Displaying publications 1 - 20 of 109 in total

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  1. Vythilingam I, Chan ST, Shanmugratnam C, Tanrang H, Chooi KH
    Acta Trop, 2005 Oct;96(1):24-30.
    PMID: 16076459
    A study was carried out from July 2001 until January 2003 in the Kinabatangan area of Sabah, part of Borneo island, where malaria used to be mesoendemic. Vector surveys determined that Plasmodium falciparum was the predominant species and Anopheles balabacensis the primary vector. Malaria cases have dropped drastically over the years but P. falciparum is still predominant. In the present study, Anopheles donaldi was the predominant species and was positive for sporozoites. Although An. balabacensis was present, none were infective. An. donaldi bite more outdoors than indoors and have a peak biting time from 18:00 to 19:00 h when most people are still out of their homes. An integrated malaria control programme along with area development has helped in the control of malaria and its vector.
  2. Joanne S, Vythilingam I, Yugavathy N, Leong CS, Wong ML, AbuBakar S
    Acta Trop, 2015 Aug;148:38-45.
    PMID: 25899523 DOI: 10.1016/j.actatropica.2015.04.003
    Wolbachia are maternally transmitted bacteria found in most arthropods and nematodes, but little is known about their distribution and reproductive dynamics in the Malaysian dengue vector Aedes albopictus. In this study, polymerase chain reaction (PCR) was used to determine the presence of Wolbachia from field collected Ae. albopictus from various parts of the country using wsp specific primers. Ae. albopictus had Wolbachia infection ranging from 60 to 100%. No sequence diversity of wsp gene was found within all wAlbA and wAlbB sequences. Our findings suggest that Wolbachia infection amongst the Malaysian Ae. albopictus were not homogenously distributed in all districts in Malaysia. The presence of Wolbachia in different organs of Ae. albopictus was also determined. Wolbachia were only found in the ovaries and midguts of the mosquitoes, while absent in the salivary glands. The effects of Wolbachia on Ae. albopictus fecundity, longevity and egg viability were studied using infected and uninfected colonies. The removal of Wolbachia from Ae. albopictus resulted in reduced fecundity, longevity and egg viability, thus. Wolbachia seem to play a vital role in Ae. albopictus reproductive system.
  3. Low VL, Wong ML, Liew JWK, Pramasivan S, Jeyaprakasam NK, Vythilingam I
    Acta Trop, 2020 Jan;201:105207.
    PMID: 31586449 DOI: 10.1016/j.actatropica.2019.105207
    A gynandromorph of Culex sitiens Wiedemann (Diptera: Culicidae) was attracted to a human during a mosquito surveillance programme conducted in Kuala Lipis, Pahang, Malaysia on July 20, 2019. Gynandromorphism was observed in antennae, maxillary palps, legs and wings of the specimen, with distinct male characters on the left and female characters on the right, though the left maxillary palp is slightly shorter than the proboscis of a typical male. The abdomen, however, displays well-developed male genitalia. This study represents the first report of oblique gynandromorphism in Cx. sitiens, one of the vectors of Japanese encephalitis in Asia.
  4. Liew JWK, Selvarajoo S, Phang WK, Mah Hassan M, Redzuan MS, Selva Kumar S, et al.
    Acta Trop, 2021 Apr;216:105829.
    PMID: 33465350 DOI: 10.1016/j.actatropica.2021.105829
    The aim of this study is to investigate the feasibility and outcomes of using Gravid Oviposition Sticky (GOS) trap and dengue NS1 antigen tests for indoor and outdoor dengue/Aedes surveillance in the field. A one-year community-based study was carried out at Sungai Buloh Hospital Quarters, Selangor, Malaysia. GOS traps were first placed outdoors in three apartment blocks (Anggerik, Bunga Raya and Mawar). Beginning 29th week of the study, indoor traps were set in two apartment units on every floor in Anggerik. All female Aedes mosquitoes caught were tested for the presence of dengue NS1 antigen. Dengue seroprevalence and knowledge, attitude and practices on dengue prevention of the community and their reception to the surveillance approach were also assessed. Dengue-positive mosquitoes were detected at least 1 week before a dengue onset. More mosquitoes were caught indoors than outdoors in block Anggerik, but the total number of mosquitoes caught in all 3 blocks were similar. There was a significant difference in distribution of Ae. aegypti and Ae. albopictus between the 3 blocks. 66.1% and 3.4% of the community were positive for dengue IgG and IgM, respectively. Most respondents think that this surveillance method is Good (89%) and support its use nationwide. Dengue case ratio in the study apartment blocks decreased from year 2018 to 2019. This study demonstrated the practicality of performing proactive dengue/Aedes surveillance inside apartment units using the GOS traps. This surveillance method can be performed with immediate result output in the field.
  5. Leong CS, Vythilingam I, Wong ML, Wan Sulaiman WY, Lau YL
    Acta Trop, 2018 Sep;185:115-126.
    PMID: 29758171 DOI: 10.1016/j.actatropica.2018.05.008
    The resistance status of Selangor Aedes aegypti (Linnaeus) larvae against four major groups of insecticides (i.e., organochlorines, carbamates, organophosphates and pyrethroids) was investigated. Aedes aegypti were susceptible against temephos (organophosphate), although resistance (RR50 = 0.21-2.64) may be developing. The insecticides susceptibility status of Ae. aegypti larvae were found heterogeneous among the different study sites. Results showed that Ae. aegypti larvae from Klang, Sabak Bernam and Sepang were susceptible against all insecticides tested. However, other study sites exhibited low to high resistance against all pyrethroids (RR50 = 1.19-32.16). Overall, the application of synergists ethacrynic acid, S.S.S.- tributylphosphorotrithioate and piperonyl butoxide increased the toxicity of insecticides investigated. However, the application failed to increase the mortality to susceptible level (>97%) for certain populations, therefore there are chances of alteration of target site resistance involved. Biochemical assays revealed that α-esterase, (Gombak, Kuala Langat, Kuala Selangor and Sabak Bernam strains) β-esterase (Klang and Sabak Bernam strains), acetylcholinesterase (Kuala Selangor and Sabak Bernam strains), glutathione-S-transferase (Kuala Selangor and Sabak Bernam strains) and mono-oxygenases (Gombak, Hulu Langat, Hulu Selangor and Kuala Langat strains) were elevated. Spearman rank-order correlation indicated a significant correlation between resistance ratios of: DDT and deltamethrin (r = 0.683, P = 0.042), cyfluthrin and deltamethrin (r = 0.867, P =0.002), cyflyuthrin and lambdacyhalothrin (r = 0.800, P =0.010), cyfluthrin and permethrin (r = 0.770, P =0.015) deltamethrin and permethrin (r = 0.803, P =0.088), propoxur and malathion (r = 0.867, P = 0.002), malathion and temephos (r = 0.800, P = 0.010), etofenprox and MFO enzyme (r = 0.667, P =0.050). The current study provides baseline information for vector control programs conducted by local authorities. The susceptibility status of Ae. aegypti should be monitored sporadically to ensure the effectiveness of current vector control strategy in Selangor.
  6. Tan W, Liew JWK, Selvarajoo S, Lim XY, Foo CJ, Refai WF, et al.
    Acta Trop, 2020 Apr;204:105330.
    PMID: 31917959 DOI: 10.1016/j.actatropica.2020.105330
    The public health burden of dengue is most likely under reported. Current dengue control measures only considered symptomatic dengue transmission. Hence, there is a paucity of information on the epidemiology of inapparent dengue. This study reports that many people have been unknowingly exposed to dengue infection. Almost 10% and 70% of individuals without any history of dengue infection and living in a dengue hotspot, in Selangor, Malaysia, were dengue IgM and IgG positive respectively. When dengue-positive mosquitoes were detected in the hotspot, 11 (6.3%) of the 174 individuals tested were found to have dengue viremia, of which 10 were asymptomatic. Besides, upon detection of a dengue-infected mosquito, transmission was already widespread. In a clinical setting, it appears that people living with dengue patients have been exposed to dengue, whether asymptomatic or symptomatic. They can either have circulating viral RNA and/or presence of NS1 antigen. It is also possible that they are dengue seropositive. Collectively, the results indicate that actions taken to control dengue transmission after the first report of dengue cases may be already too late. The current study also revealed challenges in diagnosing clinically inapparent dengue in hyperendemic settings. There is no one best method for diagnosing inapparent dengue. This study demonstrates empirical evidence of inapparent dengue in different settings. Early dengue surveillance in the mosquito population and active serological/virological surveillance in humans can go hand in hand. More studies are required to investigate the epidemiology, seroprevalence, diagnostics, and control of inapparent dengue. It is also crucial to educate the public, health staff and medical professionals on asymptomatic dengue and to propagate awareness, which is important for controlling transmission.
  7. Ong NH, Chua CL, Liew JWK, Wan Sulaiman WY, Chan YF, Sam IC, et al.
    Acta Trop, 2020 Aug;208:105472.
    PMID: 32389451 DOI: 10.1016/j.actatropica.2020.105472
    Zika virus (ZIKV) is a mosquito-borne flavivirus with global impact since 2015. Although ZIKV was first isolated from Aedes aegypti in Malaysia in 1965, not much is known about the competency of Malaysian Ae. aegypti to ZIKV. To date only 9 cases of ZIKV have been reported in Malaysia despite the abundance of mosquito vectors. This study aimed to determine the susceptibility of Ae. aegypti to ZIKV, and the impact of sequential infections in Ae. aegypti mosquitoes with DENV serotype 2 (DENV-2) followed by ZIKV. Field-caught urban Ae. aegypti were orally challenged with a Martinique strain of ZIKV, and midgut, head/thorax and saliva were collected at 3, 7 and 14 days post-infection (dpi). At 14 dpi, ZIKV-exposed mosquitoes had infection and dissemination rates of 59% (n=10/17) and 90% (n=9/10), respectively. Average titres of 3.9 and 4.4 log pfu infectious ZIKV were recovered in midgut and head/thorax, respectively. In sequential infection, prior exposure of Ae. aegypti to DENV did not affect the subsequent ZIKV infection in head/thorax albeit with a low sample size. In conclusion, Malaysian urban Ae. aegypti is susceptible to the contemporary Asian lineage of ZIKV. The established and continuous DENV circulation in Ae. aegypti did not suppress ZIKV emergence in Malaysia. Other factors contributing to low level of ZIKV circulation in Malaysia remain to be explored.
  8. Vythilingam I, Chua TH, Liew JWK, Manin BO, Ferguson HM
    Adv Parasitol, 2021;113:131-189.
    PMID: 34620382 DOI: 10.1016/bs.apar.2021.08.005
    Plasmodium knowlesi, a simian malaria parasite of great public health concern has been reported from most countries in Southeast Asia and exported to various countries around the world. Currently P. knowlesi is the predominant species infecting humans in Malaysia. Besides this species, other simian malaria parasites such as P. cynomolgi and P. inui are also infecting humans in the region. The vectors of P. knowlesi and other Asian simian malarias belong to the Leucosphyrus Group of Anopheles mosquitoes which are generally forest dwelling species. Continual deforestation has resulted in these species moving into forest fringes, farms, plantations and human settlements along with their macaque hosts. Limited studies have shown that mosquito vectors are attracted to both humans and macaque hosts, preferring to bite outdoors and in the early part of the night. We here review the current status of simian malaria vectors and their parasites, knowledge of vector competence from experimental infections and discuss possible vector control measures. The challenges encountered in simian malaria elimination are also discussed. We highlight key knowledge gaps on vector distribution and ecology that may impede effective control strategies.
  9. Joanne S, Vythilingam I, Yugavathy N, Doss JI
    Asian Pac J Trop Biomed, 2014 Jul;4(7):557-60.
    PMID: 25183276 DOI: 10.12980/APJTB.4.2014APJTB-2014-0020
    To develop an artificial and modified Wolbachia removal technique using tetracycline from naturally Wolbachia infected Aedes albopictus (Ae. albopictus) so as to be able to produce generations of Wolbachia free offsprings.
  10. Yap NJ, Hossain H, Nada-Raja T, Ngui R, Muslim A, Hoh BP, et al.
    Emerg Infect Dis, 2021 08;27(8):2187-2191.
    PMID: 34287122 DOI: 10.3201/eid2708.204502
    We detected the simian malaria parasites Plasmodium knowlesi, P. cynomolgi, P. inui, P. coatneyi, P. inui-like, and P. simiovale among forest fringe-living indigenous communities from various locations in Malaysia. Our findings underscore the importance of using molecular tools to identify newly emergent malaria parasites in humans.
  11. Liew JWK, Bukhari FDM, Jeyaprakasam NK, Phang WK, Vythilingam I, Lau YL
    Emerg Infect Dis, 2021 10;27(10):2700-2703.
    PMID: 34545786 DOI: 10.3201/eid2710.210412
    We detected 2 natural, asymptomatic Plasmodium inui monoinfections in humans in Malaysia by using nested PCR on concentrated high-volume blood samples. We found a P. inui-positive Anopheles cracens mosquito in the same site as the human infections. Investigators should use ultrasensitive detection methods to identify simian malaria parasite transmission in humans.
  12. Anderios F, Noorrain A, Vythilingam I
    Exp Parasitol, 2010 Feb;124(2):181-9.
    PMID: 19765587 DOI: 10.1016/j.exppara.2009.09.009
    Plasmodium knowlesi is a malaria parasite of Old World monkeys and is infectious to humans. In this study Macaca fascicularis was used as a model to understand the host response to P. knowlesi using parasitological and haematological parameters. Three M. fascicularis of either sex were experimentally infected with P. knowlesi erythrocytic parasites from humans. The pre-patent period for P. knowlesi infection in M. fascicularis ranged from seven to 14 days. The parasitemia observed was 13,686-24,202 parasites per microL of blood for asexual stage and 88-264 parasites per microL of blood for sexual stage. Periodicity analysis adopted from microfilaria periodicity technique of asexual stage showed that the parasitemia peak at 17:39h while the sexual stage peaked at 02:36 h. Mathematical analysis of the data indicates that P. knowlesi gametocytes tend to display periodicity with a peak (24:00-06:00) that coincides with the peak biting activity (19:00-06:00) of the local vector, Anopheles latens. The morphology of P. knowlesi resembled P. falciparum in early trophozoite and P. malariae in late trophozoite. However, it may be distinguishable by observing the appliqué appearance of the cytoplasm and the chromatin lying inside the ring. Haematological analysis on macaques with knowlesi malaria showed clinical manifestations of hypoglycaemia, anaemia and hyperbilirubinemia. Gross examination of spleen and liver showed malaria pigments deposition in both organs.
  13. Vythilingam I, Sam JI, Chan YF, Khaw LT, Sulaiman WY
    Front Microbiol, 2016;7:1452.
    PMID: 27679623 DOI: 10.3389/fmicb.2016.01452
    Zika virus (ZIKV) has now become a global public health concern. The vectors for ZIKV are Aedes aegypti and A. albopictus. Both these mosquitoes are predominant in Southeast Asia and are also responsible for the spread of other arboviral diseases like dengue virus and chikungunya virus. The incidence of dengue has been increasing over the years and this is of concern to public health workers. Simple laboratory tools for the detection of ZIKV is also lacking. In the absence of drugs and vaccine for these arboviral diseases, vector control is the main option for surveillance and control. Aedes larval surveys have been the hallmark of dengue control along with larviciding and fogging when cases are reported. However, we need new paradigms and options for control of these vectors. The current situation in Southeast Asia clearly proves that effective strategies for vector control need to be proactive and not reactive. This will be the way forward to control epidemics of these diseases inclusive of ZIKV until a vaccine becomes available.
  14. Wong ML, Zulzahrin Z, Vythilingam I, Lau YL, Sam IC, Fong MY, et al.
    Front Microbiol, 2023;14:1135977.
    PMID: 37025644 DOI: 10.3389/fmicb.2023.1135977
    The complex transmission profiles of vector-borne zoonoses (VZB) and vector-borne infections with animal reservoirs (VBIAR) complicate efforts to break the transmission circuit of these infections. To control and eliminate VZB and VBIAR, insecticide application may not be conducted easily in all circumstances, particularly for infections with sylvatic transmission cycle. As a result, alternative approaches have been considered in the vector management against these infections. In this review, we highlighted differences among the environmental, chemical, and biological control approaches in vector management, from the perspectives of VZB and VBIAR. Concerns and knowledge gaps pertaining to the available control approaches were discussed to better understand the prospects of integrating these vector control approaches to synergistically break the transmission of VZB and VBIAR in humans, in line with the integrated vector management (IVM) developed by the World Health Organization (WHO) since 2004.
  15. Vythilingam I
    Front Physiol, 2012;3:115.
    PMID: 22557977 DOI: 10.3389/fphys.2012.00115
    Malaria and filariasis still continue to pose public health problems in developing countries of the tropics. Although plans are in progress for the elimination of both these parasitic vector borne diseases, we are now faced with a daunting challenge as we have a fifth species, Plasmodium knowlesi a simian malaria parasite affecting humans. Similarly in peninsular Malaysia, filariasis was mainly due to Brugia malayi. However, we now see cases of Wuchereria bancrofti in immigrant workers coming into the country. In order to successfully eliminate both these diseases we need to know the vectors involved and introduce appropriate control measures to prevent the diseases occurring in the future. As for knowlesi malaria it is still uncertain if human to human transmission through mosquito bites is occurring. However, P. knowlesi in human is not a rare occurrence anymore and has all the characteristics of a pathogen spreading due to changes in the ecosystem, international travel, and cross border migration. This has created a more complex situation. In order to overcome these challenges we need to revamp our control measures. This paper reviews the vectors of malaria and filariasis in Southeast Asia with special emphasis on P. knowlesi and W. bancrofti in Malaysia and their control strategies.
  16. Zulzahrin Z, Wong ML, Naziri MRA, Lau YL, Vythilingam I, Lee WC
    Heliyon, 2024 Feb 15;10(3):e25207.
    PMID: 38322922 DOI: 10.1016/j.heliyon.2024.e25207
    Wing measurement is an important parameter in many entomological studies. However, the methods of measuring wings vary with studies, and a gold standard method was not available for this procedure. This in turn limits researchers from confidently comparing their research findings with published data collected by other means of measurement. This study investigated the interchangeability of three commonly available methods for wing measurement, namely the calliper method, stereomicroscope-assisted photography method, and digital microscope-assisted photography method, using the laboratory colony of Aedes aegypti. It was found that the calliper method and the photography-based methods yielded similar results, hence the good interchangeability of these methods. Nevertheless, the digital microscope-assisted photography method yielded more accurate measurements, due to the higher resolution of the captured photos, and minimal technical bias during the data collection, as compared to the calliper-based and stereomicroscope-assisted photography methods. This study served as a reference for researchers to select the most suitable measurement method in future studies.
  17. Wong ML, Ahmed MA, Sulaiman WYW, Manin BO, Leong CS, Quan FS, et al.
    Infect Genet Evol, 2019 09;73:26-32.
    PMID: 30999059 DOI: 10.1016/j.meegid.2019.04.010
    We explored and constructed haplotype network for simian malaria species: Plasmodium knowlesi, P. cynomolgi and P. inui aiming to understand the transmission dynamics between mosquitoes, humans and macaques. Mosquitoes were collected from villages in an area where zoonotic malaria is prevalent. PCR analysis confirmed Anopheles balabacensis as the main vector for macaque parasites, moreover nearly 60% of the mosquitoes harboured more than one Plasmodium species. Fragments of the A-type small subunit ribosomal RNA (SS rRNA) amplified from salivary gland sporozoites, and equivalent sequences obtained from GenBank were used to construct haplotype networks. The patterns were consistent with the presence of geographically distinct populations for P. inui and P. cynomolgi, and with three discrete P. knowlesi populations. This study provides a preliminary snapshot of the structure of these populations, that was insufficient to answer our aim. Thus, collection of parasites from their various hosts and over time, associated with a systematic analysis of a set of genetical loci is strongly advocated in order to obtain a clear picture of the parasite population and the flow between different hosts. This is important to devise measures that will minimise the risk of transmission to humans, because zoonotic malaria impedes malaria elimination.
  18. Yap NJ, Goh XT, Koehler AV, William T, Yeo TW, Vythilingam I, et al.
    Infect Genet Evol, 2017 10;54:39-46.
    PMID: 28634105 DOI: 10.1016/j.meegid.2017.06.019
    Plasmodium knowlesi, a malaria parasite of macaques, has emerged as an important parasite of humans. Despite the significance of P. knowlesi malaria in parts of Southeast Asia, very little is known about the genetic variation in this parasite. Our aim here was to explore sequence variation in a molecule called the 42kDa merozoite surface protein-1 (MSP-1), which is found on the surface of blood stages of Plasmodium spp. and plays a key role in erythrocyte invasion. Several studies of P. falciparum have reported that the C-terminus (a 42kDa fragment) of merozoite surface protein-1 (MSP-142; consisting of MSP-119 and MSP-133) is a potential candidate for a malaria vaccine. However, to date, no study has yet investigated the sequence diversity of the gene encoding P. knowlesi MSP-142 (comprising Pk-msp-119 and Pk-msp-133) among isolates in Malaysia. The present study explored this aspect. Twelve P. knowlesi isolates were collected from patients from hospitals in Selangor and Sabah Borneo, Malaysia, between 2012 and 2014. The Pk-msp-142 gene was amplified by PCR and directly sequenced. Haplotype diversity (Hd) and nucleotide diversity (л) were studied among the isolates. There was relatively high genetic variation among P. knowlesi isolates; overall Hd and л were 1±0.034 and 0.01132±0.00124, respectively. A total of nine different haplotypes related to amino acid alterations at 13 positions, and the Pk-MSP-119 sequence was found to be more conserved than Pk-msp-133. We have found evidence for negative selection in Pk-msp-42 as well as the 33kDa and 19kDa fragments by comparing the rate of non-synonymous versus synonymous substitutions. Future investigations should study large numbers of samples from disparate geographical locations to critically assess whether this molecule might be a potential vaccine target for P. knowlesi.
  19. Liew JWK, Selvarajoo S, Tan W, Ahmad Zaki R, Vythilingam I
    Infect Dis Poverty, 2019 Sep 03;8(1):71.
    PMID: 31477185 DOI: 10.1186/s40249-019-0584-y
    BACKGROUND: Dengue is a global disease, transmitted by the Aedes vectors. In 2018, there were 80 615 dengue cases with 147 deaths in Malaysia. Currently, the nationwide surveillance programs are dependent on Aedes larval surveys and notifications of lab-confirmed human infections. The existing, reactive programs appear to lack sensitivity and proactivity. More efficient dengue vector surveillance/control methods are needed.

    METHODS: A parallel, cluster, randomized controlled, interventional trial is being conducted for 18 months in Damansara Damai, Selangor, Malaysia, to determine the efficacy of using gravid oviposition sticky (GOS) trap and dengue non-structural 1 (NS1) antigen test for early surveillance of dengue among Aedes mosquitoes to reduce dengue outbreaks. Eight residential apartments were randomly assigned into intervention and control arms. GOS traps are set at the apartments to collect Aedes weekly, following which dengue NS1 antigen is detected in these mosquitoes. When a dengue-positive mosquito is detected, the community will be advised to execute vector search-and-destroy and protective measures. The primary outcome concerns the the percentage change in the (i) number of dengue cases and (ii) durations of dengue outbreaks. Whereas other outcome measures include the change in density threshold of Aedes and changes in dengue-related knowledge, attitude and practice among cluster inhabitants.

    DISCUSSION: This is a proactive and early dengue surveillance in the mosquito vector that does not rely on notification of dengue cases. Surveillance using the GOS traps should be able to efficiently provide sufficient coverage for multistorey dwellings where population per unit area is likely to be higher. Furthermore, trapping dengue-infected mosquitoes using the GOS trap, helps to halt the dengue transmission carried by the mosquito. It is envisaged that the results of this randomized controlled trial will provide a new proactive, cheap and targeted surveillance tool for the prevention and control of dengue outbreaks.

    TRIAL REGISTRATION: This is a parallel-cluster, randomized controlled, interventional trial, registered at ClinicalTrials.gov (ID: NCT03799237), on 8th January 2019 (retrospectively registered).

  20. Boyle JH, Rastas PMA, Huang X, Garner AG, Vythilingam I, Armbruster PA
    Insects, 2021 Feb 16;12(2).
    PMID: 33669192 DOI: 10.3390/insects12020167
    The Asian tiger mosquito, Aedes albopictus, is an invasive vector mosquito of substantial public health concern. The large genome size (~1.19-1.28 Gb by cytofluorometric estimates), comprised of ~68% repetitive DNA sequences, has made it difficult to produce a high-quality genome assembly for this species. We constructed a high-density linkage map for Ae. albopictus based on 111,328 informative SNPs obtained by RNAseq. We then performed a linkage-map anchored reassembly of AalbF2, the genome assembly produced by Palatini et al. (2020). Our reassembled genome sequence, AalbF3, represents several improvements relative to AalbF2. First, the size of the AalbF3 assembly is 1.45 Gb, almost half the size of AalbF2. Furthermore, relative to AalbF2, AalbF3 contains a higher proportion of complete and single-copy BUSCO genes (84.3%) and a higher proportion of aligned RNAseq reads that map concordantly to a single location of the genome (46%). We demonstrate the utility of AalbF3 by using it as a reference for a bulk-segregant-based comparative genomics analysis that identifies chromosomal regions with clusters of candidate SNPs putatively associated with photoperiodic diapause, a crucial ecological adaptation underpinning the rapid range expansion and climatic adaptation of A. albopictus.
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