Insecticide Sensitivity of Native Chloride and Sodium Channels in a Mosquito Cell Line

The aim of this study was to investigate the utility of cultured Anopheles gambiae Sua1B cells for insecticide screening applications without genetic engineering or other treatments. Sua1B cells were exposed to the known insecticidal compounds lindane and DIDS, which inhibited cell growth at micromolar concentrations. In patch clamp studies, DIDS produced partial inhibition (69%) of chloride current amplitudes, and an IC50 of 5.1 μM was determined for Sua1B cells. A sub-set of chloride currents showed no response to DIDS; however, inhibition (64%) of these currents was achieved using a low chloride saline solution, confirming their identity as chloride channels. In contrast, lindane increased chloride current amplitude (EC50 = 116 nM), which was reversed when cells were bathed in calcium-free extracellular solution. Voltage-sensitive chloride channels were also inhibited by the presence of fenvalerate, a type 2 pyrethroid, but not significantly blocked by type 1 allethrin, an effect not previously shown in insects. Although no evidence of fast inward currents typical of sodium channels was observed, studies with fenvalerate in combination with veratridine, a sodium channel activator, revealed complete inhibition of cell growth that was best fit by a two-site binding model. The high potency effect was completely inhibited in the presence of tetrodotoxin, a specific sodium channel blocker, suggesting the presence of some type of sodium channel. Thus, Sua1B cells express native insect ion channels with potential utility for insecticide screening

A survey of the eutrophication state of an urbanized tropical estuary, the case of the Great Vitória Estuarine System, Brazil.

Although, estuarine ecosystems have an ecological and economical importance as they have a high  biological productivity and occur within a unique and dynamic environment, they have been subjected to anthropogenic alterations. The Great Vitória Estuarine System (GVES), Espírito Santo State, Brazil, is not an exception, as urbanization is growing around it with a high quantity of sewage been added to the system. In order to evaluate the eutrophication state as well as the meiofauna response to it, several parameters in both sediments and in the water column were assessed. Orthophosphate, nitrite, nitrate, ammonia, chla and Fecal Coliform counts (FC) ranged from 0.2 to 3.2 μM, 0.25 to 1.14 μM, 1.83 to 0.19μM, 4.19 to 49.23 μM, 0.61 to 6.72 μg/L and 14 to 5.0x104 MPN/100 mL of water, respectively. These results showed that the GVES is under an eutrophication process and that the Passagem Channel experienced the largest impacts. Sewage plays an important role in this eutrophication process as indicated by PCA and correlations tests. The density of meiofauna showed similar values to those found in environments with similar levels of anthropogenic stress. This multi-approach evaluation revealed several aspects of the impacted estuary and could be used as an important tool to manage better the estuary.Although, estuarine ecosystems have an ecological and economical importance as they have a high  biological productivity and occur within a unique and dynamic environment, they have been subjected to anthropogenic alterations. The Great Vitória Estuarine System (GVES), Espírito Santo State, Brazil, is not an exception, as urbanization is growing around it with a high quantity of sewage been added to the system. In order to evaluate the eutrophication state as well as the meiofauna response to it, several parameters in both sediments and in the water column were assessed. Orthophosphate, nitrite, nitrate, ammonia, chla and Fecal Coliform counts (FC) ranged from 0.2 to 3.2 μM, 0.25 to 1.14 μM, 1.83 to 0.19μM, 4.19 to 49.23 μM, 0.61 to 6.72 μg/L and 14 to 5.0x104 MPN/100 mL of water, respectively. These results showed that the GVES is under an eutrophication process and that the Passagem Channel experienced the largest impacts. Sewage plays an important role in this eutrophication process as indicated by PCA and correlations tests. The density of meiofauna showed similar values to those found in environments with similar levels of anthropogenic stress. This multi-approach evaluation revealed several aspects of the impacted estuary and could be used as an important tool to manage better the estuary

Orange jasmine as a trap crop to control Diaphorina citri

[EN] Novel, suitable and sustainable alternative control tactics that have the potential to reduce migration of Diaphorina citri into commercial citrus orchards are essential to improve management of huanglongbing (HLB). In this study, the effect of orange jasmine (Murraya paniculata) as a border trap crop on psyllid settlement and dispersal was assessed in citrus orchards. Furthermore, volatile emission profiles and relative attractiveness of both orange jasmine and sweet orange (Citrus¿×¿aurantium L., syn. Citrus sinensis (L.) Osbeck) nursery flushes to D. citri were investigated. In newly established citrus orchards, the trap crop reduced the capture of psyllids in yellow sticky traps and the number of psyllids that settled on citrus trees compared to fallow mowed grass fields by 40% and 83%, respectively. Psyllids were attracted and killed by thiamethoxam-treated orange jasmine suggesting that the trap crop could act as a `sink¿ for D. citri. Additionally, the presence of the trap crop reduced HLB incidence by 43%. Olfactometer experiments showed that orange jasmine plays an attractive role on psyllid behavior and that this attractiveness may be associated with differences in the volatile profiles emitted by orange jasmine in comparison with sweet orange. Results indicated that insecticide-treated M. paniculata may act as a trap crop to attract and kill D. citri before they settled on the edges of citrus orchards, which significantly contributes to the reduction of HLB primary spread.This work was supported by Fund for Citrus Protection (Fundecitrus) and by Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) (Proc. 2015/07011-3). We thank Moacir Celio Vizone, Felipe Marinho Martini and Joao Pedro Ancoma Lopes for technical support with experiments. Furthermore, we thank Cambuhy Agricola Ltda. and University of Araraquara (Uniara) for providing the areas in which the field experiments were performed. 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Susceptibility of Tribolium castaneum life stages exposed to elevated temperatures during heat treatments of a pilot flour mill: influence of sanitation, temperatures attained among mills floors, and costs

The influence of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), was investigated in a pilot flour mill subjected to three, 24-h heat treatments by using forced-air gas heaters fueled by propane. Two sanitation levels, dusting of wheat flour and 2-cm-deep flour, were created in 25 plastic bioassay boxes, each holding eggs, young larvae, old larvae, pupae, and adults of T. castaneum plus two temperature sensors. Data loggers (48) were placed on the five mill floors to record air temperatures. The time required to reach 50°C, time above 50°C, and the maximum temperature among mill floors and in bioassay boxes were measured. The maximum temperature in bioassay boxes and in the mill was lower on the first floor than on other floors. This trend was apparent in time required to reach 50°C and time above 50°C, especially in compartments with 2-cm-deep flour. The mean ± SE mortality of T. castaneum life stages on the first floor was 55.5 ± 12.9–98.6 ± 0.8%; it was 93.2 ± 6.7–100 ± 0.0% on other floors. Adults were the least susceptible stage. Mortality of T. castaneum stages in compartments with 2-cm-deep flour was generally lower than those with flour dust. Costs for the three heat treatments ranged from US$27,438 to$28,838. An effective heat treatment can be conducted within 24 h, provided temperatures on mill floors reach 50°C in 8–12 h and are held above 50°C for at least 10–14 h, with maximum temperatures held between 50 and 60°C

Biochemical and Hematologic Manifestations of Gastric Intrinsic Factor (GIF) Deficiency: A Treatable Cause of B12 Deficiency in the Old Order Mennonite Population of Southwestern Ontario

Intrinsic factor deficiency (OMIM #261000, IFD) is a rare inherited disorder of vitamin B12 metabolism due to mutations in the gastric intrinsic factor (GIF) gene. We report three individuals from an Old Order Mennonite community who presented with B12 deficiency. Two cases are siblings born to consanguineous parents and the third case is not known to be closely related. The older male sib presented at 4 years with gastrointestinal symptoms, listlessness, and pallor. He had pancytopenia with megaloblastic anemia. Serum B12 was 61 (198–615 pmol/L). Methylmalonic aciduria was present. C3 was elevated on acylcarnitine profile. Homocysteine was high at 16.7 (5.0–12.0 umol/L). His asymptomatic female sibling was also found to have B12 deficiency. Genetic testing for methylmalonic aciduria (MMAA), transcobalamin deficiency (TCN2), and Imerslund-Gräsbeck syndrome (AMN) showed no mutation in both siblings. The third patient, a 34-year-old woman, had presented in infancy with a diagnosis of pernicious anemia. Mutation analysis of GIF revealed compound heterozygosity for a c.79+1G\u3eA substitution and a c.973delG deletion in all three individuals. Oral or parenteral vitamin B12 has led to complete recovery of clinical parameters and vitamin B12 levels. Newborn screening samples on the siblings revealed normal methylcitrate, C3, and C3/C2 ratios thus indicating no disruption of propionic or methylmalonic acid metabolism. A high index of suspicion should be maintained if children present with megaloblastic anemia since GIF deficiency is a treatable disorder and newborn screening may not be able to detect this condition