scholarly journals Oxidase, Esterase, and KDR-Associated Pyrethroid Resistance in Culex quinquefasciatus Field Collections of Collier County, Florida

2020 ◽  
Vol 36 (1) ◽  
pp. 22-32
Author(s):  
Keira J. Lucas ◽  
Rachel B. Bales ◽  
Kaci McCoy ◽  
Caroline Weldon

ABSTRACT In several insect species, resistance to pyrethroids and DDT (dichlorodiphenyltrichloroethane) is linked to point mutations in the voltage-gated sodium channel (VGSC) gene. Pyrethroid-based insecticides prolong the opening of sodium channels, causing paralysis known as a “knockdown” effect before mortality occurs. Point mutations in the VGSC gene result in decreased pyrethroid binding and reduced sensitivity to the insecticide—this resistance mechanism is known as knockdown resistance (kdr) as insects do not die but recover from paralysis with time. In Culex mosquito species loss of target site sensitivity to pyrethroids is linked to a number of substitutions, one of which is leucine (L) to phenylalanine (F) at residue 1014 (L1014F) in the VGSC gene. Here we report the identification of kdr-associated pyrethroid resistance and developing resistance in Cx. quinquefasciatus field collections from Collier County, FL. Evaluation of position 1014 of the VGSC in Cx. quinquefasciatus collections from 7 locations in Collier County, FL, revealed a wide range of genotypes from one part of the district to the other. Centers for Disease Control and Prevention bottle bioassay, linear regression analysis, and cage trial evaluations suggest that the L1014F mutation plays a role, at least in part, to the pyrethroid resistance status of Cx. quinquefasciatus collected in Collier County, FL. Furthermore, we identified resistance attributed to both oxidase and esterase activity, indicating that multiple mechanisms are responsible for pyrethroid resistance in Collier County Cx. quinquefasciatus.

2012 ◽  
Vol 109 (38) ◽  
pp. 15206-15211 ◽  
Author(s):  
Nicole Joußen ◽  
Sara Agnolet ◽  
Sybille Lorenz ◽  
Sebastian E. Schöne ◽  
Renate Ellinger ◽  
...  

Worldwide, increasing numbers of insects have evolved resistance to a wide range of pesticides, which hampers their control in the field and, therefore, threatens agriculture. Members of the carboxylesterase and cytochrome P450 monooxygenase superfamilies are prominent candidates to confer metabolic resistance to pyrethroid insecticides. Both carboxylesterases and P450 enzymes have been shown to be involved in pyrethroid resistance in Australian Helicoverpa armigera, the noctuid species possessing by far the most reported resistance cases worldwide. However, specific enzymes responsible for pyrethroid resistance in field populations of this species have not yet been identified. Here, we show that the resistance toward fenvalerate in an Australian strain of H. armigera is due to a unique P450 enzyme, CYP337B3, which arose from unequal crossing-over between two parental P450 genes, resulting in a chimeric enzyme. CYP337B3 is capable of metabolizing fenvalerate into 4′-hydroxyfenvalerate, which exhibits no toxic effect on susceptible larvae; enzymes from the parental P450 genes showed no detectable fenvalerate metabolism. Furthermore, a polymorphic H. armigera strain could be bred into a susceptible line possessing the parental genes CYP337B1 and CYP337B2 and a resistant line possessing only CYP337B3. The exclusive presence of CYP337B3 in resistant insects of this strain confers a 42-fold resistance to fenvalerate. Thus, in addition to previously documented genetic mechanisms of resistance, recombination can also generate selectively advantageous variants, such as this chimeric P450 enzyme with an altered substrate specificity leading to a potent resistance mechanism.


2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Lorena I. Orjuela ◽  
Diego A. Álvarez-Diaz ◽  
Juliana A. Morales ◽  
Nelson Grisales ◽  
Martha L. Ahumada ◽  
...  

Abstract Background Knockdown resistance (kdr) is a well-characterized target-site insecticide resistance mechanism that is associated with DDT and pyrethroid resistance. Even though insecticide resistance to pyrethroids and DDT have been reported in Anopheles albimanus, Anopheles benarrochi sensu lato (s.l.), Anopheles darlingi, Anopheles nuneztovari s.l., and Anopheles pseudopunctipennis s.l. malaria vectors in Latin America, there is a knowledge gap on the role that kdr resistance mechanisms play in this resistance. The aim of this study was to establish the role that kdr mechanisms play in pyrethroid and DDT resistance in the main malaria vectors in Colombia, in addition to previously reported metabolic resistance mechanisms, such as mixed function oxidases (MFO) and nonspecific esterases (NSE) enzyme families. Methods Surviving (n = 62) and dead (n = 67) An. nuneztovari s.l., An. darlingi and An. albimanus mosquitoes exposed to diagnostic concentrations of DDT and pyrethroid insecticides were used to amplify and sequence a ~ 225 bp fragment of the voltage-gated sodium channels (VGSC) gene. This fragment spanning codons 1010, 1013 and 1014 at the S6 segment of domain II to identify point mutations, which have been associated with insecticide resistance in different species of Anopheles malaria vectors. Results No kdr mutations were detected in the coding sequence of this fragment in 129 samples, 62 surviving mosquitoes and 67 dead mosquitoes, of An. darlingi, An. nuneztovari s.l. and An. albimanus. Conclusion Mutations in the VGSC gene, most frequently reported in other species of the genus Anopheles resistant to pyrethroid and DDT, are not associated with the low-intensity resistance detected to these insecticides in some populations of the main malaria vectors in Colombia. These results suggest that metabolic resistance mechanisms previously reported in these populations might be responsible for the resistance observed.


2021 ◽  
Vol 68 (1) ◽  
pp. 14-23
Author(s):  
Alexandria S. Watkins

The Naples Botanical Garden, located in Collier County, Florida, attracts over 220,000 visitors each year. The gardens house a collection of plants from around the world, including a featured area for over 100 species of exotic and native bromeliads. Ornamental bromeliads have previously been investigated to define their “tank” structure as a haven for mosquito eggs and larvae. The Naples Botanical Gardens was investigated for the presence of juvenile mosquitoes inhabiting large-tanked bromeliads. A survey of mosquito species inhabiting bromeliads in the gardens indicated that the most abundant species was Culex quinquefasciatus. With the ongoing threat of vector borne diseases such as West Nile virus, the abundance of vector mosquitoes and heavy tourist traffic in the gardens, insecticide resistance testing was performed on Cx. quinquefasciatus originating in the gardens in order to assess the ability of pyrethroid-based insecticides used by the local vector control agency to successfully target this species in the event of a disease outbreak. We identified pyrethroid resistance in Cx. quinquefasciatus collected from Naples Botanical Gardens, and that oxidase activity was the primary mechanism responsible for its pyrethroid resistance status.


Insects ◽  
2022 ◽  
Vol 13 (1) ◽  
pp. 92
Author(s):  
Joanna M. Reinhold ◽  
Karthikeyan Chandrasegaran ◽  
Helen Oker ◽  
José E. Crespo ◽  
Clément Vinauger ◽  
...  

Combining thermopreference (Tp) and CO2-gated heat-seeking assays, we studied the thermal preferendum and response to thermal cues in three Culex mosquito species exhibiting differences in native habitat and host preference (e.g., biting cold and/or warm-blooded animals). Results show that these species differ in both Tp and heat-seeking behavior. In particular, we found that Culex territans, which feed primarily on cold-blood hosts, did not respond to heat during heat-seeking assays, regardless of the CO2 concentration, but exhibited an intermediate Tp during resting. In contrast, Cx. quinquefasciatus, which feeds on warm blooded hosts, sought the coolest locations on a thermal gradient and responded only moderately to thermal stimuli when paired with CO2 at higher concentrations. The third species, Cx. tarsalis, which has been shown to feed on a wide range of hosts, responded to heat when paired with high CO2 levels and exhibited a high Tp. This study provides the first insights into the role of heat and CO2 in the host seeking behavior of three disease vectors in the Culex genus and highlights differences in preferred resting temperatures.


2021 ◽  
pp. 096100062110165
Author(s):  
Mohammadhiwa Abdekhoda ◽  
Fatemeh Ranjbaran ◽  
Asghar Sattari

This study was conducted with the aim of evaluating the role of information and information resources in the awareness, control, and prevention of COVID-19. This study was a descriptive-analytical survey in which 450 participants were selected for the study. The data collection instrument was a researcher-made questionnaire. Descriptive and inferential statistics were used to analyze the data through SPSS. The findings show that a wide range of mass media has become well known as information resources for COVID-19. Other findings indicate a significant statistical difference in the rate of using information resources during COVID-19 based on age and gender; however, this difference is not significant regarding the reliability of information resources with regard to age and gender. Health information has an undisputable role in the prevention and control of pandemic diseases such as COVID-19. Providing accurate, reliable, and evidence-based information in a timely manner for the use of resources and information channels related to COVID-19 can be a fast and low-cost strategic approach in confronting this disease.


Genetics ◽  
2001 ◽  
Vol 159 (3) ◽  
pp. 1103-1115 ◽  
Author(s):  
Hongguang Shao ◽  
Zhijian Tu

Abstract A novel transposon named ITmD37E was discovered in a wide range of mosquito species. Sequence analysis of multiple copies in three Aedes species showed similar terminal inverted repeats and common putative TA target site duplications. The ITmD37E transposases contain a conserved DD37E catalytic motif, which is unique among reported transposons of the IS630-Tc1-mariner superfamily. Sequence comparisons and phylogenetic analyses suggest that ITmD37E forms a novel family distinct from the widely distributed Tc1 (DD34E), mariner (DD34D), and pogo (DDxD) families in the IS630-Tc1-mariner superfamily. The inclusion in the phylogenetic analysis of recently reported transposons and transposons uncovered in our database survey provided revisions to previous classifications and identified two additional families, ITmD37D and ITmD39D, which contain DD37D and DD39D motifs, respectively. The above expansion and reorganization may open the doors to the discovery of related transposons in a broad range of organisms and help illustrate the evolution and structure-function relationships among these distinct transposases in the IS630-Tc1-mariner superfamily. The presence of intact open reading frames and highly similar copies in some of the newly characterized transposons suggests recent transposition. Studies of these novel families may add to the limited repertoire of transgenesis and mutagenesis tools for a wide range of organisms, including the medically important mosquitoes.


2006 ◽  
Vol 61 (5-6) ◽  
pp. 341-346 ◽  
Author(s):  
Jae-Woo Park ◽  
A. M. Abd El-Aty ◽  
Myoung-Heon Lee ◽  
Sung-Ok Song ◽  
Jae-Han Shim

A multiresidue method for the simultaneous determination of 22 organochlorine (OCs) and organophosphorus (Ops) pesticides (including isomers and metabolites), representing a wide range of physicochemical properties, was developed in fatty matrices extracted from meat. Pesticides were extracted from samples with acetonitrile/n-hexane (v :v, 1:1). The analytical screening was performed by gas chromatography coupled with electron-capture detection (ECD). The identification of compounds was based on their retention time and on comparison of the primary and secondary ions. The optimized method was validated by determining accuracy (recovery percentages), precision (repeatability and reproducibility), and sensitivity (detection and quantitation limits) from analyses of samples fortified at 38 to 300 ng/g levels. Correlation coefficients for the 22 extracted pesticide standard curves (linear regression analysis, n = 3) ranged from 0.998 to 1.000. Recovery studies from 2 g samples fortified at 3 levels demonstrated that the GC-ECD method provides 64.4-96.0% recovery for all pesticides except 2,4′-DDE (44.6-50.4%), 4,4′-DDE (51.1-57.5%) and 2,4′-DDT (50.0-51.2%). Both repeatability and reproducibility relative standard deviation values were < 20% for all residues. Detection limits ranged from 0.31 to 1.27 ng/g and quantification limits were between 1.04 and 4.25 ng/g. The proposed analytical method may be used as a simple procedure in routine determinations of OCs and Ops in meat. It can also be applied to the determination of pesticide multi-residues in other animal products such as butter and milk.


mBio ◽  
2018 ◽  
Vol 9 (6) ◽  
Author(s):  
Christoph M. Ernst ◽  
Christoph J. Slavetinsky ◽  
Sebastian Kuhn ◽  
Janna N. Hauser ◽  
Mulugeta Nega ◽  
...  

ABSTRACTDaptomycin, a calcium-dependent lipopeptide antibiotic whose full mode of action is still not entirely understood, has become a standard-of-care agent for treating methicillin-resistantStaphylococcus aureus(MRSA) infections. Daptomycin-resistant (DAP-R)S. aureusmutants emerge during therapy, featuring isolates which in most cases possess point mutations in themprFgene. MprF is a bifunctional bacterial resistance protein that synthesizes the positively charged lipid lysyl-phosphatidylglycerol (LysPG) and translocates it subsequently from the inner membrane leaflet to the outer membrane leaflet. This process leads to increased positiveS. aureussurface charge and reduces susceptibility to cationic antimicrobial peptides and cationic antibiotics. We characterized the most commonly reported MprF mutations in DAP-RS. aureusstrains in a defined genetic background and found that only certain mutations, including the frequently reported T345A single nucleotide polymorphism (SNP), can reproducibly cause daptomycin resistance. Surprisingly, T345A did not alter LysPG synthesis, LysPG translocation, or theS. aureuscell surface charge. MprF-mediated DAP-R relied on a functional flippase domain and was restricted to daptomycin and a related cyclic lipopeptide antibiotic, friulimicin B, suggesting that the mutations modulate specific interactions with these two antibiotics. Notably, the T345A mutation led to weakened intramolecular domain interactions of MprF, suggesting that daptomycin and friulimicin resistance-conferring mutations may alter the substrate range of the MprF flippase to directly translocate these lipopeptide antibiotics or other membrane components with crucial roles in the activity of these antimicrobials. Our study points to a new mechanism used byS. aureusto resist calcium-dependent lipopeptide antibiotics and increases our understanding of the bacterial phospholipid flippase MprF.IMPORTANCEEver since daptomycin was introduced to the clinic, daptomycin-resistant isolates have been reported. In most cases, the resistant isolates harbor point mutations in MprF, which produces and flips the positively charged phospholipid LysPG. This has led to the assumption that the resistance mechanism relies on the overproduction of LysPG, given that increased LysPG production may lead to increased electrostatic repulsion of positively charged antimicrobial compounds, including daptomycin. Here we show that the resistance mechanism is highly specific and relies on a different process that involves a functional MprF flippase, suggesting that the resistance-conferring mutations may enable the flippase to accommodate daptomycin or an unknown component that is crucial for its activity. Our report provides a new perspective on the mechanism of resistance to a major antibiotic.


Radiocarbon ◽  
2012 ◽  
Vol 54 (3-4) ◽  
pp. 879-886 ◽  
Author(s):  
Fiona Brock ◽  
Rachel Wood ◽  
Thomas F G Higham ◽  
Peter Ditchfield ◽  
Alex Bayliss ◽  
...  

A recent study into prescreening techniques to identify bones suitable for radiocarbon dating from sites known for poor or variable preservation (Brock et al. 2007, 2010a) found that the percent nitrogen (%N) content of whole bone powder was the most reliable indicator of collagen preservation. Measurement of %N is rapid, requires little preparation or material, and is relatively cheap. The technique reduces the risk of needlessly sampling valuable archaeological objects, as well as saving time and money on their unsuccessful pretreatment prior to dating. This method of prescreening is now regularly used at the Oxford Radiocarbon Accelerator Unit (ORAU). In the original study, linear regression analysis of data from 100 bones from 12 Holocene sites across southern England showed that when 0.76% N was chosen as a threshold, 84% of bones were successfully identified as containing sufficient (i.e. >1%) or insufficient (i.e. <1%) collagen for dating. However, it has been observed that for older, Pleistocene bones the failure rate may be higher, possibly due to the presence of more degraded, short-chain proteins that pass through the ultrafilters used in pretreatment, resulting in lower yields. Here, we present linear regression analysis of data from nearly 600 human and animal bones, antlers, and teeth, from a wide range of contexts and ages, to determine whether the 0.76% threshold identified in the previous study is still applicable. The potential of carbon:nitrogen atomic weight ratios (C:N) of whole bone to predict collagen preservation is also discussed.


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