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attraction response of mosquitoes when exposed to CDC light traps

Master’s degree proposal
Aedes aegypti
As a well-known vector of diseases such as Yellow Fever, Dengue, Chikungunya, and Zika virus the Aedes aegypti have remained a global health issue. The ancestral roots of this species are suspected to have begun in the sub-Saharan region on the continent of Africa as a tree-hole laying mosquito that feeds primarily on warm-blooded animals (Brown et al. 2011). Since its rapid spread around the world, this species of Aedes has been successful in spreading to most of the warmer regions along the equator and is now found in large areas of Asia and most of North and South America (Lambrechts et al. 2011). With a well-documented expansion amongst urbanized human settlements, the number of illnesses transferred by this species has only increased with population growth and expansion over the last 50 years (Weaver 2014). Artificial human-made containers that store water, such as used tires, provide perfect breeding sites for Aedes aegypti and has resulted in these mosquitoes being shipped around the world through the movement of these containers (Harrington, et al. 2001). Over time, the preference of host feeding has changed in this species from their original zoophilic tendencies to the modern-day anthropophilic feeding behavior with adjusted feeding times primarily to the period when humans are most active. (Scott and Takken 2012). Aedes aegypti feeding times are reported by the Center for Disease Control and Prevention (CDC) as mostly occurring several hours prior to sunset and for two hours after the sun rises (CDC.GOV 2012). These adaptions to diurnal biting habits and being most active during the day likely contributes to the mosquitoes’ showing little attraction to the incandescent light sources used in standard suction traps (Hoel, et al. 2009; Thurman and Thurman 1955).
Centers for Disease Control and Prevention Light Trap (CDC light trap)
The first initial creation of the battery-operated CDC light trap in the 1960’s by Drs. Dan Sudia and Roy Chamberlain provided a lighter alternative to earlier suction light trap designs (Sudia and Chamberlain 1962). Since its creation, the CDC light trap has become one of, if not the most widely used trapping systems for insect surveillance programs (Cohnstaedt et al. 2008; McDermott and Mullens 2017). After shortcomings of incandescent light bulbs were identified, the utilization of UV bulbs began in the 1950’s and significantly increased the capture rate of light traps (Pfrimmer 1955). In attempts to improve the capture yield of these traps, several additions to surveillance traps have been implemented over time. A well-known attractant used by many entomologists, which bolstered the effectiveness of these traps, is CO2 dispensed from a canister or produced by dry ice (Reeves 1951). In addition to CO2, many chemical attractants have been created in the attempt to increase the capture rate of light traps; these include 1-octen-3-ol, lactic acid, butanone, and BG Lure with various success (Kline et al. 1994; Kline and Mann 1998; Dekker et al. 2002). A more recent innovation in light trap technology is the use of a wide range of different colored LED’s to save energy and attempt to improve capture counts (Hoel et al. 2007; Cohnstaedt et al. 2008).
Optical Attraction in Aedes aegypti
Studies have determined that mosquitoes are attracted to 365nm wavelength of light when used in surveillance traps (Burkett and Butler 2005). The overall lack of attraction to the incandescent light utilized in many mosquito traps is thought to be due to mosquitoes primarily seeing in the ultraviolet, blue, and green spectra of color (Briscoe and Chittka 2001). Incandescent bulbs cover a wide area of the light spectrum with most light in red/infrared wavelengths and only minor presence of the lower blue, green, and the ultraviolet ranges of light (Cohnstaedt et al. 2008). UV light is described as wavelengths less than 400nm and exposure to this light source can influence behaviors such as mating, host-seeking, circadian rhythms, and oviposition, through mediation within their compound eye (Day 2005; Hunt et al. 2001). The compound eyes found in mosquitoes follow a typical arrangement of the retinal photoreceptors R1 through R8 configuration, each rhabdomere expressing various rhodopsin’s (Brammer 1970, Hu et al. 2009). Testing of photoreceptors and rhodopsin genes expression in Aedes aegypti through exposure to ultraviolet light has shown that these insects’ optical genes are responsive to distinct UV spectra (Hu et al., 2014).
Advances in Light Sources.
Most of the suction-based light traps have utilized incandescent or UV fluorescent bulbs as attractants until more recent additions of LED technologies have been implemented (McDermott and Mullens 2017). LED can produce a very narrow range of bandwidth to entice the preferred insect. These ranges can be set between 350nm (UV) and 700nm (infrared) with the highest quality LEDs showing an omnidirectional spread of light to 120-degree angle (Cohnstaedt et al. 2008). These refined uses of specific spectra of UV light spectrum could prove pivotal in attracting Aedes aegypti, and other desired species because many insects can detect changes of wavelength as minute as 10 nm (Snyder et al. 2016). During a test of a UV black light (BL) bulb and a UV LED in a Malaria-endemic area of South Korea, significantly larger capture rates and diversity of species captured were obtained with UV LED lights with a peak at the 365 nm wavelength (Kim et al. 2017). Among the species caught in the test conducted in South Korea, the most frequently captured mosquito was identified as Aedes vexans which accounted for over 80% of the total capture in both the BL and UV LED versions of the trap (Kim et al. 2017). The upgrades in quality of LED lights, reduction in price, and utilization of specific light spectra provide the potential for increasing the effectiveness of light traps.
Compare the attraction response of mosquitoes when exposed to CDC light traps equipped with incandescent bulbs, black-light fluorescent bulbs and UV LED Lights.
– Compare the individual capture rates over time ofAedes aegypti males, host-seeking females, and blood-fed gravid females in a laboratory setting.
– Evaluate the trap preference of Aedes aegypti males, host-seeking females, and blood-fed gravid females when traps are run against one another in an enclosed laboratory environment.
– Compare the capture of native mosquitoes’ species in field locations
We hypothesize that the modification of the CDC light trap utilizing the SVC UV LED modified light source will be more attractive to lab reared Aedes aegypti and various outdoor species than CDC light traps utilizing BL and incandescent bulbs.
Scope of Research
With the prices of LED technology dropping and the increase in the quality of UV LED’s coming onto the market, further testing and evaluation of both quality and effectiveness of these new lights are needed. These tests will focus on the attractiveness of a CDC light trap modified with UV LED when compared with the CDC light trap equipped with BL bulb or incandescent bulb. This trap and others utilizing LED technologies could improve vector surveillance yields, decrease trap energy requirements, and lower cost of surveillance when compared with current trapping methods.
Materials and Methods
Aedes aegypti will be provided from the “Orlando” strain colonies maintained at the United States Department of Agriculture (USDA-ARS) located in Gainesville, Florida. Mosquitoes will be used between 1 and 3 days old for males, and 4 to 7 days old for females to ensure full visual development and mating. During this development time, the specimens will be provided with full access to 10% sugar water for feeding. Ageing of the mosquitoes will be accomplished by allowing pupae a 24-emergence time before moving the pupae into a new unoccupied cage and repeating the process. The bioassays of males will be conducted on days 1 to 3 after emergence, followed by the females from days 4 to 7. When blood feeding is required for females in experiments, it will be from chickens maintained by Urban Entomology staff following (IACUC 201603836) protocols. Blood feeding will happen on day four of female mosquito development with testing being run 48 hours after blood feeding on day 7. Specimens will be aspirated and chilled briefly in a lab freezer and placed on ice before sorting, counting, and putting into groups for release into enclosures.
Light traps
Lights will be examined via an Ocean Optics® 2000 USB spectrometer.
– Trap (A) – CDC Trap modified to utilize the MOSCLEAN UV LED component
SVC 6 LED CMTB_LED_160414_REV01 Obtained from Soul Viosys company, Soul South Korea.

– Trap (B) – CDC Trap with BL fluorescent tube.
John W. Hock model number 812 Ordered from John W. Hock company Gainesville FL.

Bulb F4T5/BLB 130V 4W Ordered from John W. Hock company Gainesville FL.

– Trap (C) – CDC trap with a standard incandescent wing nut light bulb
John W. Hock model number 512 Ordered from John W. Hock company Gainesville FL.

Eiko Certified Green bulb number 47 bulb Purchased from Batteries Plus Bulbs Gainesville FL

Indoor trap capture rate
Timed capture-rate tests will be conducted in a darkened room with three separate testing groups of Aedes aegypti males, host-seeking, and blood fed gravid females. Each group of 50 mosquitoes will be released into separate 24X24x24 inch rearing cages containing either a CDC with UV LED, CDC with UV BL, or CDC with incandescent lights. Trap location within the room will be rotated after each repetition. Mosquitoes will be given 30 minutes to wake up after being sedated over ice in small containers and then an additional 15 minutes within the rearing enclosure to acclimate to the environment before starting the traps. The room will be kept at 23 and 26 degrees Celsius and overhead room lighting will be turned off to minimize competing light sources in between counts. After the traps have been launched, capture counts within each will be hourly over 4 hours. This process will be repeated for male, host-seeking females, and 48-hour gravid females for a total of 10 repetitions.
Indoor trap preference
Three trap preference testing will be compared in an enclosed room utilizing trap A, B, and C. Groups of 100 mosquitoes will be released into a controlled room with each trap placed in a separate corner of the room and rotated after each run. Mosquitoes will be allotted 30 minutes to wake up after being sedated over ice in a small container and then released into the room with the overhead lighting turned off to minimize any competing light sources. The testing will start between 4:00 and 5:00 each afternoon and will be stopped and collected in black fine meshed draw string bags between 9:00 and 10:00 each morning. This process will be repeated for male, host-seeking females, and 48-hour gravid females for a total of 10 repetitions.
Outdoor trap catches
An outdoor traps comparison run utilizing (A), (B), and (C). Three locations where selected in UF’s NATL and the surrounding area. Site selection will be limited to areas in which power sources can be utilized to run the UV LED light source attachment. Sites will be labeled as one, two, and three with individual traps being placed at each one. After each rep, the traps will be rotated to the next site. A total of 30 nights of trapping will be conducted starting between 4:00 to 5:00 in the afternoon and completed between 9:00 and 10:00 in the morning. The number captured in each trap will be documented and mosquitoes will be identified down to species and compiled.
Preliminary Statistical Analysis
Preliminary Results
Indoor trap capture rate

Indoor trap preference

Outdoor trap catches

Research Schedule
Spring 2018
– Three trap capture rate tests
Blood fed gravid females 48 hour
– Three trap preference room testing
Blood fed gravid females 48 hour
Summer 2018
– Schedule exit seminar and thesis defense
– Schedule graduation
Adriana D. Briscoe and Lars Chittka. 2001.The Evolution of Color Vision in Insects Annual Review of Entomology 2001 46:1, 471-510
Brammer J. D. 1970. The ultrastructure of the compound eye of a mosquito Aedes aegypti L. J. Exp. Zool. 175, 181-195
Brown, J. E., C. S. Mcbride, P. Johnson, S. Ritchie, C. Paupy, H. Bossin, J. Lutomiah, I. Fernandez-Salas, A. Ponlawat, A. J. Cornel, W. C. Black, N. Gorrochotegui-Escalante, L. Urdaneta-Marquez, M. Sylla, M. Slotman, K. O. Murray, C. Walker, and J. R. Powell. 2011. Worldwide patterns of genetic differentiation imply multiple domestications of Aedes aegypti, a major vector of human diseases. Proceedings of the Royal Society B: Biological Sciences. 278: 2446–2454.
Burkett, D. A., and J. F. Butler. 2005. Laboratory Evaluation of Colored Light As An Attractant For Female Aedes Aegypti, Aedes Albopictus, Anopheles Quadrimaculatus, And Culex Nigripalpus. Florida Entomologist. 88: 383–389. (2012). [online] Available at: [Accessed 5 Nov. 2018].
Cohnstaedt, L. W., J. I. Gillen, and L. E. Munstermann. 2008. Light-Emitting Diode Technology Improves Insect Trapping. Journal of the American Mosquito Control Association. 24: 331–334.
Day, J.F. 2005. Host-seeking strategies of mosquito disease vectors. Journal of the American Mosquito Control Association 21: 17–22.
Emily G McDermott, and Bradley A Mullens 2017. The Dark Side of Light Traps, Journal of Medical Entomology, Volume 55, Issue 2, 28 February 2018, Pages 251–261
Harrington, L. C., J. D. Edman, and T. W. Scott. 2001. Why Do Female Aedes aegypti (Diptera: Culicidae) Feed Preferentially and Frequently on Human Blood? Journal of Medical Entomology. 38: 411–422.
Hoel, D. F., Kline, D. L.,

Cerebral Infarction of the Uygur and Han Ethnic Groups

Research Article
The Difference of the Polymorphism of RS4360791 Loci of ALOX5AP Gene between Cerebral Infarction of the Uygur andHan Ethnic Groups InXinjiang
1. Abstract Objective:To explore the difference of the polymorphism of RS4360791 loci of ALOX5AP gene between cerebral infarction of the Uygur and Han ethnic groups in Xinjiang Province, China.
Methods:This is a case-control study. The polymorphism of the genotype of RS4360791 loci of ALOX5AP gene and the frequency of its allele were tested using polymerase chain reaction and DNA sequencing. And the association between different genotypes and the risks of cerebral infarction of Uygur and Han ethnic groups were analyzed using Pearson chi square test and multivariate logistic regression model.
Results: There was no significant difference in the ALOX5AP gene rs4360791 between the Uygur and the Han (P>0.05).
Conclusion: In this study, the ALOX5AP gene RS4360791 polymorphisms in Xinjiang Uygur and Han nationality cerebral infarction were studied: this gene positions between cerebral infarction in Han and Uyghur The point genotype and gene frequency were not statistically significant (P>0.05).
Key words: 5-lipoxygenase activator protein; Ischemic stroke; Gene; Atherosclerosis
2. Introduction Ischemic stroke is a complicated disease related with different factors and its prevalence increases with risk factors like hypertension, diabetes and smoking. A considerable number of ischemic stroke attacks are caused by vessel stenosis and emboli derived from unstable plaques. Arterial atherosclerosis is one the major pathology processes of ischemic stroke and inflammatory factors are key elements to trigger atherosclerosis. Analysis of inflammatory factors can help understand the risk factors of ischemic stroke, assisting treatment and second prevention of stroke and lowering the burden of families and societies.
Arterial atherosclerosis is a chronic inflammatory disease, contributing to cerebrovascular diseases [1-2]. Leukotriene, the product of the 5- lipoxygenase activator protein (5-LAP) and 5-lipoxygenase(5-LO) pathway, accelerates the process of atherosclerosis. It promotes lipid streak formation, stable/unstable plaque pathological process and plaque rupture, resulting in cerebrovascular diseases.
ALOX5AP related mutation may be associated with excretion of substances like leukotriene, leading to arterial atherosclerosis formation, and subsequently vessel stenosis, and plaque rupture as well as the final the ischemic stroke caused by arterial vessel occlusion [3]. The aim of this study is to explore the relationship between the polymorphism RS4360791 loci of ALOX5AP gene and artery atherosclerosis

related stroke, investigating the difference of the polymorphism of RS4360791 loci between cerebral infarction of the Uygur and Han ethnic groups.
3. Methods
Patient selection
Patients diagnosed with cerebral infarction from September 2017 to December 2018 in the Neurology Department of Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous region were enrolled in this study. A total number of 100 patients were included, 50 Han patients and 50 Uygur patients. Every patient underwent complete computer tomography (CT) and magnetic resonance imaging (MRI) scan and was clinically evaluated and diagnosed by at least two attending doctors. And
patients were divided into 2 groups:
(1) Case group: 50 Uygur cerebral infarction patients, including 32 males and 18 females. Age range from 54-76 years with average (SD) 66.42 (0.99) years. 2). Control group: 50 Han cerebral infarction patients, including 27 males and 23 females. Age range from 50-70 years with average (SD) 61.54 (0.71) years. Informed consent was obtained from each patient. And this study was approved by ethic committees of Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous region.
Baseline demographic and clinical data collection
Every participant was registered after signing the informed consent. Baseline demographic and clinical data including sex, age, ethic group, history of smoking, history of alcohol drinking, occupation, etc. History of smoking were defined as more than 10 cigarettes per day for at least 2 years.
Blood sample collection
Blood sample collection and AOLX5AP gene assay
For every participant enrolled, 5mL of peripheral blood using Ethylene Diamine Tetra-acetic Acid (EDTA) tube was collected at the first morning after admission when fasting, Then the blood samples were stored in a fridge of -80℃after primary centrifugation. And direct sequencing was used to verify with the Kompetitive Allele Specific (KASP) genotyping.
Statistical Analysis
Statistical analysis was performed using SPSS 18.0 (IBM, New York). Categorical variables were analyzed using chi square test and continuous variables were analyzed using t test. The Hardy-Weinberg equation was tested using chi square goodness of fit test. Pearson chi square test was used to analyze the difference of genotype and alleles between the case group and the control group.
4. Results
Comparison of baseline data between groups
First the baseline data including demographic and clinical data were analyzed between the case group and the control group. (Table 1) No statistical significance between the two groups with respect to sex, age, history of hypertension, history of diabetes mellitus and history of smoking (P>0.05) was discovered except for history of hyperlipidemia(p<0.05).
Table 1: Comparison of general clinical data between case group and control group (%, x  s )

Baseline Data
Case Group (n=50)
Control Group
(n 50)
χ2 Value or
t Value
of Smoking[n (%)]
History of Hypertension[n (%)]
History of Hyperlipidemia [n (%)]
History of Diabetes Mellitus[n (%)]
Results of total DNA extraction
Total DNA of the studies samples was extracted and underwent quality control. All the samples were qualified to undergo subsequent experiments. The electrophoresis of total DNA extraction is seen in Figure 1.

Figure 1: Electrophoresis of total DNA extraction.
Markers from top to bottom were 100bp, 250bp, 500bp, 750bp, 1000bp, 2000bp. A qualified band of a genotype should be a single clear and bright band above 2000bp.
Results of KASP genotyping
3 loci of rs3369790 could be clearly distinguished using KASP genotyping. (Figure 2)

Figure 2: SNP locus mapping.
Results of DNA direct sequencing
In order to further verify the SNP loci, samples were randomly chosen to undergo direct sequencing to verify the results of KASP genotyping. First the of all, the DNA samples were amplificated using PCR and then direct sequencing was performed to obtain corresponding sequences. The sequence diagram of each site can be seen in Figure 2.
Comparison of the polymorphism of the RS4360791 loci of ALOX5AP between Uygur and Han Groups
Through the sequencing of the polymorphism RS436071 loci of the case group (Uygur patients) and control group (Han patients), 3 types of genotype, GG, AG. AA were discovered. The analysis of the frequency of 3 genotypes and A/G distribution demonstrated that there was no statistical significance
for the distribution of 3 genotypes between case and control group (χ2=1.190, P=0.275>0.05) and the
overall distribution of alleles A/G also showed no statistical difference (χ2=0.1.087, P=0.297>0.05).
(Table 2)
Table 2: Comparison of genotype and allele frequencies of rs4360791 locus between case group and control group [n(%)].
Genotype Frequency
Allele Frequency
Case group
Control group
The pathophysiology of artery atherosclerosis related stroke could be mainly described as a sudden decrease or blockage of supplying arterial blood flow, leading to ischemia and hypoxia of the brain tissue and the subsequently brain infarction. This pathological process was accompanied by

corresponding clinical symptoms and syndromes of neurological deficits, such as hemiparalysis, aphasia, etc. [4] Cerebral infarction is a common neurological disease with complicated pathologies. It is globally acknowledged that the risk factors of cerebral infarction include hyperlipidemia, hypertension, history of smoking, history of alcohol drinking, history of diabetes mellitus, obesity and family history of stroke. And with the development of molecular biology, the association between cerebral infarction and gene polymorphism has been drawing increasing attention. [5]
Previous studies have demonstrated that inflammation is the key element that contributes to the pathogenesis of atherosclerosis.
5- lipoxygenase activator protein, encoded by ALOX5AP gene, is the crucial regulator of the biosynthesis of leukotriene (LTS), leading to fat deposits in the arteries. LTS elicits white blood cell activation and monocyte adhesion into the vessel wall, which plays an important role in atherosclerosis and inflammatory disorders. ALOX5AP is strongly associated with the formation and development of atherosclerosis and plaques, the prevalence of which is further raised by ALOX5AP mutation. [6]
ALOX5AP is key to the inflammatory process, such as the production of leukotriene. [7] The bioactive products of certain loci of ALOX5AP are related to many cardiocerebrovascular diseases. [8-12] Additionally, many clinical trials of Western countries suggested that ALOX5AP took part in the pathophysiological process of atherosclerosis, resulting in ischemic strokes. And recent studies have shown some certain loci of ALOX5AP was related with atherosclerosis in Chinese population. This study adapted a case-control design where genotypes of polymorphism of RS4360791 loci of ALOX5AP and allele frequencies of 100 patients were studies using PCR and DNA sequencing and the association between different genotypes and risks of cerebral infarction was compared between Uygur and Han ethic groups. Results of this study showed that there was no significant difference between 2 groups with respect to sex, age, history of smoking, history of hypertension and history of diabetes. The only statistical difference between 2 groups was history of hyperlipidemia, which could be accounted for the different eating habits of these 2 ethic groups. The Uygur, one of the national minorities of China, are fond of meat, diary, fried food and high sugar food and fruits rather than high fiber food such as vegetables. Additionally, dinners were of great importance in their daily life. Therefore, the Uygur manifest with hyperlipidemia and high body weight index. Contrarily, the Han’s diet contains much less calorie. Therefore, this study can help control relative risk factors with statistical support.
Through the DNA sequencing of the polymorphism of RS4360791 loci of the case group (the Uygur) and the control group (the Han), 3 types of genotypes were discovered. And difference of frequency of these 3 genotypes and C/G alleles between 2 groups was further analyzed, which did not show statistical significance. This negative results could result from the limited sample size, the DNA sequencing process influenced by exterior factors and the geographical differences. Therefore, further analysis with a larger sample size or even a multicenter study is needed. And the association between the polymorphism of RS4360791 loci of ALOX5AP and ischemic stroke should be further analyzed to validate an early marker screening ischemic stroke, and to provide new insight of stroke prevention and treatment.

R e f e re n c e s [1] Izumimoto N, Kawakami A.Inflammation in atherosclerosis[J].Japanese Journal of Clinical Medicine,2011,69:2-5.
[2] Siasos G,Tsigkou V,Oikonomou E,et al.Circulating Biomarkers Determining Inflammation in Atherosclerosis Progression.[J].Current Medicinal Chemistry,2015,22(22):22-26.
[3] Spanbroek R, Gräbner R,Lötzer K, et al.Expanding expression of the 5-lipoxygenase pathway within the arterial wall during human atherogenesis[J].Proceedings of the National Academy of Sciences of the United States of America,2003,100(3):123-128.
[4] Nielsen M S,Gronholdt M L M,Vyberg M,et al.Adipose tissue arachidonic acid content is associated with the expression of 5-lipoxygenase in atherosclerotic plaques.[J].Lipids in Health