Real-time PCR Tests in Dutch Exotic Mosquito Surveys; Implementation of Aedes aegypti and Aedes albopictus Identification Tests, and the Development of Tests for the Identification of Aedes atropalpus and Aedes japonicus japonicus (Diptera: Culicidae)

2015 ◽  
Vol 52 (3) ◽  
pp. 336-350 ◽  
Author(s):  
B. T. L. H. van de Vossenberg ◽  
A. Ibanez-Justicia ◽  
E. Metz-Verschure ◽  
E. J. van Veen ◽  
M. L. Bruil-Dieters ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Real Time ◽  
Aedes Albopictus ◽  
Real Time Pcr ◽  
Aedes Japonicus ◽  
Aedes Japonicus Japonicus ◽  
Aedes Atropalpus

scholarly journals Implementation of a deep learning model for automated classification of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) in real time

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Song-Quan Ong ◽  
Hamdan Ahmad ◽  
Gomesh Nair ◽  
Pradeep Isawasan ◽  
Abdul Hafiz Ab Majid
Keyword(s):  
Deep Learning ◽  
Aedes Aegypti ◽  
Real Time ◽  
Aedes Albopictus ◽  
Automated Classification ◽  
Expert Performance ◽  
Deep Convolutional Neural Networks ◽  
Significant Difference ◽  
Set Up

AbstractClassification of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) by humans remains challenging. We proposed a highly accessible method to develop a deep learning (DL) model and implement the model for mosquito image classification by using hardware that could regulate the development process. In particular, we constructed a dataset with 4120 images of Aedes mosquitoes that were older than 12 days old and had common morphological features that disappeared, and we illustrated how to set up supervised deep convolutional neural networks (DCNNs) with hyperparameter adjustment. The model application was first conducted by deploying the model externally in real time on three different generations of mosquitoes, and the accuracy was compared with human expert performance. Our results showed that both the learning rate and epochs significantly affected the accuracy, and the best-performing hyperparameters achieved an accuracy of more than 98% at classifying mosquitoes, which showed no significant difference from human-level performance. We demonstrated the feasibility of the method to construct a model with the DCNN when deployed externally on mosquitoes in real time.

Differences in Responses to Urbanization Between Invasive Mosquitoes, Aedes japonicus japonicus (Diptera: Culicidae) and Aedes albopictus, in Their Native Range, Japan

2019 ◽  
Vol 57 (1) ◽  
pp. 104-112 ◽  
Author(s):  
Motoyoshi Mogi ◽  
Peter A Armbruster ◽  
Nobuko Tuno
Keyword(s):  
Aedes Albopictus ◽  
Blood Feeding ◽  
Forest Tree ◽  
Careful Examination ◽  
Late 20Th Century ◽  
Aedes Japonicus ◽  
Domestic Species ◽  
Aedes Japonicus Japonicus ◽  
Distribution Ranges ◽  
Tree Holes

Abstract The Asian mosquitoes, Aedes albopictus (Skuse) and Aedes japonicus japonicus (Theobald), have invaded North America, Europe, and other regions since the late 20th century. In invasive ranges, Ae. albopictus has well been recognized as urban, domestic species, whereas views about the macrohabitats of Ae. j. japonicus are inconsistent. Previous reports in Japan suggest the disappearance of Ae. j. japonicus from metropolises. However, container-mosquito larvae have not been inspected simultaneously for various macro and microhabitats in metropolises in Japan. The current study in Fukuoka City, a metropolis in southwest Japan, confirmed the absence of Ae. j. japonicus irrespective of macrohabitats (temples with graveyards, shrines, public graveyards, cultivated bamboo groves, and urban forests, all within the area densely inhabited by humans) and microhabitats (container types). In contrast, Ae. albopictus was dominant throughout the macro and microhabitats except forest tree holes rich with competitive species. Past records indicate the disappearance of Ae. j. japonicus from metropolitan Fukuoka within the last 70 yr. Based on careful examination of available evidence, we concluded that 1) both species benefit from human-made environments with artificial containers free from competitors, 2) Ae. j. japonicus disappeared due to hot, dry summer conditions facilitated by urban heat-island effects and a decrease in favored mammal hosts, and 3) Ae. albopictus has proliferated with higher tolerance to hot, dry climate and a wider blood-feeding host-range including humans. This difference is important for efficient control of each species as well as predicting and preventing the expansion into new distribution ranges.

scholarly journals Diagnostic primers to distinguish between wMel and wAlbB Wolbachia infection - A simplified approach for use in field-collected Aedes aegypti in dengue reduction trials

2021 ◽  
Author(s):  
Meng-Jia Lau ◽  
Ary A. Hoffmann ◽  
Nancy Endersby-Harshman
Keyword(s):  
Aedes Aegypti ◽  
Relative Density ◽  
Real Time ◽  
Real Time Pcr ◽  
Wolbachia Infection ◽  
Simplified Approach ◽  
Efficient Estimate ◽  
Pcr Assays

Detection of the Wolbachia endosymbiont in Aedes aegypti mosquitoes through real-time PCR assays is widely used during and after Wolbachia releases in dengue reduction trials involving the wMel and wAlbB strains. However, primers applied in current successful Wolbachia releases cannot be used in a single assay to distinguish between these strains. Here, we developed a new diagnostic primer pair wMA which can detect wMel or wAlbB infection in the same assay. We also tested current Wolbachia primers and show that there is variation in their performance when they are used to assess the relative density of Wolbachia. The new wMA primers provide an accurate and efficient estimate of the presence and density of both Wolbachia infections.

scholarly journals Optimasi Analisis Melting Curve untuk Skrining Cepat dan Sensitif Mutasi V1016G pada Aedes aegypti Resisten Sintetik Piretroid dengan Reaksi Rantai Polimerase Spesifik Alel

2021 ◽  
pp. 153-160
Author(s):  
Dyah Widiastuti ◽  
Agustiningsih Agustiningsih ◽  
Ihda Zuyina Ratna Sari ◽  
Tri Ramadhani
Keyword(s):  
Aedes Aegypti ◽  
Real Time ◽  
Melting Temperature ◽  
Real Time Pcr ◽  
Pyrethroid Resistance ◽  
Melting Curve ◽  
Previous Method ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Wild Type

Detection of V1016G mutation is important for identifying the mechanism of  synthetic pyrethroid resistance in Aedes aegypti population. The previous method has described an allele specific polymerase chain reaction (AS-PCR) using conventional PCR to detect the mutation. Although the method has great differentiating power and reproducibility, faster and more sensitive genotyping method is essential to accurately detect the mutation. This study evaluate the used of SYBR® Green real-time PCR and melting curve analysis (MCA) to identify the V1016G mutation. The collection of homozygous 1016G, heterozygous, and wild type (1016 V) mosquitoes DNA genome was extracted using genomic DNA mini kit. The SsoAdvanced™ Universal SYBR® Green Supermix was used to identify alleles by real-time PCR followed melting curve analysis of the amplicons. Melting curve analysis produced reproducible results for the loci tested. The melting temperature was reached at 78.5 oC for homozygous 1016G mosquito and at 86 oC for wild type mosquito. Meanwhile, the heterozigous mosquito revealed two peaks of melting temperature at both 78.5 oC and 86 oC. These easily interpretable and distinguishable melting curve results were consistent with AS-PCR results obtained for the same alleles. The described MCA application for screening V1016G mutation is fast and widely accessible also could be implemented under field conditions

scholarly journals Field-deployable molecular diagnostic platform for arbovirus detection in Aedes aegypti

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Natalie Rutkowski ◽  
Yuemei Dong ◽  
George Dimopoulos
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Diagnostic System ◽  
Molecular Diagnostic ◽  
Infection Prevalence ◽  
The Novel ◽  
Genomic Rna ◽  
Viral Genomic Rna

Abstract Background Surveillance of mosquito infection status is critical for planning and deployment of proper mosquito control initiatives. Point-of-care (POC) detection assays are necessary for monitoring the infection prevalence and geographical range of viruses in mosquito vector populations. We therefore assessed the novel real-time PCR (qPCR) bCUBE (Hyris, London, UK) molecular diagnostic system as a tool for virus detection. Methods Aedes aegypti Rps17 was used to validate and determine correlation coefficient for the novel bCUBE qPCR system to a laboratory standard StepOnePlus real-time PCR system (Applied Biosystems, Waltham, MA, USA). Experimentally infected Ae. aegypti were quantified for Zika (ZIKV) and dengue virus serotype 2 (DENV2) viral genomic RNA. Infection prevalence was compared to plaque assay. Results We developed and validated a novel qPCR system for the detection of ZIKV and DENV2 using the real-time qPCR system bCUBE. With bCUBE-based qRT-PCR, viral genomic RNA could be detected in individually infected Ae. aegypti mosquitoes and in pools of 5, 10 or 15 mosquitoes. Conclusions The portable qPCR bCUBE diagnostic system is capable of detecting Zika and dengue virus in mosquitoes and therefore has potential as a practical field-deployable diagnostic test for vector-borne disease surveillance programmes.

scholarly journals Evaluation of reference genes at different developmental stages for quantitative real-time PCR in Aedes aegypti

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Najat Dzaki ◽  
Karima N. Ramli ◽  
Azali Azlan ◽  
Intan H. Ishak ◽  
Ghows Azzam
Keyword(s):  
Aedes Aegypti ◽  
Real Time ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Developmental Stages ◽  
Quantitative Real Time Pcr

Aedes Vector Surveillance in the Southeastern United States Reveals Growing Threat of Aedes japonicus japonicus (Diptera: Culicidae) and Aedes albopictus

2019 ◽  
Vol 56 (6) ◽  
pp. 1745-1749
Author(s):  
Benjamin A McKenzie ◽  
Kelly Stevens ◽  
Ansley E McKenzie ◽  
Jovana Bozic ◽  
Derrick Mathias ◽  
...  
Keyword(s):  
United States ◽  
Aedes Albopictus ◽  
Gulf Coast ◽  
The United States ◽  
Time Frame ◽  
The State ◽  
Third Wave ◽  
Vector Surveillance ◽  
Aedes Japonicus ◽  
Aedes Japonicus Japonicus

Abstract Mosquito-borne diseases are on the rise globally, and have the potential to thrive along the Gulf Coast of the United States, where subtropical conditions may facilitate the introduction or movement of mosquito vectors. Despite surveillance efforts, Aedes aegypti (L.) had not been detected in the Gulf state of Alabama for nearly three decades. The detection of Ae. aegypti in Alabama may suggest remnant or reemergent populations of this vector. We conducted adult sampling between May and August of 2018 to capture mosquitoes during a time frame when all species should be active. This was to ensure no species were missed due to overwintering and to identify the distributions of Aedes mosquitoes of medical importance. No Ae. aegypti were detected in Alabama over the period of this study. We detected Aedes albopictus (Skuse) in 65 counties and the recently invasive Aedes japonicus japonicus (Theobald) in 30 counties across the state. These results indicate that while Ae. aegypti was recently reported from parts of Alabama, the state is not experiencing a major resurgence of the species, whereas Ae. albopictus remains ubiquitous. Further, results indicate that a third wave of Aedes invasion may be occurring, that of Ae. japonicus japonicus. All three of these species are medically important vectors and may pose threats to the public health of the Gulf Coast of the United States.

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