This study further confirms the association of the IVS30+G>T mutation of the Tg gene with hypothyroidism. Computer analysis predicts that the A2215D mutation, first reported here, should cause structural instability of Tg but when present as a compound heterozygous mutation with IVS30+G>T/A its effect is unclear but is likely to be influenced by iodine intake.

BackgroundEntomopathogenic fungi are potential candidates for use in integrated vector management and many isolates are compatible with synthetic and natural insecticides. Neem oil was tested separately and in combination with the entomopathogenic fungus Metarhizium anisopliae against larvae of the dengue vector Aedes aegypti. Our aim was to increase the effectiveness of the fungus for the control of larval mosquito populations.MethodsCommercially available neem oil was used at concentrations ranging from 0.0001 to 1 %. Larval survival rates were monitored over a 7 day period following exposure to neem. The virulence of the fungus M. anisopliae was confirmed using five conidial concentrations (1 × 105 to 1 × 109 conidia mL−1) and survival monitored over 7 days. Two concentrations of fungal conidia were then tested together with neem (0.001 %). Survival curve comparisons were carried out using the Log-rank test and end-point survival rates were compared using one-way ANOVA.Results1 % neem was toxic to A. aegypti larvae reducing survival to 18 % with S50 of 2 days. Neem had no effect on conidial germination or fungal vegetative growth in vitro. Larval survival rates were reduced to 24 % (S50 = 3 days) when using 1 × 109 conidia mL−1. Using 1 × 108 conidia mL−1, 30 % survival (S50 = 3 days) was observed. We tested a “sub-lethal” neem concentration (0.001 %) together with these concentrations of conidia. For combinations of neem + fungus, the survival rates were significantly lower than the survival rates seen for fungus alone or for neem alone. Using a combination of 1 × 107 conidia mL−1 + neem (0.001 %), the survival rates were 36 %, whereas exposure to the fungus alone resulted in 74 % survival and exposure to neem alone resulted in 78 % survival. When using 1 × 108 conidia mL−1, the survival curves were modified, with a combination of the fungus + neem resulting in 12 % survival, whilst the fungus alone at this concentration also significantly reduced survival rates (28 %).ConclusionsThe use of adjuvants is an important strategy for maintaining/increasing fungal virulence and/or shelf-life. The addition of neem to conidial suspensions improved virulence, significantly reducing larval survival times and percentages.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-1280-9) contains supplementary material, which is available to authorized users.

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The mosquito Aedes aegypti is the most notorious vector of illness-causing viruses. The use of entomopathogenic fungi as bioinsecticides is a promising alternative for the development of novel mosquito control strategies. We investigate whether differences in immune responses could be responsible for modifications in survival rates of insects following different feeding regimes. Sucrose and blood-fed adult A. aegypti females were sprayed with M. anisopliae 1 × 106 conidia mL−1, and after 48 h, the midgut and fat body were dissected. We used RT-qPCR to monitor the expression of Cactus and REL1 (Toll pathway), IMD, REL2, and Caspar (IMD pathway), STAT and PIAS (JAK-STAT pathway), as well as the expression of antimicrobial peptides (Defensin A, Attacin and Cecropin G). REL1 and REL2 expression in both the midgut and fat body were higher in blood-fed fungus-challenged A. aegypti than in sucrose-fed counterparts. Interestingly, infection of sucrose-fed insects induced Cactus expression in the fat body, a negative regulator of the Toll pathway. The IMD gene was upregulated in the fat body in response to fungal infection after a blood meal. Additionally, we observed the induction of antimicrobial peptides in the blood-fed fungus-challenged insects. This study suggests that blood-fed A. aegypti are less susceptible to fungal infection due to the rapid induction of Toll and IMD immune pathways.

Recently, advances have been made in the use of entomopathogenic fungi for the control of not only crop pests but also disease vectoring insects. New approaches, for example, combining control strategies such as the application of conventional insecticides and biological control agents together, are highly promising, especially as many insecticides are compatible with entomopathogenic fungi. In this review, we discuss some of the new approaches being developed for vector control. Governmental authorities must implement rational integrated vector management programs for reducing deaths and suffering caused by insect-vectored diseases. Although there are now many alternatives to conventional chemical control methods, the use of pesticides is still the mainstay of mosquito control measures. However, mosquitoes are rapidly developing resistance to the chemical insecticides in use at the moment. The deployment of entomopathogenic fungi for the control of all stages of the mosquito life cycle is an alternative strategy currently being investigated by many different research groups. It has been shown that entomopathogenic fungi can efficiently kill mosquito larvae and adults in laboratory and field conditions. However, an important aspect of this new approach is how to apply these biological control agents economically and efficiently. By using combinations of control agents and novel application systems, it may be possible to significantly reduce mosquito populations to levels at which epidemics of malaria, dengue fever, zika virus, and chikungunya do not occur.

the soil Nitrogen Availability predictor (sNAp) model predicts daily and annual rates of net N mineralization (NNm) based on daily weather measurements, daily predictions of soil water and soil temperature, and on temperature and moisture modifiers obtained during aerobic incubation (basal rate). the model was based on in situ measurements of NNm in Australian soils under temperate climate. the purpose of this study was to assess this model for use in tropical soils under eucalyptus plantations in São Paulo State, Brazil. Based on field incubations for one month in three, NNm rates were measured at 11 sites (0-20 cm layer) for 21 months. the basal rate was determined in in situ incubations during moist and warm periods (January to march). Annual rates of 150-350 kg ha -1 yr -1 NNm predicted by the sNAp model were reasonably accurate (r 2 = 0.84). in other periods, at lower moisture and temperature, NNm rates were overestimated. therefore, if used carefully, the model can provide adequate predictions of annual NNm and may be useful in practical applications. for NNm predictions for shorter periods than a year or under suboptimal incubation conditions, the temperature and moisture modifiers need to be recalibrated for tropical conditions.

the mosquito Aedes aegypti vectors arboviruses such as urban yellow fever, dengue, chikungunya and Zika. Reducing the vector population is still the most effective method to decrease the spread of arboviruses. Fungi are the main natural disease agents of insects. The present study compared the virulence of conidia and blastospores of the entomopathogenic fungus Metarhizium anisopliae against A. aegypti pupae. The pupae were obtained by collecting eggs using "ovitraps" deployed in an urban environment. M. anisopliae conidia were produced using solid media and blastospores were produced by inoculating conidia in liquid culture. Blastospores were more virulent against pupae than conidia, and no pupae survived twenty-four hours after exposure to this type of propagule. Large quantities of mucilage were produced by the blastospores in the presence of the pupae. Exposure of pupae to conidial suspensions resulted in 57% survival at 24h and 23% at 48h. A proportion of the adults, which emerged from pupae exposed to conidia, succumbed to infection. This is the first study to demonstrates the infection of A. aegypti pupae by the entomopathogenic fungus M. anisopliae, showing that this stage of development can also be targeted by biological control agents.

Dengue is considered a serious public health problem in many tropical regions of the world including Brazil. At the moment, there is no viable alternative to reduce dengue infections other than controlling the insect vector, Aedes aegypti Linnaeus. In the continuing search for new sources of chemicals targeted at vector control, natural products are a promising alternative to synthetic pesticides. In our work, we investigated the toxicity of a bioactive compound extracted from the red alga Laurencia dendroidea J. Agardh. The initial results demonstrated that crude extracts, at a concentration of 5 ppm, caused pronounced mortality of second instar A. aegypti larvae. Two molecules, identified as (−)-elatol and (+)-obtusol were subsequently isolated from crude extract and further evaluated. Assays with (−)-elatol showed moderate larvicidal activity, whereas (+)-obtusol presented higher toxic activity than (−)-elatol, with a LC50 value of 3.5 ppm. Histological analysis of the larvae exposed to (+)-obtusol revealed damage to the intestinal epithelium. Moreover, (+)-obtusol-treated larvae incubated with 2 µM CM-H2DCFDA showed the presence of reactive oxygen species, leading us to suggest that epithelial damage might be related to redox imbalance. These results demonstrate the potential of (+)-obtusol as a larvicide for use against A. aegypti and the possible mode of action of this compound.