar mortalidade das larvas da mosca dos estábulos em cinza de bagaço de cana e que aparentemente este subproduto não interfere negativamente na ação deste NEP.

A survey of Stomoxys Geoffroy, 1762 (Diptera: Muscidae) in eight administrative regions of Cameroon

ABSTRACT Stomoxys Geoffroy, 1762 are major livestock pests in the tropics and are common in diverse habitats. This study aims to conduct a survey on the Stomoxys fauna of Cameroon. From 2015 to 2017, entomological studies using standard traps (n=204) were conducted in eight administrative regions found in five agro-ecological zones (AEZs) of Cameroon with 606 trap-points over 22,032 traps days. A total of 77,804 Stomoxys specimens were collected, with eight taxa consisting of six species (S. calcitrans (Linnaeus, 1758), S. omega Newstead, Dutton & Todd, 1907, S. xanthomelas Roubaud, 1937, S. inornatus Grunberg, 1906, S. transvittatus Villeneuve, 1916, and S. sitiens (Rondani, 1873)) and two subspecies (Stomoxys niger niger Macquart, 1851 and S. niger bilineatus Grunberg, 1906) identified. Among all the recorded taxa, S. calcitrans and S. n. niger were present in five and seven of the eight regions respectively, but S. sitiens was rare and only found in the North. Furthermore, the highest species number (seven out of eight) was recorded in the Guinee savanna of the Adamawa region. The highest apparent density range of 101 to 200 Stomoxys/ trap/ day (s/t/d) was recorded in the Sudan savanna AEZ of the Far North region. Stomoxys occurred in all the AEZs in eight regions, some of which are major cattle rearing regions. This represents risk for the mechanically transmission of dangerous pathogens in those regions.

Evaluation in vitro of the virulence of two entomopathogenic heterorhabditid nematodes in the control of Stomoxys calcitrans (Diptera: Muscidae) larvae in byproducts of the sugar and alcohol industry / Avaliação in vitro da virulência de dois nematoides entomopatogênicos heterorhabditídeos no controle de larvas de Stomoxys calcitrans (Diptera: Muscidae) em subprodutos da indústria sucroalcooleira

Stomoxys calcitrans causes losses to livestock, mainly to cattle. This study aimed to determine the pathogenic potential of Heterorhabditis bacteriophora HP88 and H. baujardi LPP7 against S. calcitrans larvae after being exposed to byproducts of the sugar and alcohol industry. The efficacy of EPNs on stable fly larvae was evaluated in bioassays with vinasse at three temperatures (16, 25 and 35 °C) and concentrations (0, 50 and 100%), as well as in relation to larva age (4, 6 and 8 days) in filter cake and EPNs concentration (100, 300 and 500 IJs/larva) in sugarcane bagasse. H. bacteriophora showed higher efficacy than H. baujardi at all temperatures. Vinasse did not have a negative effect on the virulence of H. bacteriophora. The age of fly larvae did not affect the mortality rates caused by the EPNs. In bagasse, H. bacteriophora presented higher mortality rates than the control group. It is concluded that EPNs can be a potential component in integrated strategies of stable fly control and outbreak prevention in areas of sugar and alcohol production.(AU)

Stomoxys calcitrans causa perdas à agropecuária, principalmente na bovinocultura. Este estudo objetivou verificar o potencial patogênico de Heterorhabditis bacteriophora HP88 e H. baujardi LPP7 para larvas de S. calcitrans, após serem expostos a subprodutos da indústria sucroalcooleira. A eficácia dos NEPs sobre larvas da mosca-dos-estábulos foi avaliada em bioensaios com vinhaça em três temperaturas (16, 25 e 35 °C) e concentrações (0, 50 e 100%), bem como em relação à idade da larva (4, 6 e 8 dias) em torta de filtro e concentração de NEPs (100, 300 e 500 JIs/larva) em bagaço de cana-de-açúcar. H. bacteriophora apresentou maior eficácia do que H. baujardi em todas as temperaturas. O vinhoto não apresentou efeito negativo sobre a virulência de H. bacteriophora. A idade das larvas não influenciou no aumento ou diminuição da mortalidade dos NEPs. Em bagaço de cana, H. bacteriophora apresentou taxas de mortalidade superiores à do grupo controle. Conclui-se que os NEPs podem ser um potencial componente em estratégias de controle integrado e prevenção de surtos da mosca-dos-estábulos, em áreas de produção de açúcar e álcool.(AU)

First report of the presence of Anaplasma marginale in different tissues of the stable-fly Stomoxys calcitrans (Linnaeus, 1758) in Rio de Janeiro state, Brazil.

Stomoxys calcitrans is a cosmopolitan hematophagous insect with significant veterinary importance. Besides causing great stress with its bites, the fly is a mechanical vector of several pathogens of the most diverse phylogenetic groups to their hosts. The objective of this study was to test for the presence of Anaplasma marginale in stable flies collected at three points inside the campus of the Federal Rural University of Rio de Janeiro (UFRRJ), Seropédica, Rio de Janeiro, Brazil. The collected flies were dissected and separated into three parts - heads, carcass and guts - which were individually submitted to A. marginale specific semi nested PCR gene amplification. A total 150 samples of 50 flies were assayed and 11.33% were positive with predominant presence in guts. In parallel, 6 F1 colony S. calcitrans flies fed with bovine blood were also investigated, being all positive for the presence of the bacteria. This is the first report of the presence of A. marginale in S. calcitrans at Rio de Janeiro state, actually in Brazil, indicating that the epidemiological importance of this vector in the transmission of diseases with great economic impact must not be ignored.

Attractivity of vinasse spraying to stable flies, Stomoxys calcitrans in a sugarcane area

ABSTRACT: Stable fly (Stomoxys calcitrans) outbreaks have been associated with sugarcane mills in Brazil. This study aimed to evaluate the attractivity of vinasse, a liquid byproduct of ethanol production applied to postharvest sugarcane fields, to adult S. calcitrans. Stable fly abundance was monitored for 21 days in August-September 2014, at a sugarcane mill in the municipality of Nova Alvorada do Sul, state of Mato Grosso do Sul, Brazil. Following mechanized harvesting of sugarcane and the consequent deposition of large amounts of plant residue (straw) in the field, the study area (13.5ha) was surrounded by 22 sticky cylinder (alsynite) traps. The area was fertigated with vinasse immediately after the traps were placed. Insects caught in the traps were collected daily at 7:00 a.m., 11:00 a.m. and 15:00 p.m. A total of 83,593 specimens of S. calcitrans were caught, of which 5,516 (6.6%) were attracted within the first five hours after vinasse application. The highest number of S. calcitrans was caught in the first week following vinasse application, with a peak on the 4th day (12,818 flies), which was influenced by adverse weather conditions on the previous days. This study confirms that fertigation of sugarcane fields by spraying vinasse after the harvest attracts S. calcitrans, thus quickly increasing their abundance over the days following application.

RESUMO: Surtos da mosca-dos-estábulos (Stomoxys calcitrans) têm sido associados a usinas sucroalcooleiras no Brasil. Este estudo objetivou avaliar a atratividade da aspersão de vinhaça, um subproduto líquido da produção de etanol aplicado após a colheita em áreas canavieiras, a adultos de S. calcitrans. A abundância desta mosca foi monitorada em uma usina de cana-de-açúcar por 21 dias, entre agosto e setembro de 2014, em Nova Alvorada do Sul/MS. Após a colheita mecanizada da cana e consequente deposição de grande quantidade de palha no campo, a área de estudo (13,5ha) foi cercada com 22 armadilhas adesivas cilíndricas (alsynite). Fertirrigação com vinhaça foi iniciada imediatamente após instalação das armadilhas. A coleta das moscas capturadas nas armadilhas foi realizada diariamente às 7:00 a.m., 11:00 a.m. e 15:00 p.m. Foram capturados 83.593 espécimes de S. calcitrans, sendo 5.516 (6,6%) atraídos nas primeiras cinco horas após a aplicação da vinhaça. Maior número de S. calcitrans foi capturado na primeira semana do estudo (53.937 moscas), com pico no 4º dia (12.818 moscas), influenciado pelas condições climáticas adversas ocorridas nos dias anteriores. O presente estudo confirma que o processo de fertirrigação de áreas canavieiras com aspersão de vinhaça após a colheita é atrativo à S. calcitrans, aumentando rapidamente sua abundância nos dias subsequentes à aplicação.

Infection and reinfection of Stomoxys calcitrans larvae (Diptera: Muscidae) by entomopathogenic nematodes in different times of exposure / Infecção e reinfecção de larvas de Stomoxys calcitrans (Diptera: Muscidae) por nematoides entomopatogênicos em diferentes tempos de exposição

Abstract Stomoxys calcitrans is a hematophagous dipteran. Several agents are used in biological control, including entomopathogenic nematodes (EPNs). Bioassay I involved an evaluation of the effect of Heterorhabditis bacteriophora HP88 and Heterorhabditis baujardi LPP7 on S. calcitrans larvae in different periods of exposure. Groups of 10 larvae were placed in Petri dishes and 200 EPNs/larva were added, which were divided into groups according to the exposure times of 2, 4, 6, 12, 24 and 48 hours. The purpose of Bioassay II was to evaluate the efficacy of the EPNs in infecting S. calcitrans larvae when they were isolated from stable fly larvae in Bioassay I. Groups of 10 larvae were placed in Petri dishes and 200 EPNs/larva were added. In bioassay I, H. bacteriophora caused mortality rates of 51.7, 83.3 and 91.7% in 12, 24 and 48 hours, respectively, while H. baujardi caused mortality rates of 9.3 (12h), 35 (24h) and 35% (48h). In Bioassay II, H. bacteriophora and H. baujardi resulted in mortality rates of 35% and 25%, respectively. It was concluded that the longest exposure times presented the highest larval mortality and that EPNs isolated from S. calcitrans are not efficient in controlling the larvae fly.

Resumo Stomoxys calcitrans é um díptero hematófago. Vários agentes são usados no controle biológico, incluindo nematoides entomopatogênicos (NEPs). O bioensaio I objetivou avaliar o efeito de Heterorhabditis bacteriophora HP88 e Heterorhabditis baujardi LPP7 sobre larvas de S. calcitrans em diferentes períodos de exposição. Grupos de 10 larvas foram colocados em placas de Petri, adicionou-se 200 NEPs/larva, e foram divididos de acordo com tempos de exposição de 2, 4, 6, 12, 24 e 48 horas. O objetivo do Bioensaio II foi avaliar a eficácia dos NEPs na infecção de larvas de S. calcitrans, quando esses foram isolados de larvas da mosca oriundos do Bioensaio I. Os grupos de 10 larvas foram colocados em placas de Petri e 200 NEPs/larva foram adicionados. No bioensaio I, H. bacteriophora causou taxas de mortalidade de 51,7%, 83,3% e 91,7% em 12, 24 e 48 horas, respectivamente, enquanto H. baujardi causou taxas de mortalidade de 9,3% (12h), 35% (24h) e 35% (48h). No Bioensaio II, H. bacteriophora e H. baujardi resultaram em taxas de mortalidade de 35% e 25%, respectivamente. Concluiu-se que os maiores tempos de exposição apresentaram as maiores mortalidades larvais e que NEPs isolados de S. calcitrans não são eficientes no controle das larvas mosca.

Attractivity of vinasse spraying to stable flies, Stomoxys calcitrans in a sugarcane area / C Stomoxys calcitrans (Diptera: Muscidae), em área canavieira

Stable fly (Stomoxys calcitrans) outbreaks have been associated with sugarcane mills in Brazil. This study aimed to evaluate the attractivity of vinasse, a liquid byproduct of ethanol production applied to postharvest sugarcane fields, to adult S. calcitrans. Stable fly abundance was monitored for 21 days in August-September 2014, at a sugarcane mill in the municipality of Nova Alvorada do Sul, state of Mato Grosso do Sul, Brazil. Following mechanized harvesting of sugarcane and the consequent deposition of large amounts of plant residue (straw) in the field, the study area (13.5ha) was surrounded by 22 sticky cylinder ("alsynite") traps. The area was fertigated with vinasse immediately after the traps were placed. Insects caught in the traps were collected daily at 7:00 a.m., 11:00 a.m. and 15:00 p.m. A total of 83,593 specimens of S. calcitrans were caught, of which 5,516 (6.6%) were attracted within the first five hours after vinasse application. The highest number of S. calcitrans was caught in the first week following vinasse application, with a peak on the 4th day (12,818 flies), which was influenced by adverse weather conditions on the previous days. This study confirms that fertigation of sugarcane fields by spraying vinasse after the harvest attracts S. calcitrans, thus quickly increasing their abundance over the days following application.(AU)

Surtos da mosca-dos-estábulos (Stomoxys calcitrans) têm sido associados a usinas sucroalcooleiras no Brasil. Este estudo objetivou avaliar a atratividade da aspersão de vinhaça, um subproduto líquido da produção de etanol aplicado após a colheita em áreas canavieiras, a adultos de S. calcitrans. A abundância desta mosca foi monitorada em uma usina de cana-de-açúcar por 21 dias, entre agosto e setembro de 2014, em Nova Alvorada do Sul/MS. Após a colheita mecanizada da cana e consequente deposição de grande quantidade de palha no campo, a área de estudo (13,5ha) foi cercada com 22 armadilhas adesivas cilíndricas ("alsynite"). Fertirrigação com vinhaça foi iniciada imediatamente após instalação das armadilhas. A coleta das moscas capturadas nas armadilhas foi realizada diariamente às 7:00 a.m., 11:00 a.m. e 15:00 p.m. Foram capturados 83.593 espécimes de S. calcitrans, sendo 5.516 (6,6%) atraídos nas primeiras cinco horas após a aplicação da vinhaça. Maior número de S. calcitrans foi capturado na primeira semana do estudo (53.937 moscas), com pico no 4º dia (12.818 moscas), influenciado pelas condições climáticas adversas ocorridas nos dias anteriores. O presente estudo confirma que o processo de fertirrigação de áreas canavieiras com aspersão de vinhaça após a colheita é atrativo à S. calcitrans, aumentando rapidamente sua abundância nos dias subsequentes à aplicação.(AU)

Infection and reinfection of Stomoxys calcitrans larvae (Diptera: Muscidae) by entomopathogenic nematodes in different times of exposure / Infecção e reinfecção de larvas de Stomoxys calcitrans (Diptera: Muscidae) por nematoides entomopatogênicos em diferentes tempos de exposição

Stomoxys calcitrans is a hematophagous dipteran. Several agents are used in biological control, including entomopathogenic nematodes (EPNs). Bioassay I involved an evaluation of the effect of Heterorhabditis bacteriophora HP88 and Heterorhabditis baujardi LPP7 on S. calcitrans larvae in different periods of exposure. Groups of 10 larvae were placed in Petri dishes and 200 EPNs/larva were added, which were divided into groups according to the exposure times of 2, 4, 6, 12, 24 and 48 hours. The purpose of Bioassay II was to evaluate the efficacy of the EPNs in infecting S. calcitrans larvae when they were isolated from stable fly larvae in Bioassay I. Groups of 10 larvae were placed in Petri dishes and 200 EPNs/larva were added. In bioassay I, H. bacteriophora caused mortality rates of 51.7, 83.3 and 91.7% in 12, 24 and 48 hours, respectively, while H. baujardi caused mortality rates of 9.3 (12h), 35 (24h) and 35% (48h). In Bioassay II, H. bacteriophora and H. baujardi resulted in mortality rates of 35% and 25%, respectively. It was concluded that the longest exposure times presented the highest larval mortality and that EPNs isolated from S. calcitrans are not efficient in controlling the larvae fly.(AU)

Stomoxys calcitrans é um díptero hematófago. Vários agentes são usados no controle biológico, incluindo nematoides entomopatogênicos (NEPs). O bioensaio I objetivou avaliar o efeito de Heterorhabditis bacteriophora HP88 e Heterorhabditis baujardi LPP7 sobre larvas de S. calcitrans em diferentes períodos de exposição. Grupos de 10 larvas foram colocados em placas de Petri, adicionou-se 200 NEPs/larva, e foram divididos de acordo com tempos de exposição de 2, 4, 6, 12, 24 e 48 horas. O objetivo do Bioensaio II foi avaliar a eficácia dos NEPs na infecção de larvas de S. calcitrans, quando esses foram isolados de larvas da mosca oriundos do Bioensaio I. Os grupos de 10 larvas foram colocados em placas de Petri e 200 NEPs/larva foram adicionados. No bioensaio I, H. bacteriophora causou taxas de mortalidade de 51,7%, 83,3% e 91,7% em 12, 24 e 48 horas, respectivamente, enquanto H. baujardi causou taxas de mortalidade de 9,3% (12h), 35% (24h) e 35% (48h). No Bioensaio II, H. bacteriophora e H. baujardi resultaram em taxas de mortalidade de 35% e 25%, respectivamente. Concluiu-se que os maiores tempos de exposição apresentaram as maiores mortalidades larvais e que NEPs isolados de S. calcitrans não são eficientes no controle das larvas mosca.(AU)

Attractivity of vinasse spraying to stable flies, Stomoxys calcitrans in a sugarcane area / C Stomoxys calcitrans (Diptera: Muscidae), em área canavieira

Stable fly (Stomoxys calcitrans) outbreaks have been associated with sugarcane mills in Brazil. This study aimed to evaluate the attractivity of vinasse, a liquid byproduct of ethanol production applied to postharvest sugarcane fields, to adult S. calcitrans. Stable fly abundance was monitored for 21 days in August-September 2014, at a sugarcane mill in the municipality of Nova Alvorada do Sul, state of Mato Grosso do Sul, Brazil. Following mechanized harvesting of sugarcane and the consequent deposition of large amounts of plant residue (straw) in the field, the study area (13.5ha) was surrounded by 22 sticky cylinder ("alsynite") traps. The area was fertigated with vinasse immediately after the traps were placed. Insects caught in the traps were collected daily at 7:00 a.m., 11:00 a.m. and 15:00 p.m. A total of 83,593 specimens of S. calcitrans were caught, of which 5,516 (6.6%) were attracted within the first five hours after vinasse application. The highest number of S. calcitrans was caught in the first week following vinasse application, with a peak on the 4th day (12,818 flies), which was influenced by adverse weather conditions on the previous days. This study confirms that fertigation of sugarcane fields by spraying vinasse after the harvest attracts S. calcitrans, thus quickly increasing their abundance over the days following application.(AU)

Surtos da mosca-dos-estábulos (Stomoxys calcitrans) têm sido associados a usinas sucroalcooleiras no Brasil. Este estudo objetivou avaliar a atratividade da aspersão de vinhaça, um subproduto líquido da produção de etanol aplicado após a colheita em áreas canavieiras, a adultos de S. calcitrans. A abundância desta mosca foi monitorada em uma usina de cana-de-açúcar por 21 dias, entre agosto e setembro de 2014, em Nova Alvorada do Sul/MS. Após a colheita mecanizada da cana e consequente deposição de grande quantidade de palha no campo, a área de estudo (13,5ha) foi cercada com 22 armadilhas adesivas cilíndricas ("alsynite"). Fertirrigação com vinhaça foi iniciada imediatamente após instalação das armadilhas. A coleta das moscas capturadas nas armadilhas foi realizada diariamente às 7:00 a.m., 11:00 a.m. e 15:00 p.m. Foram capturados 83.593 espécimes de S. calcitrans, sendo 5.516 (6,6%) atraídos nas primeiras cinco horas após a aplicação da vinhaça. Maior número de S. calcitrans foi capturado na primeira semana do estudo (53.937 moscas), com pico no 4º dia (12.818 moscas), influenciado pelas condições climáticas adversas ocorridas nos dias anteriores. O presente estudo confirma que o processo de fertirrigação de áreas canavieiras com aspersão de vinhaça após a colheita é atrativo à S. calcitrans, aumentando rapidamente sua abundância nos dias subsequentes à aplicação.(AU)

Establishment and quantitative measure of Stomoxys calcitrans (Linnaeus, 1758) colony production in Rio de Janeiro, Brazil.

Functional insect vector colonies are essential for the study of their biology, evolution, behavior as well as control strategies of these organisms that transmit pathogens of medical and veterinary importance. In addition to the establishment, improvement and adaptation of pre-established protocols in the maintenance of a colony is important, as it may result in higher production of insects. Stomoxys calcitrans or stable fly causes important economic losses in production of beef and milk, besides mechanically transmitting a great variety of pathogens to cattle. In order to generate flies under laboratory conditions, a colony of S. calcitrans was established at our laboratory considering local environment geographical characteristics. Adults collected in UFRRJ campus were kept in the laboratory and maintained with daily feeding on anticoagulant-containing bovine blood offered in disposable sanitary napkins. Immature forms were maintained on diets rich in organic matter. The colony temperature and relative humidity were daily monitored as well as the development of insects. The monthly measured biological parameters included the development of eggs into pupae, of pupae into F1 adults and the comparison of the number of flies collected in the field with the number of F1 flies emerged in the laboratory. The data presented here was obtained from March to November 2019. In that period, the average per month of flies collected in the field was 604.2, of generated eggs was 2694.3, of generated pupae was 349.5 and of generated F1 flies in the laboratory was 205.9. Fluctuations of abiotic and biological factors contributed to an uneven production of flies in the colony throughout the months. This is the first report with numerical quantification of S. calcitrans flies production in a laboratory in Brazil. Some methodologies used in the colony shall be reevaluated and modified with the goal of obtaining higher numbers of flies generated in the laboratory over time.

Adulticidal Activity of Melaleuca alternifolia (Myrtales: Myrtaceae) Essential Oil With High 1,8-Cineole Content Against Stable Flies (Diptera: Muscidae).

The stable fly, Stomoxys calcitrans (Linnaeus 1758), is a hematophagous fly responsible for causing loss of performance in horses, causing losses in cattle productivity, and impacting the animals' health through the spread of pathogenic microorganisms. The objective of this work was to investigate the insecticidal activity of essential oil obtained from Melaleuca alternifolia (Cheel), presenting high 1,8-cineole content, against S. calcitrans adults. Insecticidal activity was determined using surface application methods and exposure to oil impregnated paper. It was observed that treatments at 25 and 50 µg/cm2 (P < 0.05) present fumigant activity through exposure to the impregnated paper, and in the first 15 min of exposure, the mortality rates obtained for these treatments were, respectively (96.6 ± 3.3% and 100%), equivalent to the positive control. Using the superficial application method, the only treatment concentration presenting adulticidal action was 5% (w/v) (P < 0.05). Respective toxicities LC50 (%, w/v) and LC80 for the impregnated paper method were 1.06 ± 0.02 and 1.47 ± 0.17; for the superficial application method, they were 3.82 ± 0.65 and 5.53 ± 0.74. As demonstrated, M. alternifolia essential oil presents adulticidal potential against S. calcitrans.

Resistance of the stable fly, Stomoxys calcitrans (Diptera: Muscidae), to cypermethrin in outbreak areas in Midwestern Brazil / Resistência da mosca-dos-estábulos, Stomoxys calcitrans (Diptera: Muscidae), à cipermetrina em áreas de surtos no Centro-Oeste brasileiro

Abstract Population explosions of the stable fly (Stomoxys calcitrans) have become a serious concern for livestock producers near sugarcane mills in some regions of Brazil due to the insect's massive reproduction on sugarcane byproducts and waste. Despite the limited efficiency of insecticides for controlling stable fly outbreaks, producers still rely on chemical control to mitigate the alarming infestations in affected areas. This study evaluated the susceptibility of S. calcitrans populations to cypermethrin in the state of Mato Grosso do Sul, Brazil. Stable flies were tested from three field populations and two colonies, established from flies previously collected at sugarcane mills. Wild flies were collected with Nzi traps in areas of sugarcane plantations. Both wild and colonized flies were exposed to eleven concentrations of cypermethrin in impregnated filter paper bioassays. All the populations proved to be resistant to cypermethrin, with resistance factors among field populations ranging from 6.8 to 38.6. The intensive use of insecticides has led to the development of pyrethroid resistance in stable fly populations in the proximities of sugarcane mills in the state of Mato Grosso do Sul.

Resumo Explosões populacionais da mosca-dos-estábulos (Stomoxys calcitrans) tornaram-se uma séria preocupação para a pecuária próxima a usinas de cana-de-açúcar em algumas regiões do Brasil, devido à massiva reprodução da mosca em resíduos e subprodutos do processamento da cana. Apesar da limitada eficiência dos inseticidas no controle dos surtos desta mosca, produtores dependem do controle químico para mitigar as alarmantes infestações nas áreas afetadas. Este estudo objetivou avaliar a suscetibilidade de populações de S. calcitrans a piretroides no estado de Mato Grosso do Sul, Brasil. Moscas-dos-estábulos de três populações de campo e duas colônias, estabelecidas a partir de moscas previamente coletadas em usinas de cana-de-açúcar, foram testadas. Moscas selvagens foram coletadas com armadilhas Nzi em áreas de cana. Moscas coletadas foram expostas a onze concentrações de cipermetrina em bioensaios com papel de filtro impregnado. Todas as populações testadas mostraram-se resistentes ao piretroide, com fatores de resistência variando de 6,8 a 38,6 nas populações de campo. O uso intensivo de inseticidas tem levado ao desenvolvimento de resistência da mosca-dos-estábulos a piretroides em populações próximas a usinas de cana-de-açúcar no estado do Mato Grosso do Sul.

Control of Haematobia irritans and Stomoxys calcitrans with Metarhizium anisopliae in naturally infested cattle / Control de Haematobia irritans y Stomoxys calcitrans con Metarhizium anisopliae en ganado naturalmente infestado

ABSTRACT Objective. Assess the efficacy of Metarhizium anisopliae strain 135 (Ma135) applied by aspersion to reduce simultaneous infestation of Haematobia irritans and Stomoxys calcitrans in naturally infested cattle maintained in a production system which combine grazing and confinement in pens. Materials and methods. Was applied an aqueous formulation of Ma135 (1x108 conidia/ml) on a nine cows group in six occasions with seven days interval, while the control group received the same formulation without conidial content. The infestation index was estimated daily for each fly independently; the effectiveness of the formulation was calculated using the Abbott's formula. Results. The Ma135 formulation had a reduction in the fly population of 58% for H. irritans and 69% for S. calcitrans, taking into consideration the six study weeks, in addition to causing no negative effects on animal health. An Infestation index reduction was observed from the first week post-treatment (p<0.05) and maintained this trend throughout the study. Conclusions. The present study has demonstrated the potential of strain Ma135 to reduce the simultaneous infestation of both hematophagous flies in cattle under conditions of natural infestation.

RESUMEN Objetivo. Evaluar la eficacia de la cepa 135 de Metarhizium anisopliae (Ma135) aplicada por aspersión para reducir la infestación simultánea de Haematobia irritans y Stomoxys calcitrans en ganado naturalmente infestado mantenido en un sistema de producción de leche que combina el pastoreo y el confinamiento en corrales. Materiales y métodos. Se aplicó una formulación acuosa de Ma135 (1x108 conidios/ml) a un grupo de nueve vacas en seis ocasiones con un intervalo de siete días, mientras que el grupo control recibió una solución compuesta por agua, Monooleato de polioxietileno sorbitán en solución 0.01% y un adyuvante agrícola al 0.1%. Se estimó diariamente el índice de infestación para cada mosca de forma independiente; la efectividad de la formulación se calculó usando la fórmula de Abbott. Resultados. La formulación de Ma135 tuvo una eficacia en el control de la infestación de 58% para el caso de H. irritans y de 69% para S. calcitrans, tomando en cuenta las seis semanas de estudio, además de que no causó ningún efecto negativo en la salud de los animales. La reducción del índice de infestación se observó desde la primera semana post-tratamiento (p<0.05) y mantuvo esta tendencia durante todo el estudio. Conclusiones. El presente estudio ha demostrado el potencial de la cepa Ma135 para reducir la infestación simultánea de ambas moscas hematófagas en el ganado bajo condiciones de infestación natural.

Resistance of the stable fly, Stomoxys calcitrans (Diptera: Muscidae), to cypermethrin in outbreak areas in Midwestern Brazil / Resistência da mosca-dos-estábulos, Stomoxys calcitrans (Diptera: Muscidae), à cipermetrina em áreas de surtos no Centro-Oeste brasileiro

Population explosions of the stable fly (Stomoxys calcitrans) have become a serious concern for livestock producers near sugarcane mills in some regions of Brazil due to the insects massive reproduction on sugarcane byproducts and waste. Despite the limited efficiency of insecticides for controlling stable fly outbreaks, producers still rely on chemical control to mitigate the alarming infestations in affected areas. This study evaluated the susceptibility of S. calcitrans populations to cypermethrin in the state of Mato Grosso do Sul, Brazil. Stable flies were tested from three field populations and two colonies, established from flies previously collected at sugarcane mills. Wild flies were collected with Nzi traps in areas of sugarcane plantations. Both wild and colonized flies were exposed to eleven concentrations of cypermethrin in impregnated filter paper bioassays. All the populations proved to be resistant to cypermethrin, with resistance factors among field populations ranging from 6.8 to 38.6. The intensive use of insecticides has led to the development of pyrethroid resistance in stable fly populations in the proximities of sugarcane mills in the state of Mato Grosso do Sul.(AU)

Explosões populacionais da mosca-dos-estábulos (Stomoxys calcitrans) tornaram-se uma séria preocupação para a pecuária próxima a usinas de cana-de-açúcar em algumas regiões do Brasil, devido à massiva reprodução da mosca em resíduos e subprodutos do processamento da cana. Apesar da limitada eficiência dos inseticidas no controle dos surtos desta mosca, produtores dependem do controle químico para mitigar as alarmantes infestações nas áreas afetadas. Este estudo objetivou avaliar a suscetibilidade de populações de S. calcitrans a piretroides no estado de Mato Grosso do Sul, Brasil. Moscas-dos-estábulos de três populações de campo e duas colônias, estabelecidas a partir de moscas previamente coletadas em usinas de cana-de-açúcar, foram testadas. Moscas selvagens foram coletadas com armadilhas Nzi em áreas de cana. Moscas coletadas foram expostas a onze concentrações de cipermetrina em bioensaios com papel de filtro impregnado. Todas as populações testadas mostraram-se resistentes ao piretroide, com fatores de resistência variando de 6,8 a 38,6 nas populações de campo. O uso intensivo de inseticidas tem levado ao desenvolvimento de resistência da mosca-dos-estábulos a piretroides em populações próximas a usinas de cana-de-açúcar no estado do Mato Grosso do Sul.(AU)

Potential of entomopathogenic nematodes of the genus Heterorhabditis for the control of Stomoxys calcitrans (Diptera: Muscidae) / Potencial de nematoides entomopatogênicos do gênero Heterorhabditis para o controle de Stomoxys calcitrans (Diptera: Muscidae)

Abstract The present study verified the pathogenic potential of entomopathogenic nematodes (EPNs) of the genus Heterorhabditis (Heterorhabditis bacteriophora, isolate HP88 and Heterorhabditis baujardi isolate LPP7) to immature stages of Stomoxys calcitrans in the laboratory. All EPN concentrations of the H. bacteriophora HP88 strain caused mean larval mortality greater than 90% after four days. Higher concentrations of the H. baujardi LPP7 isolate (≥50 EPNs/larva) eliminated more than 70% of larvae after six days with the concentration 200 EPNs/larva reaching mortality levels of 93.3%. The larval mortality at all concentrations of EPNs (25, 50, 100, 150, and 200 EPNs/larva) for both strains was significant (p<0.05) when compared to the respective control groups. Concentrations of H. bacteriophora HP88 yielded an LC50 of 0.36 EPN/larva and LC90 of 29.1; while H. baujardi LPP7 yielded an LC50 of 39.85 and LC90 of 239.18. H. bacteriophora HP88 provided greater inhibition of the emergence of adults when compared to the response obtained with H. baujardi LPP7. EPNs did not cause considerable mortality when applied directly to pupae. The set of observed results suggests that the EPNs of the genus Heterorhabditis, isolates HP88 and LPP7, are a promising alternative in the control of the stable fly.

Resumo O presente estudo verificou o potencial patogênico de nematoides (NEP) do gênero Heterorhabditis (H. bacteriophora - isolado HP88 e H. baujardi - isolado LPP7) para estágios imaturos de Stomoxys calcitrans em laboratório. Todas as concentrações de NEPs da cepa HP88 causaram mortalidade larval média maior que 90% após quatro dias. Concentrações mais elevadas da cepa LPP7 (≥50 NEPs/larva) eliminaram mais de 70% após seis dias, com a concentração 200 NEPs/larva atingindo níveis de 93,3%. A mortalidade larval em todas as concentrações de NEPs (25, 50, 100, 150, and 200 NEPs/larva) para ambas as cepas foi significativa (p<0,05), quando comparadas aos respectivos grupos controle. Concentrações de H. bacteriophora HP88 obtiveram LC50 de 0,36 NEP/larva e LC90 de 29,1; enquanto H. baujardi LPP7 obteve LC50 de 39,85 e LC90 de 239,18. O isolado H. bacteriophora HP88 propiciou maior inibição da emergência de adultos, quando comparado à resposta obtida com H. baujardi LPP7. NEPs não provocaram mortalidade considerável quando aplicados diretamente sobre pupas. O conjunto de resultados observados indica os nematoides entomopatogênicos do gênero Heterorhabditis (estirpes HP88 e LPP7) como uma alternativa promissora no controle da mosca dos estábulos.

Potential of entomopathogenic nematodes of the genus Heterorhabditis for the control of Stomoxys calcitrans (Diptera: Muscidae) / Potencial de nematoides entomopatogênicos do gênero Heterorhabditis para o controle de Stomoxys calcitrans (Diptera: Muscidae)

The present study verified the pathogenic potential of entomopathogenic nematodes (EPNs) of the genus Heterorhabditis (Heterorhabditis bacteriophora, isolate HP88 and Heterorhabditis baujardi isolate LPP7) to immature stages of Stomoxys calcitrans in the laboratory. All EPN concentrations of the H. bacteriophora HP88 strain caused mean larval mortality greater than 90% after four days. Higher concentrations of the H. baujardi LPP7 isolate (≥50 EPNs/larva) eliminated more than 70% of larvae after six days with the concentration 200 EPNs/larva reaching mortality levels of 93.3%. The larval mortality at all concentrations of EPNs (25, 50, 100, 150, and 200 EPNs/larva) for both strains was significant (p<0.05) when compared to the respective control groups. Concentrations of H. bacteriophora HP88 yielded an LC50 of 0.36 EPN/larva and LC90 of 29.1; while H. baujardi LPP7 yielded an LC50 of 39.85 and LC90 of 239.18. H. bacteriophora HP88 provided greater inhibition of the emergence of adults when compared to the response obtained with H. baujardi LPP7. EPNs did not cause considerable mortality when applied directly to pupae. The set of observed results suggests that the EPNs of the genus Heterorhabditis, isolates HP88 and LPP7, are a promising alternative in the control of the stable fly.(AU)

O presente estudo verificou o potencial patogênico de nematoides (NEP) do gênero Heterorhabditis (H. bacteriophora- isolado HP88 e H. baujardi - isolado LPP7) para estágios imaturos de Stomoxys calcitrans em laboratório. Todas as concentrações de NEPs da cepa HP88 causaram mortalidade larval média maior que 90% após quatro dias. Concentrações mais elevadas da cepa LPP7 (≥50 NEPs/larva) eliminaram mais de 70% após seis dias, com a concentração 200 NEPs/larva atingindo níveis de 93,3%. A mortalidade larval em todas as concentrações de NEPs (25, 50, 100, 150, and 200 NEPs/larva) para ambas as cepas foi significativa (p<0,05), quando comparadas aos respectivos grupos controle. Concentrações de H. bacteriophora HP88 obtiveram LC50 de 0,36 NEP/larva e LC90 de 29,1; enquanto H. baujardi LPP7 obteve LC50 de 39,85 e LC90 de 239,18. O isolado H. bacteriophora HP88 propiciou maior inibição da emergência de adultos, quando comparado à resposta obtida com H. baujardi LPP7. NEPs não provocaram mortalidade considerável quando aplicados diretamente sobre pupas. O conjunto de resultados observados indica os nematoides entomopatogênicos do gênero Heterorhabditis (estirpes HP88 e LPP7) como uma alternativa promissora no controle da mosca dos estábulos.(AU)

Identification of stable fly attractant compounds in vinasse, a byproduct of sugarcane-ethanol distillation.

The stable fly, Stomoxys calcitrans (Diptera: Muscidae), is a worldwide pest of livestock. Recent outbreaks of stable flies in sugarcane fields in Brazil have become a serious problem for livestock producers. Larvae and pupae found inside sugarcane stems after harvesting may indicate that stable flies use these stems as potential oviposition or larval development sites. Field observations suggest that outbreaks of stable flies are associated with the vinasse and filter cake derived from biomass distillation in sugarcane ethanol production that are used as fertilizers in sugarcane fields. Adult stable flies are attracted to vinasse, which appears to present an ideal larval development site. The primary goal of the present study is to demonstrate the role of vinasse in influencing the sensory physiological and behavioural responses of stable flies, and to identify its associated volatile attractant compounds. Both laboratory and field studies showed that vinasse is extremely attractive to adult stable flies. Chemical analyses of volatiles collected revealed a wide range of carboxylic acids, alcohols, phenols and aldehydes as potential attractant compounds. These newly identified attractants could be used to develop a tool for the attractant-baited mass trapping of stable flies in order to reduce infestations.

Checklist de Muscidae (Insecta, Diptera) do Estado de Mato Grosso do Sul / Checklist of Muscidae (Insecta, Diptera) from Mato Grosso do Sul state

A checklist of Muscidae species in the state of Mato Grosso do Sul, Brazil is provided. There are 65 species recorded, which represent 17% of all Brazilian species of Muscidae known.(AU)

É apresentada uma lista de espécies do estado de Mato Grosso do Sul. Existem 65 espécies registradas, representado 17% de todas as espécies de Muscidae conhecidas no Brasil.(AU)