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Simple Summary: Previous studies have shown many populations of Culex quinquefasciatus are resistant to pyrethroids, the most common class of pesticide used by public health agencies. A genetic mutation has been identified that contributes to this insecticide resistance. Culex quinquefasciatus mosquitoes from numerous locations were tested to assess the correlation between standard resistance bioassays and this resistance mutation to determine if the mutation is a useful surrogate to assess insecticide resistance. The results from these Culex quinquefasciatus populations indicate that this kdr mutation is only a moderate strength correlate of phenotypic resistance and is thus unlikely to be a good surrogate for estimating insecticide resistance. Culex quinquefasciatus is an important target for vector control because of its ability to transmit pathogens that cause disease. Most populations are resistant to pyrethroids and often to organophosphates, the two most common classes of active ingredients used by public health agencies. A knockdown resistance (kdr) mutation, resulting in an amino acid change from a leucine to phenylalanine in the voltage gated sodium channel, is one mechanism contributing to the pyrethroid resistant phenotype. Enzymatic resistance has also been shown to play a very important role. Recent studies have shown strong resistance in populations even when kdr is relatively low, which indicates that factors other than kdr may be larger contributors to resistance. In this study, we examined, on a statewide scale (over 70 populations), the strength of the correlation between resistance in the CDC bottle bioassay and the kdr genotypes and allele frequencies. Spearman correlation analysis showed only moderate (−0.51) or weak (−0.29) correlation between the kdr genotype and permethrin or deltamethrin resistance, respectively. The frequency of the kdr allele was an even weaker correlate than genotype. These results indicate that assessing kdr in populations of Culex quinquefasciatus is not a good surrogate for phenotypic resistance testing. [ABSTRACT FROM AUTHOR]

Background: The development of insecticide resistance in disease-vectoring mosquito species can lead to vector control failure and disease resurgence. However, insecticide applications remain an essential public health intervention. In Florida, insecticide resistance in Aedes aegypti, an anthropophilic mosquito species capable of transmitting dengue, chikungunya, and Zika virus, is a major concern. Understanding the location, scale, and driving factors of insecticide resistance can enhance the ability of vector control organizations to target populations effectively. Methods: We used previously collected data on frequencies of mutations that confer resistance to commonly used pyrethroid insecticides in Ae. aegypti specimens from 62 sites distributed across 18 counties in Florida. To determine the scale of clustering for the most resistant variant, we used a Ripley's K function. We also used a spatial scanning statistic technique to identify locations of clusters where higher than expected frequencies of susceptible or resistant mosquitoes occurred. We then tested for associations between landscape, demographic, and insecticide-use factors using a beta regression modelling approach and evaluated the effect of spatial lag and spatial error terms on overall explanatory power of these models. Results: The scale at which maximum clustering of the most resistant variant occurs is approximately 20 kilometers. We identified statistically significant clusters of genotypes associated with resistance in several coastal cities, although some of these clusters were near significant clusters of susceptible mosquitoes, indicating selection pressures vary at the local scale. Vegetation density, distance from roads, and pyrethroid-use by vector control districts were consistently significant predictors of knockdown resistance genotype frequency in the top-performing beta regression models, although pyrethroid use surprisingly had a negatively associated with resistance. The incorporation of spatial lags resulted in improvements to the fit and explanatory power of the models, indicating an underlying diffusion process likely explains some of the spatial patterns observed. Conclusions: The genetic mutations that confer resistance to pyrethroids in Ae. aegypti mosquitoes in Florida exhibit spatial autocorrelation and patterns that can be partially explained by landscape and insecticide-use factors. Further work at local scales should be able to identify the mechanisms by which these variables influence selection for alleles associated with resistance. [ABSTRACT FROM AUTHOR]

Recent outbreaks of locally transmitted dengue and Zika viruses in Florida have placed more emphasis on integrated vector management plans for Aedes aegypti (L.) and Aedes albopictus Skuse. Adulticiding, primarily with pyrethroids, is often employed for the immediate control of potentially arbovirus-infected mosquitoes during outbreak situations. While pyrethroid resistance is common in Ae. aegypti worldwide and testing is recommended by CDC and WHO, resistance to this class of products has not been widely examined or quantified in Florida. To address this information gap, we performed the first study to quantify both pyrethroid resistance and genetic markers of pyrethroid resistance in Ae. aegypti and Ae. albopictus strains in Florida. Using direct topical application to measure intrinsic toxicity, we examined 21 Ae. aegypti strains from 9 counties and found permethrin resistance (resistance ratio (RR) = 6-61-fold) in all strains when compared to the susceptible ORL1952 control strain. Permethrin resistance in five strains of Ae. albopictus was very low (RR<1.6) even when collected from the same containers producing resistant Ae. aegypti. Characterization of two sodium channel kdr alleles associated with pyrethroid-resistance showed widespread distribution in 62 strains of Ae. aegypti. The 1534 phenylalanine to cysteine (F1534C) single nucleotide polymorphism SNP was fixed or nearly fixed in all strains regardless of RR. We observed much more variation in the 1016 valine to isoleucine (V1016I) allele and observed that an increasing frequency of the homozygous V1016I allele correlates strongly with increased RR (Pearson corr = 0.905). In agreement with previous studies, we observed a very low frequency of three kdr genotypes, IIFF, VIFF, and IIFC. In this study, we provide a statewide examination of pyrethroid resistance, and demonstrate that permethrin resistance and the genetic markers for resistance are widely present in FL Ae. aegypti. Resistance testing should be included in an effective management program. [ABSTRACT FROM AUTHOR]

Background: In this semi-field study, a new polymer-enhanced deltamethrin formulation, K-Othrine® PolyZone, was compared to a standard deltamethrin product for residual activity against a susceptible strain of laboratory-reared Anopheles gambiae using standard WHO cone bioassays. Methods: Residual insecticide efficacy was recorded after exposure to metal, cement and wood panels maintained in experimental huts in sub-tropical environmental conditions in north central Florida, USA, and panels stored in a climate controlled chamber located at the Centers for Disease Control and Prevention, Georgia, USA. Conclusions: K-Othrine® PolyZone demonstrated 100% control on metal and cement panels 1 year post application and > 80% control on wood panels up to 6 mo. The new formulation should be considered for use in indoor residual spray programmes requiring long-term control of malaria vectors. [ABSTRACT FROM AUTHOR]

Sporadic outbreaks of human cases of West Nile virus (WNV), primarily vectored by Culex quinquefasciatus Say in suburban and urban areas, have been reported since introduction of the virus into Florida in 2001. Miami-Dade County, Florida is part of one of the largest metropolitan areas in the United States, supports Cx. quinquefasciatus year-round, and recently experienced over 60 human cases of WNV during one outbreak. To facilitate more effective integrated vector management and public health protection, we used the Centers for Disease Control and Prevention (CDC) bottle bioassay method to evaluate the susceptibility of adult Cx. quinquefasciatus collected from 29 locations throughout Miami-Dade County to pyrethroid and organophosphate adulticide active ingredients (AIs) used by Miami-Dade County Mosquito Control. We also determined the frequency of the 1014 knockdown resistance (kdr) mutation for Cx. quinquefasciatus from a subset of 17 locations. We detected resistance to two pyrethroid AIs in all tested locations (permethrin: 27 locations, deltamethrin: 28 locations). The 1014F allele was widely distributed throughout all 17 locations sampled; however, 29.4% of these locations lacked 1014F homozygotes even though phenotypic pyrethroid resistance was present. Organophosphate resistance was more variable; 20.7% of the locations tested were susceptible to malathion, and 33.3% of the populations were susceptible to naled. We subsequently conducted a field trial of ReMoa Tri, a recently approved multiple AI adulticide formulation labelled for resistant mosquitoes, against a mixed location field population of Miami-Dade Cx. quinquefasciatus. Average 24-hr mortality was 65.1 ± 7.2% and 48-hr mortality increased to 85.3 ± 9.1%, indicating good control of these resistant Cx. quinquefasciatus. This current study shows that insecticide resistance is common in local Cx. quinquefasciatus but effective options are available to maintain control during active disease transmission in Miami-Dade County. [ABSTRACT FROM AUTHOR]

Mosquitoes have a wide range of digestive enzymes that enable them to utilize requisite blood and sugar meals for survival and reproduction. Sugar meals, typically derived from plant sources, are critical to maintain energy in both male and female mosquitoes, whereas blood meals are taken only by females to complete oogenesis. Enzymes involved in sugar digestion have been the subject of study for decades but have been limited to a relatively narrow range of mosquito species. The southern house mosquito, Culex quinquefasciatus, is of public health importance and seldom considered in these types of studies outside of topics related to Bacillus sphaericus, a biocontrol agent that requires interaction with a specific gut-associated α-glucosidase. Here we sought to describe the nature of α-glucosidases and unexplored β-glucosidases that may aid Cx. quinquefasciatus larvae in acquiring nutrients from cellulosic sources in their aquatic habitats. Consistent with our hypothesis, we found both α- and β-glucosidase activity in larvae. Interestingly, β-glucosidase activity all but disappeared at the pupal stage and remained low in adults, while α-glucosidase activity remained in the pupal stage and then exceeded larval activity by approximately 1.5-fold. The expression patterns of the putative α- and β-glucosidase genes chosen did not consistently align with observed enzyme activities. When the α-glucosidase inhibitor acarbose was administered to adults, mortality was seen especially in males but also in females after two days of exposure and key energetic storage molecules, glycogen and lipids, were significantly lower than controls. In contrast, administering the β-glucosidase inhibitor conduritol β-epoxide to larvae did not produce mortality even at the highest soluble concentration. Here we provide insights into the importance of α- and β-glucosidases on the survival of Cx. quinquefasciatus in their three mobile life stages. [ABSTRACT FROM AUTHOR]

A growing number of investigations of essential oils, extracts or phytochemicals have been conducted to discover effective pesticidal agents against Aedes aegypti L., a major vector of yellow fever, Zika fever and dengue. The goal of this study was to enhance the knowledge about the activity of Erica manipuliflora and to discover effective pesticidal agents against Ae. aegypti L during both the flowering and fruiting periods. Eight extracts of E. manipuliflora in flowering and fruiting periods were investigated for their insecticidal activities against larval and adult Ae. aegypti. Five extracts exhibited adult mortality of greater than 90% at 5 μg/mosquito. The major volatile compounds of petroleum ether and acetone extracts of the aerial parts in both periods were found cyclopentanol, methylcyclohexane, 2-methyl-2-pentanol and 4-hydroxy-4-methyl-1,2-pentanone respectively, by GC-MS. 5- O -caffeoylquinic acid, myricetin rhamnoside, and quercetin rhamnoside were determined as major compounds of active extracts by LC-MS. This initial study of the insecticidal activity of E. manipuliflora shows it has compounds potentially useful for protection against vector-borne diseases. [Display omitted] • This is the first study on insecticidal activity of Erica manipuliflora extracts. • The activity results were evaluated statistically. • The contents of active extracts were analyzed by different chromatography methods. • It has a good potential for protection against vector-borne diseases. [ABSTRACT FROM AUTHOR]

The major family of neuropeptides (NPs) derived from the pk (pyrokinin)/ pban (pheromone biosynthesis activating neuropeptide) gene are defined by a common FXPRL-NH 2 or similar sequence at the C-termini. This family of peptides has been found in all insect groups investigated to date and is implicated in regulating various physiological functions, including pheromone biosynthesis and diapause, but other functions are still largely unknown in specific life stages. Here we identify two isoforms of pk / pban cDNA encoding the PBAN domain from the sand fly Phlebotomus papatasi . The two pk / pban isoforms have the same sequence except for a 63 nucleotide difference between the long and short forms, and contain no alternative mRNA splicing site. Two NP homologues, DASGDNGSDSQRTRPPFAPRLamide and SLPFSPRLamide are expected, however, sequence corresponding to the diapause hormone was not found in the P. papatasi pk / pban gene. The PBAN-like amino acid sequence homologue SNKYMTPRL is conserved in the gene, but there is no cleavage site for processing a functional peptide. Characterizing the expression of the isoforms in developmental stages and adults indicates that the short form is differentially transcribed depending on the life stage. The P. papatasi pk / pban gene is the only known pk / pban gene with two transcriptional isoforms and from examination of endoproteolytic cleavage sites is expected to produce fewer peptides than most of the pk / pban genes elucidated to date; only Drosophila melanogaster is simpler with a single NP detected by mass spectroscopy. A phylogenetic analysis showed P. papatasi pk / pban grouped more closely with other nematoceran flies rather than higher flies. [ABSTRACT FROM AUTHOR]

Culex pipiens complex is an important vector of epizootic and zoonotic pathogens, including West Nile virus. Chicago, Illinois and its suburbs have suffered high incidence of human West Nile virus infections in the past. This makes abatement programs in and around the Chicago area an essential service. The control of Cx. pipiens is often complicated by rapidly evolving resistance to pyrethroids, which are the most widely used chemical class in US mosquito abatement programs. The present study assessed Sumithrin® resistance in Cx. pipiens collected from five locations around Cook County, Illinois, neighboring the city limits of Chicago. According to CDC guidelines, samples from all five locations demonstrated some resistance to Sumithrin®. When assessed with Anvil®, a formulated product made of Sumithrin® synergized with piperonyl butoxide, susceptibility was rescued in mosquitoes from three out of the five locations, suggesting involvement of mixed-function oxidases and/or carboxylesterases in Sumithrin® resistance at these locations. Not all locations had susceptibility rescued by Anvil®, but these locations had relatively low knockdown resistance allele frequencies, suggesting that mechanisms other than knockdown resistance may be involved. Enzyme activities did not reveal any marked trends that could be related back to mortality in the bottle bioassays, which highlights the need for multiple types of assays to infer enzymatic involvement in resistance. Future directions in pyrethroid resistance management in Chicago area Cx. pipiens are discussed. [ABSTRACT FROM AUTHOR]

• Insecticidal activity of 47 macroalgae mostly from Mediterranean Sea was examined. • Cholinesterase and tyrosinase inhibitory effect of the alga extracts was investigated. • D. dichotoma var. intricata was the only one exhibiting BChE inhibition over 50%. • BChE inhibition by four diterpenes in DDI was investigated in silico. • The macroalgae and their metabolites might be new sources of novel insecticides. Ethanolic extracts prepared from forty-seven macroalgae species collected from the Mediterranean Sea, Aegean Sea, and Sea of Marmara were tested at 200 μg/mL against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase (TYR), crucial for insect vivacity. Only one alga extract, e.g. Dictyota dichotoma var. intricata (C.Agardh) Greville 1830, had a notable inhibition of BChE (72.0 ± 0.07%), whereas the rest of the macroalgae possessed no or low inhibition of AChE (2.20 ± 0.39% - 14.20 ± 2.16%), BChE (0.62 ± 0.07% - 41.20 ± 3.07%), and TYR (< 10%). Several known diterpenes (isoamijiol, 14-deoxyamijiol, amijidictyol, dictyodial, and 4α-acetoxydictyodial) found earlier in this species were proceeded to molecular docking experiments to figure out its interactions with BChE. Eleven of the 47 macroalgal extracts were selected based on sufficient availability for testing in larvicidal and adulticidal activity assays against female Aedes aegypti "Orlando1952" the mosquito vector of yellow fever and dengue. The selected extracts displayed a high adulticidal activity ranging between 80–100% activity at 5 μg/mosquito. In contrast, only the ethanol extract of Ceramium siliquosum var. elegans showed larvicidal mortality of 93.3 ± 11.5% at 0.5 μg/mL. Our findings show that the macroalgae and their metabolites could be sources of novel insecticidal agents. [ABSTRACT FROM AUTHOR]

• The volatile compounds of its roots were analyzed by GC and GC-MS analysis. • The insecticidal activity of V.dioscoridis extracts was reported for the first time. • The cytotoxicity (HepG2 and HUVEC cells) of its extracts reported for the first time. • The activity results were evaluated by one-way ANOVA and Tukey's post hoc test. • The selective toxicity against cancer cells were evaluated with Selectivity index. Zika, dengue, and Yellow fever are transmitted primarily by Aedes aegypti mosquitoes. Investigations to find more natural but highly effective pesticidal agents against Ae. aegypti have increased in the last few years. Plant-based compounds have been the source of several current pesticides and have provided models for creation of synthetic derivatives like the pyrethroids. Plants continue to be a rich source for potential new active ingredients. The aim of this study is to increase knowledge on V. dioscoridis and to present the results of the cytotoxic and insecticidal activities of various extracts of its roots and volatile compound compositions of roots. The hexane and chloroform extracts, HM1 and CM1 respectively, exhibited cytotoxicity with IC 50 values of 128.4 and 86.93 µg/mL against the HepG2 cell line. Because CM1 had higher cytotoxicity across the studied concentration range, it was also tested against the HUVEC cell line. Extracts HM1 and CM1 were also tested for their insecticidal activities. CM1 was found to be the more active (mortality (%) = 96.7% ± 5.8 at 5 μg/mosquito), consistent with its cytotoxic activity. Since these active extracts were nonpolar, the chemical composition of the volatile compounds of the roots were analyzed by GC-MS analysis. About 87.7% of the essential oil were characterized. The major compound was ß-eudesmol (9.2%), following by camphene (8.9%), bornyl acetate (8.2%) and maaliol (7.6%). This is the first study on the HepG2 and HUVEC cytotoxicity and insecticidal activity of V. dioscoridis. Further studies on the potential of this plant to define the spectrum of activity and examine potential therapeutic agents is warranted. [ABSTRACT FROM AUTHOR]

Rickettsiae and bartonellae are Gram-negative bacteria that can cause zoonotic and human diseases and are vectored by hematophagous arthropods. In the Americas, rickettsioses and bartonelloses have reemerged as significant public health threats. Bartonella species have been identified as causing zoonotic infections responsible for a variety of clinical syndromes in humans and animals. The aim of this study was to investigate the distribution, prevalence, and molecular heterogeneity of Rickettsia spp. and Bartonella spp. among ectoparasites collected from domestic animals in 14 farming communities in the Andes Mountains of Cuzco, Peru. A total of 222 domestic animals representing 8 different species (sheep, donkeys, goats, cattle, pigs, llamas, guinea pigs, and horses) were sampled. Nine species of ectoparasites (n = 1,697) collected from 122 animals were identified resulting in 1,657 chewing lice, 39 ticks, and 1 flea. DNA was individually extracted from a random sample of 600 (35.4%) considering variability of ectoparasite species, hosts, and sample location elevation. All 600 samples were negative for rickettsial DNA by a genus-specific molecular assay. A subset of 173 (28.8%) samples were selected based on variability of arthropods species, host, and location for Bartonella testing. Ninety-one (52.6%) of these samples including Melophagus ovinus (90/110) and Bovicola bovis (1/7) were positive for Bartonella by a genus-specific molecular assay. Five Bartonella genes of seven DNA samples from M. ovinus were analyzed by the multilocus sequence typing for characterization. We identified five identical Bartonella melophagi specimens and two specimens with Bartonella species related to B. melophagi from the seven M. ovinus. The Bartonella agents detected were widely distributed and frequent in multiple studied locations. [ABSTRACT FROM AUTHOR]

In this study, Magnolia citrata Noot and Chalermglin (Magnoliaceae) essential oil (MCEO) was evaluated for insecticidal activity against the yellow fever mosquito Aedes aegypti and attractant activity for the Mediterranean fruit fly Ceratitis capitata. The leaves of Magnolia citrata (Giổi chanh) were collected from northwestern Vietnam, and the water-distilled MCEO was analyzed by gas-chromatography and mass spectrometry (GC-MS). The major constituents of MCEO were identified as linalool 19%, geranial 16%, citronellal 14%, neral 14%, and sabinene 12%. MCEO showed 100% mortality at 1 μg/μL against 1st instar larvae of Ae. aegypti (Orlando strain, ORL), and the oil exhibited 54% (ORL) and 68% (Puerto Rico strain) mortality at 5 μg/mosquito against Ae. aegypti adult females. Initial screens showed that MCEO had weak insecticidal activity compared to the positive control permethrin. In bioassays with sterile male C. capitata, MCEO exhibited moderately strong attraction, comparable to that observed with a positive control, Tetradenia riparia essential oil (TREO). Herein, the insecticidal and attractant activities of MCEO are reported for the first time. [ABSTRACT FROM AUTHOR]

The adulticidal, larvicidal, and repellent activity of 18 trifluoromethylphenyl amides (TFMPAs) was determined against Aedes aegypti mosquitoes. The compounds studied are the third generation designed from active structures of the previous two generations. N -(3,5-Bis(trifluoromethyl)phenyl)-2-chloroacetamide (8f) and N -(3,5-bis(trifluoromethyl)phenyl)-2,2,3,3,3-pentafluoropropanamide (8h) were most active against 1st stage Ae. aegypti larvae with LC 50 values of 125 and 2.53 μM; for comparative purposes, the published LC 50 for fipronil is 0.014 μM. Compound 8h was the most toxic against adult female Ae. aegypti with an LD 50 = 2.12 nmol/mg, followed by 8f, and N -(3,5-bis(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (8g) with LD 50 values of 4.27 and 4.73 nmol/mg, respectively, although these compounds were significantly less toxic than fipronil against adult female Ae. aegypti. Compounds N -(2-(trifluoromethyl)phenyl)butyramide (9c), N -(2-(trifluoromethyl)phenyl)pentanamide (9d) and N -(2-(trifluoromethyl)phenyl)hex-5-enamide (9e) were the best repellents for female Ae. aegypti, with minimum effective dosages (MEDs) of 0.026, 0.052, and 0.091 μmol/cm2 , respectively, compared to DEET at 0.052 μmol/cm2. Out of 52 TFMPAs (total number of compounds from three generations of this research) compound 9c was the most active repellent along with two synthesized in our previous studies, 2-chloro- N -(3-(trifluoromethyl)phenyl)acetamide (6a) and 2,2,2-trifluoro- N -(2-(trifluoromethyl)phenyl)acetamide (4c). Unlabelled Image • Compounds were tested against Ae. aegypti mosquitoes for toxicity and repellency. • Some compounds had adulticidal/larvicidal activity, but they were not as active as fipronil. • Three compounds showed repellency equal to or better than the DEET standard. [ABSTRACT FROM AUTHOR]

Valeriana is a common plant species used for various healing purposes in folk medicine since antiquity. This study investigates the phytochemical profile, antioxidant, cytotoxic, and insecticidal activity of Valeriana alliariifolia Adams, a species that has traditionally been used in Turkey. For the analyses we prepared four root extracts of V. alliariifolia Adams using hexane (HM1), chloroform (CM1), ethanol (EM1), and water (WM1) for maceration. Additionally, two extracts were also prepared from its roots by maceration separately with ethanol (EM2) and water (WM2). One sample was prepared as a water infusion (WI), according to the procedure used in Turkish traditional medicine. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical cation scavenging activity tests showed that ethanol extracts had the strongest antioxidant activity: EM1 (IC50 – DPPH: 17.694 µg/mL; ABTS: 23.8 µg/mL) and EM2 (IC50 – DPPH: 20 µg/mL; ABTS: 21.5 µg/mL). The hexane extract, HM1, was the most cytotoxic (IC50<10 µg/mL against HepG2 and HUVEC) and EM2 strongly cytotoxic (IC50<10 µg/mL against HepG2 and IC50: 11.96 µg/mL against HUVEC). The extracts with demonstrated cytotoxic activities were further examined to check their insecticidal activity against adult female mosquito Aedes aegypti and first instar Ae. aegypti larvae. HM1 was the most effective (90±10 %), which was consistent with its cytotoxic activity. Because of the high antioxidant, cytotoxic, and insecticidal activities, we ran phytochemical analyses of the HM1, EM1, and EM2 extracts with GC-MS (for HM1) and LC-MS/MS (for EM1 and EM2). We also analysed the composition of the essential oil obtained from V. alliariifolia roots by micro-distillation in order to compare its content with HM1, which contains volatile compounds. Phytochemical analyses revealed that the major compound in HM1 was isovaleric acid (16 %) and in the essential oil 1,8-cineole (2.9 %). EM1 and EM2 contained 5-O-caffeoylquinic acid (chlorogenic acid), verbascoside (acteoside), and 3,5-dicaffeoylquinic acid as major components. In the light of our findings and available literature, we can conclude that V. alliariifolia has a good bioactive potential that could be used for different purposes, including the development of new agents for the treatment of various diseases. The difference in the content between the essential oil and HM1 was remarkable. It suggests that the variability observed in the activity of the samples was a result of composition and that, therefore, the aim of treatment should dictate which type of preparation is to be selected. An added value of our study is that it determined verbascoside and methylquercetin rutinoside for the first time in the Valeriana extracts. [ABSTRACT FROM AUTHOR]

Abstract This project focused on the design, synthesis, and testing of trifluoromethylphenyl amides (TFMPAs) as potential mosquitocides and repellents. Fourteen compounds were evaluated for toxicity against larvae and adults of Aedes aegypti. Several compounds were toxic against Aedes aegypti (larval, adult and feeding bioassays) and Drosophila melanogaster (glass-surface contact assay), but were much less toxic than fipronil, with toxicity ratios ranging from 100-fold in the larval assay to 100,000-fold for topical application to adult insects. In repellency bioassays to determine minimum effective dosage (MED), compound N -(2,6-dichloro-4-(trifluoromethyl)phenyl)-2,2,3,3,3-pentafluoropropanamide (7b) repelled Ae. aegypti females at lower concentration, 0.017 (±0.006) μmol/cm2, than N , N -diethyl-meta-toluamide (DEET) 0.026 (±0.005) μmol/cm2. 2-Chloro- N -(3-(trifluoromethyl)phenyl)acetamide (6a) performed better than DEET against two species of mosquitoes: it repelled Ae. aegypti females at 0.013 (±0.006) μmol/cm2 and Anopheles gambiae females (in a warm body repellent assay), at a standard exposure of 2 nmol/cm2. These studies revealed novel active structures that could further lead to compounds with better repellent activity. Graphical abstract Unlabelled Image Highlights • A total of 14 trifluoromethylphenyl amides (4 novel) were synthesized. • Compounds were evaluated for insecticidal and repellent activity. • Insecticidal activity was low, but one compound was a better repellent than DEET • Compounds from this study may be used to design novel repellents. [ABSTRACT FROM AUTHOR]

Abstract: We describe a unique microsporidian species that infects the green stink bug, Chinavia hilaris; the brown marmorated stink bug, Halyomorpha halys; the brown stink bug, Euschistus servus; and the dusky stink bug, Euschistus tristigmus. All life stages are unikaryotic, but analysis of the consensus small subunit region of the ribosomal gene places this microsporidium in the genus Nosema, which historically has been characterized by diplokaryotic life stages. It is also characterized by having the reversed arrangement of the ribosomal gene (LSU –ITS‐ SSU) found in species within the “true Nosema” clade. This microsporidium is apparently Holarctic in distribution. It is present in H. halys both where it is native in Asia and where it is invasive in North America, as well as in samples of North American native C. hilaris collected prior to the introduction of H. halys from Asia. Prevalence in H. halys from mid‐Atlantic, North America in 2015–2016 ranged from 0.0% to 28.3%, while prevalence in C. hilaris collected in Illinois in 1970–1972 ranged from 14.3% to 58.8%. Oral infectivity and pathogenicity were confirmed in H. halys and C. hilaris. Morphological, ultrastructural, and ecological features of the microsporidium, together with a molecular phylogeny, establish a new species named Nosema maddoxi sp. nov. [ABSTRACT FROM AUTHOR]

House flies (Musca domestica) are worldwide agricultural pests with estimated control costs at $375 million annually in the U.S. Non-target effects and widespread resistance challenge the efficacy of traditional chemical control. Double stranded RNA (dsRNA) has been suggested as a biopesticide for M. domestica but a phenotypic response due to the induction of the RNAi pathway has not been demonstrated in adults. In this study female house flies were injected with dsRNA targeting actin-5C or ribosomal protein (RP) transcripts RPL26 and RPS6. Ovaries showed highly reduced provisioning and clutch reductions of 94–99% in RP dsRNA treated flies but not in actin-5C or GFP treated flies. Gene expression levels were significantly and specifically reduced in dsRNA injected groups but remained unchanged in the control dsGFP treated group. Furthermore, injections with an Aedes aegypti conspecific dsRNA designed against RPS6 did not impact fecundity, demonstrating species specificity of the RNAi response. Analysis of M. domestica tissues following RPS6 dsRNA injection showed significant reduction of transcript levels in the head, thorax, and abdomen but increased expression in ovarian tissues. This study demonstrates that exogenous dsRNA is specifically effective and has potential efficacy as a highly specific biocontrol intervention in adult house flies. Further work is required to develop effective methods for delivery of dsRNA to adult flies. [ABSTRACT FROM AUTHOR]

Aedes albopictus Skuse (Diptera: Culicidae) was tested for resistance to permethrin, bifenthrin, and malathion using Centers for Disease Control and Prevention (CDC) bottle bioassays and topical toxicology assays on adults and larval bioassays. Eggs were collected from 3 locations across St. Johns County, Florida, raised to the F3 generation and compared with an insecticide susceptible laboratory strain. Results from CDC bottle bioassays with permethrin indicate no significant differences between the 3 wild-type strains and the laboratory strain but suggest the possibility of resistance in 1 strain. Bottle bioassay results for malathion were inconclusive. Topical toxicological results for adults and bioassays for larvae showed a significant difference in permethrin resistance between the control strain and 1 of the wild-type strains. Results from this project indicate that insecticide susceptibility testing should be a regular part of mosquito surveillance programs. Upon detection of resistance, detailed dose response bioassays should be performed to quantify the resistance and mechanisms in local vector populations. [ABSTRACT FROM AUTHOR]

Metaseiulus occidentalis is an eyeless phytoseiid predatory mite employed for the biological control of agricultural pests including spider mites. Despite appearances, these predator and prey mites are separated by some 400 Myr of evolution and radically different lifestyles. We present a 152-Mb draft assembly of the M. occidentalis genome: Larger than that of its favored prey, Tetranychus urticae, but considerably smaller than those ofmany other chelicerates, enabling an extremely contiguous and complete assembly to be built-the best arachnid to date. Aided by transcriptome data, genome annotation cataloged 18,338 protein-coding genes and identified large numbers of Helitron transposable elements. Comparisonswith other arthropods revealed a particularly dynamic and turbulent genomic evolutionary history. Its genes exhibit elevated molecular evolution, with strikingly high numbers of intron gains and losses, in stark contrast to the deer tick Ixodes scapularis.Uniquely among examined arthropods, this predatorymite's Hox genes are completely atomized, dispersed across the genome, and it encodes five copies of the normally single-copy RNAprocessingDicer-2 gene. Examining gene families linked to characteristic biological traits of this tiny predator provides initial insights into processes of sex determination,development,immunedefense,andhowitdetects,disables, anddigests its prey.As thefirst referencegenome for the Phytoseiidae, and for any species with the rare sex determination system of parahaploidy, the genome of the western orchard predatorymite improves genomic sampling of chelicerates and provides invaluable newresources for functional genomic analyses of this family of agriculturally important mites. [ABSTRACT FROM AUTHOR]

An undescribed microsporidium was detected and isolated from the South American bumble bee Bombus atratus collected in the Pampas region of Argentina. Infection intensity in workers averaged 8.2 × 10 7 spores/bee. The main site of infection was adipose tissue where hypertrophy of adipocytes resulted in cyst-like body formation. Mature spores were ovoid and monomorphic. They measured 4.00 μm × 2.37 μm (fresh) or 3.98 μm × 1.88 μm (fixed). All stages were diplokariotic and developed in direct contact with host cytoplasm. Isofilar polar filament was arranged in 16 coils in one or, posteriorly, two layers. Coiling angle was variable, between perpendicular and almost parallel to major spore axis. Late meronts and sporogonial stages were surrounded by vesicles of approximately 60 nm in diameter. Based on both new and already designed primers, a 1827 bp (SSUrRNA, ITS, LSUrRNA) sequence was obtained. Data analyses suggest that this microsporidium is a new species of the genus Tubulinosema. The name Tubulinosema pampeana sp. n. is proposed. [ABSTRACT FROM AUTHOR]

The Testing and Evaluation Department of the US Navy Entomology Center of Excellence (NECE), Naval Air Station, Jacksonville, Florida, is dedicated to the evaluation of novel equipment and vector control techniques to provide guidance on effective protection measures against human pathogens transmitted by blood-feeding arthropods. Personal protective measures (PPM), to include repellents, are part of a series of techniques that contribute toward reducing human-vector contact for globally and domestically deployed military forces. However, improper PPM use and limited availability has created vulnerabilities, causing troops to purchase spatial repellent products that are not approved by the Department of Defense. In order to ensure the most effective products are available, NECE has evaluated the spatial repellency response of Aedes albopictus (Skuse) to 4 commercial off-the-shelf (COTS) spatial repellents to provide technical guidance on proper use and effectiveness. The COTS products evaluated ThermaCELL, OFF! Clip On, Lentek Bite Shield, and Bug Button Mosquito Eliminator. A Biogents Sentinel (BGS) trap was placed in 5 locations with a spatial repellent device suspended at the level of the BGS trap opening over 4 of them (the fifth was control). Each trap catch was collected every 12 hours, at which time the spatial repellent device was rotated to the next position. Using this method, each spatial repellent device and control was rotated across each of the 5 locations a total of 6 times. Spatial repellent efficiency was evaluated by comparing the total number of mosquitoes collected in the BGS traps during a 12-hour period. The number of adult mosquitoes repelled by the ThermaCell spatial repellent was significantly more than other spatial repellents with the exception of OFF!. These data indicate that COTS products using repellent insecticide rather than botanicals are more effective at deterring Ae. albopictus from biting a host. [ABSTRACT FROM AUTHOR]

Chromatographic purification of the alcoholic extract from the aerial parts of the Saudi plant Nuxia oppositifolia (Hochst.), Benth., resulted in five isolated phenolic compounds. Two flavones, hispidulin (1) and jaceosidin (2), and the phenylethanoid glycosides, verbascoside (3), isoverbascoside (4), and conandroside (5), were identified and their chemical structures were determined by spectroscopic analyses. The insecticidal activity of compounds 1 and 2, in addition to 11 compounds isolated in a previous research (6–16), was evaluated against the Yellow Fever mosquito, Aedes aegypti. Four compounds displayed adulticidal activity with LD50 values of 2–2.3 μg/mosquito. Free radical scavenging properties of the plant extracts and compounds (1–5) were evaluated by measuring the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonate radical cation (ABTS•+) scavenging activity. All compounds exhibited notable activity, compared with the positive control, l-Ascorbic acid. This study suggests that N. oppositifolia could be a promising source of secondary metabolites, some with lethal adulticidal effect against Ae. aegypti. [ABSTRACT FROM AUTHOR]