Displaying publications 1 - 20 of 110 in total

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  1. Sukumarran D, Hasikin K, Khairuddin ASM, Ngui R, Sulaiman WYW, Vythilingam I, et al.
    Parasit Vectors, 2024 Apr 16;17(1):188.
    PMID: 38627870 DOI: 10.1186/s13071-024-06215-7
    BACKGROUND: Malaria is a serious public health concern worldwide. Early and accurate diagnosis is essential for controlling the disease's spread and avoiding severe health complications. Manual examination of blood smear samples by skilled technicians is a time-consuming aspect of the conventional malaria diagnosis toolbox. Malaria persists in many parts of the world, emphasising the urgent need for sophisticated and automated diagnostic instruments to expedite the identification of infected cells, thereby facilitating timely treatment and reducing the risk of disease transmission. This study aims to introduce a more lightweight and quicker model-but with improved accuracy-for diagnosing malaria using a YOLOv4 (You Only Look Once v. 4) deep learning object detector.

    METHODS: The YOLOv4 model is modified using direct layer pruning and backbone replacement. The primary objective of layer pruning is the removal and individual analysis of residual blocks within the C3, C4 and C5 (C3-C5) Res-block bodies of the backbone architecture's C3-C5 Res-block bodies. The CSP-DarkNet53 backbone is simultaneously replaced for enhanced feature extraction with a shallower ResNet50 network. The performance metrics of the models are compared and analysed.

    RESULTS: The modified models outperform the original YOLOv4 model. The YOLOv4-RC3_4 model with residual blocks pruned from the C3 and C4 Res-block body achieves the highest mean accuracy precision (mAP) of 90.70%. This mAP is > 9% higher than that of the original model, saving approximately 22% of the billion floating point operations (B-FLOPS) and 23 MB in size. The findings indicate that the YOLOv4-RC3_4 model also performs better, with an increase of 9.27% in detecting the infected cells upon pruning the redundant layers from the C3 Res-block bodies of the CSP-DarkeNet53 backbone.

    CONCLUSIONS: The results of this study highlight the use of the YOLOv4 model for detecting infected red blood cells. Pruning the residual blocks from the Res-block bodies helps to determine which Res-block bodies contribute the most and least, respectively, to the model's performance. Our method has the potential to revolutionise malaria diagnosis and pave the way for novel deep learning-based bioinformatics solutions. Developing an effective and automated process for diagnosing malaria will considerably contribute to global efforts to combat this debilitating disease. We have shown that removing undesirable residual blocks can reduce the size of the model and its computational complexity without compromising its precision.

  2. 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.
  3. Pramasivan S, Ngui R, Jeyaprakasam NK, Low VL, Liew JWK, Vythilingam I
    Parasit Vectors, 2023 Oct 09;16(1):355.
    PMID: 37814287 DOI: 10.1186/s13071-023-05984-x
    BACKGROUND: Malaria parasites such as Plasmodium knowlesi, P. inui, and P. cynomolgi are spread from macaques to humans through the Leucosphyrus Group of Anopheles mosquitoes. It is crucial to know the distribution of these vectors to implement effective control measures for malaria elimination. Plasmodium knowlesi is the most predominant zoonotic malaria parasite infecting humans in Malaysia.

    METHODS: Vector data from various sources were used to create distribution maps from 1957 to 2021. A predictive statistical model utilizing logistic regression was developed using significant environmental factors. Interpolation maps were created using the inverse distance weighted (IDW) method and overlaid with the corresponding environmental variables.

    RESULTS: Based on the IDW analysis, high vector abundances were found in the southwestern part of Sarawak, the northern region of Pahang and the northwestern part of Sabah. However, most parts of Johor, Sabah, Perlis, Penang, Kelantan and Terengganu had low vector abundance. The accuracy test indicated that the model predicted sampling and non-sampling areas with 75.3% overall accuracy. The selected environmental variables were entered into the regression model based on their significant values. In addition to the presence of water bodies, elevation, temperature, forest loss and forest cover were included in the final model since these were significantly correlated. Anopheles mosquitoes were mainly distributed in Peninsular Malaysia (Titiwangsa range, central and northern parts), Sabah (Kudat, West Coast, Interior and Tawau division) and Sarawak (Kapit, Miri, and Limbang). The predicted Anopheles mosquito density was lower in the southern part of Peninsular Malaysia, the Sandakan Division of Sabah and the western region of Sarawak.

    CONCLUSION: The study offers insight into the distribution of the Leucosphyrus Group of Anopheles mosquitoes in Malaysia. Additionally, the accompanying predictive vector map correlates well with cases of P. knowlesi malaria. This research is crucial in informing and supporting future efforts by healthcare professionals to develop effective malaria control interventions.

  4. Fu JYL, Chua CL, Abu Bakar AS, Vythilingam I, Wan Sulaiman WY, Alphey L, et al.
    PLoS Negl Trop Dis, 2023 Jun;17(6):e0011423.
    PMID: 37307291 DOI: 10.1371/journal.pntd.0011423
    BACKGROUND: Emerging arboviruses such as chikungunya and Zika viruses have unexpectedly caused widespread outbreaks in tropical and subtropical regions recently. Ross River virus (RRV) is endemic in Australia and has epidemic potential. In Malaysia, Aedes mosquitoes are abundant and drive dengue and chikungunya outbreaks. We assessed risk of an RRV outbreak in Kuala Lumpur, Malaysia by determining vector competence of local Aedes mosquitoes and local seroprevalence as a proxy of human population susceptibility.

    METHODOLOGY/PRINCIPAL FINDINGS: We assessed oral susceptibility of Malaysian Ae. aegypti and Ae. albopictus by real-time PCR to an Australian RRV strain SW2089. Replication kinetics in midgut, head and saliva were determined at 3 and 10 days post-infection (dpi). With a 3 log10 PFU/ml blood meal, infection rate was higher in Ae. albopictus (60%) than Ae. aegypti (15%; p<0.05). Despite similar infection rates at 5 and 7 log10 PFU/ml blood meals, Ae. albopictus had significantly higher viral loads and required a significantly lower median oral infectious dose (2.7 log10 PFU/ml) than Ae. aegypti (4.2 log10 PFU/ml). Ae. albopictus showed higher vector competence, with higher viral loads in heads and saliva, and higher transmission rate (RRV present in saliva) of 100% at 10 dpi, than Ae. aegypti (41%). Ae. aegypti demonstrated greater barriers at either midgut escape or salivary gland infection, and salivary gland escape. We then assessed seropositivity against RRV among 240 Kuala Lumpur inpatients using plaque reduction neutralization, and found a low rate of 0.8%.

    CONCLUSIONS/SIGNIFICANCE: Both Ae. aegypti and Ae. albopictus are susceptible to RRV, but Ae. albopictus displays greater vector competence. Extensive travel links with Australia, abundant Aedes vectors, and low population immunity places Kuala Lumpur, Malaysia at risk of an imported RRV outbreak. Surveillance and increased diagnostic awareness and capacity are imperative to prevent establishment of new arboviruses in Malaysia.

  5. Jeyaprakasam NK, Low VL, Pramasivan S, Liew JWK, Wan-Sulaiman WY, Vythilingam I
    PLoS Negl Trop Dis, 2023 Jun;17(6):e0011438.
    PMID: 37384790 DOI: 10.1371/journal.pntd.0011438
    BACKGROUND: The elimination of malaria in Southeast Asia has become more challenging as a result of rising knowlesi malaria cases. In addition, naturally occurring human infections with other zoonotic simian malaria caused by Plasmodium cynomolgi and Plasmodium inui adds another level of complexity in malaria elimination in this region. Unfortunately, data on vectors which are responsible for transmitting this zoonotic disease is very limited.

    METHODOLOGY/PRINCIPAL FINDINGS: We conducted longitudinal studies to investigate the entomological parameters of the simian malaria vectors and to examine the genetic diversity and evolutionary pattern of their simian Plasmodium. All the captured Anopheles mosquitoes were dissected to examine for the presence of oocysts, sporozoites and to determine the parous rate. Our study revealed that the Anopheles Leucosphyrus Group mosquitoes are highly potential competent vectors, as evidenced by their high rate of parity, survival and sporozoite infections in these mosquitoes. Thus, these mosquitoes represent a risk of human infection with zoonotic simian malaria in this region. Haplotype analysis on P. cynomolgi and P. inui, found in high prevalence in the Anopheles mosquitoes from this study, had shown close relationship between simian Plasmodium from the Anopheles mosquitoes with its vertebrate hosts. This directly signifies the ongoing transmission between the vector, macaques, and humans. Furthermore, population genetic analysis showed significant negative values which suggest that both Plasmodium species are undergoing population expansion.

    CONCLUSIONS/SIGNIFICANCE: With constant microevolutionary processes, there are potential for both P. inui and P. cynomolgi to emerge and spread as a major public health problem, following the similar trend of P. knowlesi. Therefore, concerted vector studies in other parts of Southeast Asia are warranted to better comprehend the transmission dynamics of this zoonotic simian malaria which eventually would aid in the implementation of effective control measures in a rapidly changing environment.

  6. Sukumarran D, Hasikin K, Mohd Khairuddin AS, Ngui R, Wan Sulaiman WY, Vythilingam I, et al.
    Trop Biomed, 2023 Jun 01;40(2):208-219.
    PMID: 37650409 DOI: 10.47665/tb.40.2.013
    Timely and rapid diagnosis is crucial for faster and proper malaria treatment planning. Microscopic examination is the gold standard for malaria diagnosis, where hundreds of millions of blood films are examined annually. However, this method's effectiveness depends on the trained microscopist's skills. With the increasing interest in applying deep learning in malaria diagnosis, this study aims to determine the most suitable deep-learning object detection architecture and their applicability to detect and distinguish red blood cells as either malaria-infected or non-infected cells. The object detectors Yolov4, Faster R-CNN, and SSD 300 are trained with images infected by all five malaria parasites and from four stages of infection with 80/20 train and test data partition. The performance of object detectors is evaluated, and hyperparameters are optimized to select the best-performing model. The best-performing model was also assessed with an independent dataset to verify the models' ability to generalize in different domains. The results show that upon training, the Yolov4 model achieves a precision of 83%, recall of 95%, F1-score of 89%, and mean average precision of 93.87% at a threshold of 0.5. Conclusively, Yolov4 can act as an alternative in detecting the infected cells from whole thin blood smear images. Object detectors can complement a deep learning classification model in detecting infected cells since they eliminate the need to train on single-cell images and have been demonstrated to be more feasible for a different target domain.
  7. 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.
  8. Roslan MA, Ngui R, Vythilingam I, Wan Sulaiman WY
    J Vector Ecol, 2022 Dec;47(2):142-152.
    PMID: 36314668 DOI: 10.52707/1081-1710-47.2.142
    The study assessed the distribution of Malaysian Ae. albopictus adults associated with Wolbachia detection in low-rise residential areas using a modified sticky ovitrap (MSO). The relationship between Ae. albopictus and climatological parameters were also determined. Fifty-two weeks of surveillance using 273 MSOs were conducted in four installation areas of eleven sampling sites. Specimens were subjected to PCR using wsp-specific primers for Wolbachia detection. The relationship between climatological parameters and Ae. albopictus captured were analyzed using Spearman rank correlation coefficient test. The majority of Ae. albopictus were captured in residential houses (87%), followed by playgrounds or parks (11.5%), guardhouses (1%), and community halls (0.5%). Most of the specimens (92%) were superinfected with wAlbA and wAlbB strains. A positive correlation with no significant association was found for rainfall (r = 0.015, P = 0.072), relative humidity (r = 0.005, P = 0.526), minimum temperature (r = 0.005, P = 0.516), and mean temperature (r = 0.003, P = 0.689). MSO effectively captured a high number of Ae. albopictus that was determined to be the predominant mosquito species found in low-rise residential areas. The adult collection is not only influenced by climatological parameters but also by other factors, including environmental conditions and general sanitation status.
  9. Selvarajoo S, Liew JWK, Chua TH, Tan W, Zaki RA, Ngui R, et al.
    Sci Rep, 2022 01 12;12(1):571.
    PMID: 35022501 DOI: 10.1038/s41598-021-04643-4
    Dengue remains a major public threat and existing dengue control/surveillance programs lack sensitivity and proactivity. More efficient methods are needed. A cluster randomized controlled trial was conducted for 18 months to determine the efficacy of using a combination of gravid oviposition sticky (GOS) traps and dengue non-structural 1 (NS1) antigen for early surveillance of dengue among Aedes mosquito. Eight residential apartments were randomly assigned into intervention and control groups. GOS traps were placed at the intervention apartments weekly to trap Aedes mosquitoes and these tested for dengue NS1 antigen. When dengue-positive pool was detected, the community were notified and advised to execute protective measures. Fewer dengue cases were recorded in the intervention group than the control. Detection of NS1-positive mosquitoes was significantly associated with GOS Aedes index (rs = 0.68, P 
  10. Jeyaprakasam NK, Low VL, Liew JWK, Pramasivan S, Wan-Sulaiman WY, Saeung A, et al.
    Sci Rep, 2022 01 10;12(1):354.
    PMID: 35013403 DOI: 10.1038/s41598-021-04106-w
    Blood feeding and host-seeking behaviors of a mosquito play an imperative role in determining its vectorial capacity in transmitting pathogens. Unfortunately, limited information is available regarding blood feeding behavior of Anopheles species in Malaysia. Collection of resting Anopheles mosquitoes for blood meal analysis poses a great challenge especially for forest dwelling mosquitoes. Therefore, a laboratory-based study was conducted to evaluate the potential use of mosquitoes caught using human landing catch (HLC) for blood meal analysis, and subsequently to document blood feeding behavior of local Anopheles mosquitoes in Peninsular Malaysia. The laboratory-based experiment from this study revealed that mosquitoes caught using HLC had the potential to be used for blood meal analysis. Besides HLC, mosquitoes were also collected using manual aspirator and Mosquito Magnet. Overall, 47.4% of 321 field-caught Anopheles mosquitoes belonging to six species were positive for vertebrate host DNA in their blood meal. The most frequent blood meal source was human (45.9%) followed by wild boar (27.4%), dog (15.3%) and monkey (7.5%). Interestingly, only Anopheles cracens and Anopheles introlatus (Leucosphyrus Group) fed on monkey. This study further confirmed that members of the Leucosphyrus Group are the predominant vectors for knowlesi malaria transmission in Peninsular Malaysia mainly due to their simio-anthropophagic feeding behavior.
  11. Pramasivan S, Ngui R, Jeyaprakasam NK, Liew JWK, Low VL, Mohamed Hassan N, et al.
    Malar J, 2021 Oct 29;20(1):426.
    PMID: 34715864 DOI: 10.1186/s12936-021-03963-0
    BACKGROUND: Plasmodium knowlesi, a simian malaria parasite infection, increases as Plasmodium falciparum and Plasmodium vivax infections decrease in Johor, Malaysia. Therefore, this study aimed to identify the distribution of vectors involved in knowlesi malaria transmission in Johor. This finding is vital in estimating hotspot areas for targeted control strategies.

    METHODS: Anopheles mosquitoes were collected from the location where P. knowlesi cases were reported. Cases of knowlesi malaria from 2011 to 2019 in Johor were analyzed. Internal transcribed spacers 2 (ITS2) and cytochrome c oxidase subunit I (COI) genes were used to identify the Leucosphyrus Group of Anopheles mosquitoes. In addition, spatial analysis was carried out on the knowlesi cases and vectors in Johor.

    RESULTS: One hundred and eighty-nine cases of P. knowlesi were reported in Johor over 10 years. Young adults between the ages of 20-39 years comprised 65% of the cases. Most infected individuals were involved in agriculture and army-related occupations (22% and 32%, respectively). Four hundred and eighteen Leucosphyrus Group Anopheles mosquitoes were captured during the study. Anopheles introlatus was the predominant species, followed by Anopheles latens. Spatial analysis by Kriging interpolation found that hotspot regions of P. knowlesi overlapped or were close to the areas where An. introlatus and An. latens were found. A significantly high number of vectors and P. knowlesi cases were found near the road within 0-5 km.

    CONCLUSIONS: This study describes the distribution of P. knowlesi cases and Anopheles species in malaria-endemic transmission areas in Johor. Geospatial analysis is a valuable tool for studying the relationship between vectors and P. knowlesi cases. This study further supports that the Leucosphyrus Group of mosquitoes might be involved in transmitting knowlesi malaria cases in Johor. These findings may provide initial evidence to prioritize diseases and vector surveillance.

  12. 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.
  13. 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.
  14. Byrne I, Aure W, Manin BO, Vythilingam I, Ferguson HM, Drakeley CJ, et al.
    Sci Rep, 2021 Jun 03;11(1):11810.
    PMID: 34083582 DOI: 10.1038/s41598-021-90893-1
    Land-use changes, such as deforestation and agriculture, can influence mosquito vector populations and malaria transmission. These land-use changes have been linked to increased incidence in human cases of the zoonotic malaria Plasmodium knowlesi in Sabah, Malaysian Borneo. This study investigates whether these associations are partially driven by fine-scale land-use changes creating more favourable aquatic breeding habitats for P. knowlesi anopheline vectors. Using aerial remote sensing data, we developed a sampling frame representative of all land use types within a major focus of P. knowlesi transmission. From 2015 to 2016 monthly longitudinal surveys of larval habitats were collected in randomly selected areas stratified by land use type. Additional remote sensing data on environmental variables, land cover and landscape configuration were assembled for the study site. Risk factor analyses were performed over multiple spatial scales to determine associations between environmental and spatial variables and anopheline larval presence. Habitat fragmentation (300 m), aspect (350 m), distance to rubber plantations (100 m) and Culex larval presence were identified as risk factors for Anopheles breeding. Additionally, models were fit to determine the presence of potential larval habitats within the areas surveyed and used to generate a time-series of monthly predictive maps. These results indicate that land-use change and topography influence the suitability of larval habitats, and may partially explain the link between P. knowlesi incidence and deforestation. The predictive maps, and identification of the spatial scales at which risk factors are most influential may aid spatio-temporally targeted vector control interventions.
  15. 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.
  16. Jeyaprakasam NK, Pramasivan S, Liew JWK, Van Low L, Wan-Sulaiman WY, Ngui R, et al.
    Parasit Vectors, 2021 Apr 01;14(1):184.
    PMID: 33794965 DOI: 10.1186/s13071-021-04689-3
    BACKGROUND: Vector surveillance is essential in determining the geographical distribution of mosquito vectors and understanding the dynamics of malaria transmission. With the elimination of human malaria cases, knowlesi malaria cases in humans are increasing in Malaysia. This necessitates intensive vector studies using safer trapping methods which are both field efficient and able to attract the local vector populations. Thus, this study evaluated the potential of Mosquito Magnet as a collection tool for Anopheles mosquito vectors of simian malaria along with other known collection methods.

    METHODS: A randomized 4 × 4 Latin square designed experiment was conducted to compare the efficiency of the Mosquito Magnet against three other common trapping methods: human landing catch (HLC), CDC light trap and human baited trap (HBT). The experiment was conducted over six replicates where sampling within each replicate was carried out for 4 consecutive nights. An additional 4 nights of sampling was used to further evaluate the Mosquito Magnet against the "gold standard" HLC. The abundance of Anopheles sampled by different methods was compared and evaluated with focus on the Anopheles from the Leucosphyrus group, the vectors of knowlesi malaria.

    RESULTS: The Latin square designed experiment showed HLC caught the greatest number of Anopheles mosquitoes (n = 321) compared to the HBT (n = 87), Mosquito Magnet (n = 58) and CDC light trap (n = 13). The GLMM analysis showed that the HLC method caught significantly more Anopheles mosquitoes compared to Mosquito Magnet (P = 0.049). However, there was no significant difference in mean nightly catch of Anopheles mosquitoes between Mosquito Magnet and the other two trapping methods, HBT (P = 0.646) and CDC light traps (P = 0.197). The mean nightly catch for both An. introlatus (9.33 ± 4.341) and An. cracens (4.00 ± 2.273) caught using HLC was higher than that of Mosquito Magnet, though the differences were not statistically significant (P > 0.05). This is in contrast to the mean nightly catch of An. sinensis (15.75 ± 5.640) and An. maculatus (15.78 ± 3.479) where HLC showed significantly more mosquito catches compared to Mosquito Magnet (P 

  17. 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.
  18. Li MI, Mailepessov D, Vythilingam I, Lee V, Lam P, Ng LC, et al.
    PLoS Negl Trop Dis, 2021 Jan;15(1):e0009110.
    PMID: 33493205 DOI: 10.1371/journal.pntd.0009110
    Plasmodium knowlesi is a simian malaria parasite currently recognized as the fifth causative agent of human malaria. Recently, naturally acquired P. cynomolgi infection in humans was also detected in Southeast Asia. The main reservoir of both parasites is the long-tailed and pig-tailed macaques, which are indigenous in this region. Due to increased urbanization and changes in land use, there has been greater proximity and interaction between the long-tailed macaques and the general population in Singapore. As such, this study aims to determine the prevalence of simian malaria parasites in local macaques to assess the risk of zoonosis to the general human population. Screening for the presence of malaria parasites was conducted on blood samples from 660 peridomestic macaques collected between Jan 2008 and Mar 2017, and 379 wild macaques collected between Mar 2009 and Mar 2017, using a Pan-Plasmodium-genus specific PCR. Positive samples were then screened using a simian Plasmodium species-specific nested PCR assay to identify the species of parasites (P. knowlesi, P. coatneyi, P. fieldi, P. cynomolgi, and P. inui) present. All the peridomestic macaques sampled were tested negative for malaria, while 80.5% of the 379 wild macaques were infected. All five simian Plasmodium species were detected; P. cynomolgi being the most prevalent (71.5%), followed by P. knowlesi (47.5%), P. inui (42.0%), P. fieldi (32.5%), and P. coatneyi (28.5%). Co-infection with multiple species of Plasmodium parasites was also observed. The study revealed that Singapore's wild long-tailed macaques are natural hosts of the five simian malaria parasite species, while no malaria was detected in all peridomestic macaques tested. Therefore, the risk of simian malaria transmission to the general human population is concluded to be low. However, this can be better demonstrated with the incrimination of the vectors of simian malaria parasites in Singapore.
  19. 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.
  20. Jeyaprakasam NK, Liew JWK, Low VL, Wan-Sulaiman WY, Vythilingam I
    PLoS Negl Trop Dis, 2020 12;14(12):e0008900.
    PMID: 33382697 DOI: 10.1371/journal.pntd.0008900
    Plasmodium knowlesi, a simian malaria parasite, has been in the limelight since a large focus of human P. knowlesi infection was reported from Sarawak (Malaysian Borneo) in 2004. Although this infection is transmitted across Southeast Asia, the largest number of cases has been reported from Malaysia. The increasing number of knowlesi malaria cases has been attributed to the use of molecular tools for detection, but environmental changes including deforestation likely play a major role by increasing human exposure to vector mosquitoes, which coexist with the macaque host. In addition, with the reduction in human malaria transmission in Southeast Asia, it is possible that human populations are at a greater risk of P. knowlesi infection due to diminishing cross-species immunity. Furthermore, the possibility of increasing exposure of humans to other simian Plasmodium parasites such as Plasmodium cynomolgi and Plasmodium inui should not be ignored. We here review the current status of these parasites in humans, macaques, and mosquitoes to support necessary reorientation of malaria control and elimination in the affected areas.
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