bims-mosdis Biomed News
on Mosquito distribution and disease
Issue of 2020–07–19
23 papers selected by
Richard Halfpenny, Staffordshire University



  1. PLoS One. 2020 ;15(7): e0234959
      The economic and social impacts due to diseases transmitted by mosquitoes in the latest years have been significant. Currently, no specific treatment or commercial vaccine exists for the control and prevention of arboviruses, thereby making entomological characterization fundamental in combating diseases such as dengue, chikungunya, and Zika. The morphological identification of mosquitos includes a visual exam of the samples. It is time consuming and requires adequately trained professionals. Accordingly, the development of a new automated method for realizing mosquito-perception and -classification is becoming increasingly essential. Therefore, in this study, a computational model based on a convolutional neural network (CNN) was developed to extract features from the images of mosquitoes and then classify the species Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus. In addition, the model was trained to detect the mosquitoes of the genus Aedes. To train CNNs to perform the automatic morphological classification of mosquitoes, a dataset, which included 7,561 images of the target mosquitoes and 1,187 images of other insects, was acquired. Various neural networks, such as Xception and DenseNet, were used for developing the automatic-classification model based on images. A structured optimization process of random search and grid search was developed to select the hyperparameters set and increase the accuracy of the model. In addition, strategies to eliminate overfitting were implemented to increase the generalization of the model. The optimized model, during the test phase, obtained the balanced accuracy (BA) of 93.5% in classifying the target mosquitoes and other insects and the BA of 97.3% in detecting the mosquitoes of the genus Aedes in comparison to Culex. The results provide fundamental information for performing the automatic morphological classification of mosquito species. Using a CNN-embedded entomological tool is a valuable and accessible resource for health workers and non-taxonomists for identifying insects that can transmit infectious diseases.
    DOI:  https://doi.org/10.1371/journal.pone.0234959
  2. Parasit Vectors. 2020 Jul 14. 13(1): 352
       BACKGROUND: Aedes aegypti and Culex quinquefasciatus are the main urban vectors of arthropod-borne viruses causing human disease, including dengue, Zika, or West Nile. Although key to disease prevention, urban-mosquito control has met only limited success. Alternative vector-control tactics are therefore being developed and tested, often using entomological endpoints to measure impact. Here, we test one promising alternative and assess how three such endpoints perform at measuring its effects.
    METHODS: We conducted a 16-month, two-arm, cluster-randomized controlled trial (CRCT) of mosquito-disseminated pyriproxyfen (MD-PPF) in central-western Brazil. We used three entomological endpoints: adult-mosquito density as directly measured by active aspiration of adult mosquitoes, and egg-trap-based indices of female Aedes presence (proportion of positive egg-traps) and possibly abundance (number of eggs per egg-trap). Using generalized linear mixed models, we estimated MD-PPF effects on these endpoints while accounting for the non-independence of repeated observations and for intervention-unrelated sources of spatial-temporal variation.
    RESULTS: On average, MD-PPF reduced adult-mosquito density by 66.3% (95% confidence interval, 95% CI: 47.3-78.4%); Cx. quinquefasciatus density fell by 55.5% (95% CI: 21.1-74.8%), and Ae. aegypti density by 60.0% (95% CI: 28.7-77.5%). In contrast, MD-PPF had no measurable effect on either Aedes egg counts or egg-trap positivity, both of which decreased somewhat in the intervention cluster but also in the control cluster. Egg-trap data, therefore, failed to reflect the 60.0% mean reduction of adult Aedes density associated with MD-PPF deployment.
    CONCLUSIONS: Our results suggest that the widely used egg-trap-based monitoring may poorly measure the impact of Aedes control; even if more costly, direct monitoring of the adult mosquito population is likely to provide a much more realistic and informative picture of intervention effects. In our CRCT, MD-PPF reduced adult-mosquito density by 66.3% in a medium-sized, spatially non-isolated, tropical urban neighborhood. Broader-scale trials will be necessary to measure MD-PPF impact on arboviral-disease transmission.
    Keywords:  Cluster randomized controlled trial; Mosquito control; Mosquito-borne diseases; Pyriproxyfen; Vector surveillance
    DOI:  https://doi.org/10.1186/s13071-020-04221-z
  3. J Med Entomol. 2020 Jul 11. pii: tjaa134. [Epub ahead of print]
      Aedes-borne viral diseases such as dengue fever are surging in incidence in recent years. To investigate viral transmission risks, the availability of local transmission parameters is essential. One of the most important factors directly determining infection risk is human-mosquito contact. Yet the contact rate is not often characterized, compared with other risk metrics such as vector density, because of the limited research tool options. In this study, human-mosquito contact was assessed in two study sites in the Southern United States using self-administered standardized survey instruments. The fraction of mosquito bites attributed to important vector species was estimated by human landing sampling. The survey participants reported a significantly higher outdoor mosquito bite exposure than indoor. The reported bite number was positively correlated with outdoor time during at-risk periods. There was also a significant effect of the study site on outdoor bite exposure, possibly due to the differing vector density. Thus, the levels of human-mosquito contact in this study were influenced both by the mosquito density and human behaviors. A dengue virus transmission model demonstrated that the observed difference in the contact rates results in differential virus transmission risks. Our findings highlight the practicality of using surveys to investigate human-mosquito contact in a setting where bite exposure levels differ substantially, and serve as a basis for further evaluations. This study underscores a new avenue that can be used in combination with other field methods to understand how changes in human behavior may influence mosquito bite exposure which drives mosquito-borne virus transmission.
    Keywords:   Aedes aegypti ; Aedes albopictus ; human; SEIR model; dengue virus; mosquito contact
    DOI:  https://doi.org/10.1093/jme/tjaa134
  4. Vector Borne Zoonotic Dis. 2020 Jul 13.
      The pandemic of Zika virus in 2016 and other arboviruses prompted La Rioja Government in Spain to implement an entomological surveillance program of mosquitoes (Diptera; Culicidae) in the region of La Rioja. The morphological identification was supported by genetic analysis using the COI (cytochrome c oxidase subunit I) and the ITS2 (internal transcribed spacer 2) genes. In total, we identified 24 species arranged in 6 genera: Aedes (7 species), Anopheles (4 species), Coquillettidia (1 species), Culex (7 species), Culiseta (4 species), and Uranotaenia (1 species). Aedes sticticus and Aedes geniculatus are newly reported for La Rioja region. In total, 465 COI sequences were analyzed for Culicinae and Anophelinae and 54 ITS2 sequences for Anophelinae; all individuals identified as the same species clustered together in the Neighbor Joining trees. The levels of sequence divergence based on COI ranged between 0% and 2.62%, while the interspecific genetic divergence ranged from 3.05% to 20.07%. Within the genus Culiseta, certain specimens of Culiseta annulata, Culiseta litorea, and Culiseta subochrea were morphologically misidentified due to variation in the main diagnostic characters. The interspecific genetic divergence based on the ITS2 ranged from 0% to 2.98%. An accurate identification of mosquito vectors is the first step to establish a vector surveillance program for preventing pathogen transmission.
    Keywords:  COI; Culicidae; Culiseta; ITS2; genetic diversity; mosquito surveillance
    DOI:  https://doi.org/10.1089/vbz.2020.2662
  5. Parasit Vectors. 2020 Jul 17. 13(1): 353
       BACKGROUND: The wide distribution of Aedes aegypti, the main vector of dengue and yellow fever viruses, currently puts three billion people in the world at risk of infection with these viruses. Continuous transmission of these and other viruses despite aggressive efforts to prevent this emphasizes the need to develop new control strategies. Proposals to control disease transmission based on vector engineering, including both population suppression and population replacement, rely on the development of transgenes under the control of regulatory elements able to drive molecules in a specific tissue, time and strength.
    METHODS: Here we report the characterization of a promoter active in both the female germline and early zygote, derived from the transcription factor bZip1 in the mosquito Ae. aegypti, using transposon-based methods and RT-qPCR.
    RESULTS: We generated seven transgenic lines carrying AabZip1-reporter constructs and observed expression in both the ovary and early embryo. RT-qPCR analysis was performed to evaluate transcript expression patterns for each line, confirming that transgenic expression from the AabZip1 promoter largely recapitulated the endogenous expression pattern, albeit the strength of maternal expression appeared to be strongly influenced by chromosomal position.
    CONCLUSIONS: This study provides a new regulatory sequence that can be useful for generating transgenic lines that can become a tool in vector control strategies.
    Keywords:  Embryo; Germline; Maternal expression; Mosquito; Ovaries; Promoter; Regulatory sequence; Zygote
    DOI:  https://doi.org/10.1186/s13071-020-04216-w
  6. Malar J. 2020 Jul 13. 19(1): 249
       BACKGROUND: New classes of insecticides with novel modes of action, which can provide effective and prolonged control of insecticide-resistant malaria vector populations, are urgently needed for indoor residual spraying. Such insecticides can be included in a rotation plan to manage and prevent further development of resistance in mosquito vectors of malaria. Chlorfenapyr, a novel pyrrole insecticide with a unique mode of action, is being developed as a long-lasting IRS formulation.
    METHODS: The efficacy of several formulations of chlorfenapyr alone and as mixtures with alpha-cypermethrin were evaluated in an experimental hut trial against wild pyrethroid-resistant Anopheles gambiae sensu lato in Cové, Benin, in an attempt to identify the most effective and long-lasting formulations for IRS. The trial lasted 12 months. A comparison was made with alpha-cypermethrin and bendiocarb formulations. CDC bottle bioassays were performed to investigate cross-resistance to chlorfenapyr in the local vector population.
    RESULTS: Mortality rates in World Health Organization (WHO) cylinder bioassays were < 5% with pyrethroids due to high levels of pyrethroid resistance, but > 95% with bendiocarb thus confirming susceptibility to carbamates in the vector population. CDC bottle bioassays showed no cross-resistance between pyrethroids and chlorfenapyr. Overall mortality of free-flying mosquitoes entering the experimental huts over the 12-month trial was 4% with alpha-cypermethrin and 12% with bendiocarb. The chlorfenapyr solo-formulations induced significantly higher levels of mortality (38-46%) compared to the bendiocarb (12% P < 0.001) and to the mixture formulations (18-22%, P < 0.05). The original Sylando 240SC formulation of chlorfenapyr was more efficacious than all other novel chlorfenapyr formulations tested. Bendiocarb induced > 80% mortality in the first month, but this declined sharply to < 20% by the third month while the mortality rates achieved with the chlorfenapyr formulations (38-46%) were persistent lasting 7-10 months. The mixtures induced significantly lower percentage mortality than chlorfenapyr-solo formulations. Wall cone bioassays only showed mortality rates that were consistent with chlorfenapyr IRS treated huts when the exposure time was increased to 2 h.
    CONCLUSION: Indoor residual spraying with chlorfenapyr (Sylando® 240SC) provides moderate but prolonged control of pyrethroid-resistant malaria vectors compared to pyrethroid and bendiocarb IRS. Wall cone bioassays on chlorfenapyr-treated walls required longer exposure times of 2 h than the customary 30 min indicating that WHO guidelines on residual cone bioassays need to be more insecticide-specific.
    Keywords:  Alpha-cypermethrin, Bendiocarb; Anopheles; CDC Bottle bioassays; Chlorfenapyr; Cové; Experimental huts; Indoor residual spraying; Mixtures; Sylando
    DOI:  https://doi.org/10.1186/s12936-020-03325-2
  7. Pathogens. 2020 Jul 12. pii: E564. [Epub ahead of print]9(7):
      The growing expansion of mosquito vectors leads to the emergence of vector-borne diseases in new geographic areas and causes major public health concerns. In the absence of effective preventive treatments against most pathogens transmitted, vector control remains one of the most suitable strategies to prevent mosquito-borne diseases. Insecticide overuse raises mosquito resistance and deleterious impacts on the environment and non-target species. Growing knowledge of mosquito biology has allowed the development of alternative control methods. Following the concept of holobiont, mosquito-microbiota interactions play an important role in mosquito biology. Associated microbiota is known to influence many aspects of mosquito biology such as development, survival, immunity or even vector competence. Mosquito-associated microbiota is composed of bacteria, fungi, protists, viruses and nematodes. While an increasing number of studies have focused on bacteria, other microbial partners like fungi have been largely neglected despite their huge diversity. A better knowledge of mosquito-mycobiota interactions offers new opportunities to develop innovative mosquito control strategies. Here, we review the recent advances concerning the impact of mosquito-associated fungi, and particularly nonpathogenic fungi, on life-history traits (development, survival, reproduction), vector competence and behavior of mosquitoes by focusing on Culex, Aedes and Anopheles species.
    Keywords:  Aedes-Anopheles-Culex spp; mosquito-microbiota interactions; mycobiota
    DOI:  https://doi.org/10.3390/pathogens9070564
  8. J Med Entomol. 2020 Jul 15. pii: tjaa137. [Epub ahead of print]
      Recent results of high-altitude windborne mosquito migration raised questions about the viability of these mosquitoes despite ample evidence that many insect species, including other dipterans, have been known to migrate regularly over tens or hundreds of kilometers on high-altitude winds and retain their viability. To address these concerns, we subjected wild Anopheles gambiae s.l. Giles mosquitoes to a high-altitude survival assay, followed by oviposition (egg laying) and blood feeding assays. Despite carrying out the survival assay under exceptionally harsh conditions that probably provide the lowest survival potential following high altitude flight, a high proportion of the mosquitoes survived for 6- and even 11-h assay durations at 120- to 250-m altitudes. Minimal differences in egg laying success were noted between mosquitoes exposed to high altitude survival assay and those kept near the ground. Similarly, minimal differences were found in the female's ability to take an additional bloodmeal after oviposition between these groups. We conclude that similar to other high-altitude migrating insects, mosquitoes are able to withstand extended high-altitude flight and subsequently reproduce and transmit pathogens by blood feeding on new hosts.
    Keywords:  altitude; disease-vector; egg-laying; long-range dispersal; wind
    DOI:  https://doi.org/10.1093/jme/tjaa137
  9. PLoS Negl Trop Dis. 2020 Jul 17. 14(7): e0008463
      The arbovirus vectors Aedes aegypti (yellow fever mosquito) and Ae. albopictus (Asian tiger mosquito) are both common throughout the Indo-Pacific region, where 70% of global dengue transmission occurs. For Ae. aegypti all Indo-Pacific populations are invasive, having spread from an initial native range of Africa, while for Ae. albopictus the Indo-Pacific includes invasive populations and those from the native range: putatively, India to Japan to Southeast Asia. This study analyses the population genomics of 480 of these mosquitoes sampled from 27 locations in the Indo-Pacific. We investigated patterns of genome-wide genetic differentiation to compare pathways of invasion and ongoing gene flow in both species, and to compare invasive and native-range populations of Ae. albopictus. We also tested landscape genomic hypotheses that genetic differentiation would increase with geographical distance and be lower between locations with high connectivity to human transportation routes, the primary means of dispersal at these scales. We found that genetic distances were generally higher in Ae. aegypti, with Pacific populations the most highly differentiated. The most differentiated Ae. albopictus populations were in Vanuatu, Indonesia and Sri Lanka, the latter two representing potential native-range populations and potential cryptic subspeciation respectively. Genetic distances in Ae. aegypti increased with geographical distance, while in Ae. albopictus they decreased with higher connectivity to human transportation routes. Contrary to the situation in Ae. aegypti, we found evidence of long-distance Ae. albopictus colonisation events, including colonisation of Mauritius from East Asia and of Fiji from Southeast Asia. These direct genomic comparisons indicate likely differences in dispersal ecology in these species, despite their broadly sympatric distributions and similar use of human transport to disperse. Our findings will assist biosecurity operations to trace the source of invasive material and for biocontrol operations that benefit from matching genetic backgrounds of released and local populations.
    DOI:  https://doi.org/10.1371/journal.pntd.0008463
  10. MMWR Morb Mortal Wkly Rep. 2020 Jul 17. 69(28): 938-940
      Mosquitoborne disease outbreaks occur every year in the United States from one or more of the arboviral diseases dengue, West Nile, LaCrosse, Eastern equine encephalitis, and Zika (1). Public opinion communicated through traditional and social media and the Internet, competing public health and resource priorities, and local conditions can impede the ability of vector control organizations to prevent and respond to outbreaks of mosquitoborne disease. The Environmental Protection Agency (EPA) and CDC performed a coordinated review of the concerns and challenges associated with continuation of mosquito surveillance and control during public health emergencies and disasters. This report highlights the first joint recommendation from EPA and CDC. Mosquito surveillance and control should be maintained by state and local mosquito control organizations to the extent that local conditions and resources will allow during public health emergencies and natural disasters. Integrated pest management (IPM) is the best approach for mosquito control (2). IPM uses a combination of methods, including both physical and chemical means of control (3). For chemical means of control, CDC and EPA recommend the use of larvicides and adulticides following the EPA label. It is imperative that public health recommendations be followed to ensure the safety of the pesticide applicator and the public.
    DOI:  https://doi.org/10.15585/mmwr.mm6928a6
  11. J Vector Borne Dis. 2019 Jul-Sep;56(3):56(3): 207-211
       BACKGROUND & OBJECTIVES: The emergence and spread of insecticide resistance in African malaria vectors raise concerns over the control of malaria disease. Therefore, the implementation of better control strategies need a thorough understanding of the effects and mechanisms of resistance on vector adaptation capacities. We studied the effects of insecticide resistance on the reproductive potential of two laboratory sub-strains of the malaria vector Anopheles coluzzii characterised by phenotypic resistance/susceptibility to DDT.
    METHODS: The two sub-strains were selected from a laboratory strain of An. coluzzii using WHO test tubes. For each sub-strain, the number of produced and hatched eggs, developmental time, mosquito stages mortality, sex ratio and insemination rates after dissection of spermathecae were compared as measures of reproductive potential.
    RESULTS: Overall, the susceptible sub-strain produced higher but not significant mean numbers of eggs. However, the mean numbers of hatched eggs, larvae, pupae and adults were significantly lower than those of the resistant substrain. The mean time from egg-hatching to adult-emergence, egg-flooding to hatching, I instar to pupae and pupae to adult were similar between the two sub-strains. The mortality rates at the pupal stage were significantly different between the two sub-strains. Of the dissected spermathecae, 85.1% of the females from the resistant sub-strain were fertilized compared to 66.1% of the females from the susceptible sub-strain (p <0.0001). The resistant sub-strain produced more females in comparison to the susceptible sub-strain (respective mean sex ratio 1.37 vs 1.03, p = 0.01).
    INTERPRETATION & CONCLUSION: The results show differential life history traits between the two sub-strains of the malaria vector An. coluzzii, particularly fertility, insemination rate and sex ratio. They may have varied implications for insecticide resistance spread, monitoring and management; and hence underscore the need of further investigations before any generalization.
    Keywords:  Anopheles coluzzii; Senegal; insecticide resistance; reproductive potential
    DOI:  https://doi.org/10.4103/0972-9062.289401
  12. J Vector Borne Dis. 2019 Jul-Sep;56(3):56(3): 200-206
       BACKGROUND & OBJECTIVES: Although malaria is eliminated from Sri Lanka, there is a possible risk of spread from infected persons coming from malaria endemic countries. The presence of major and potential vectors in several parts of the country along with drug resistance, necessitates the identification of effective and novel control methods. The present study focused on identifying effective biological control agents for anopheline larvae using carnivorous copepods under laboratory and field conditions to prevent re-introduction of malaria in the country.
    METHODS: Three copepod species, namely Mesocyclops scrassus, Cyclops varicans and C. languides collected from different areas in the country were cultured by adding supplementary food, and their predatory efficacy was evaluated under laboratory and field conditions.
    RESULTS: Significant variation (p <0.05) was observed in predation rates of studied copepod species. The species M. scrassus showed the highest predacious efficiency, and consumed the highest number of anopheline larvae under laboratory and field conditions. Further, M. scrassus had higher survival rate than C. varicans and C. languides.
    INTERPRETATION & CONCLUSION: The results of the study suggest that the predatory copepod M. scrassus can be used as a bio-control agent for the control of Anopheles mosquitoes to prevent re-emergence of malaria in the country. Additional research is suggested to identify naturally available copepod species and their predatory efficacy.
    Keywords:  Anopheles; Sri Lanka; biological control; copepods; malaria vectors; predatory efficacy
    DOI:  https://doi.org/10.4103/0972-9062.289393
  13. Front Med (Lausanne). 2020 ;7 275
      Continuous climate changes associated with the disorderly occupation of urban areas have exposed Latin American populations to the emergence and reemergence of arboviruses transmitted by Aedes aegypti. The magnitude of the financial and political problems these epidemics may bring to the future of developing countries is still ignored. Due to the lack of effective antiviral drugs and vaccines against arboviruses, the primary measure for preventing or reducing the transmission of diseases depends entirely on the control of vectors or the interruption of human-vector contact. In Brazil the first attempt to control A. aegypti took place in 1902 by eliminating artificial sites of eproduction. Other strategies, such as the use of oviposition traps and chemical control with dichlorodiphenyltrichlorethane and pyrethroids, were successful, but only for a limited time. More recently, biotechnical approaches, such as the release of transgenics or sterile mosquitoes and the, development of transmission blocking vaccines, are being applied to try to control the A. aegypti population and/or arbovirus transmission. Endemic countries spend about twice as much to treat patients as they do on the prevention of mosquito-transmitted diseases. The result of this strategy is an explosive outbreak of arboviruses cases. This review summarizes the social impacts caused by A. aegypti-transmitted diseases, mainly from a biotechnological perspective in vector control aimed at protecting Latin American populations against arboviruses.
    Keywords:  Aedes aegypti; Latin America; arboviruses; vaccines; vector control
    DOI:  https://doi.org/10.3389/fmed.2020.00275
  14. Malar J. 2020 Jul 11. 19(1): 242
       BACKGROUND: Ghana has been implementing the indoor residual spraying (IRS) of insecticides since 2006, focusing operations in the north. Insecticide resistance concerns prompted a switch from pyrethroids to organophosphates, beginning gradually in 2011 and switching fully to the micro-encapsulated formulation of pirimiphosmethyl (PM CS), Actellic® 300CS, a third-generation indoor residual spraying (3GIRS) product, by 2014. Entomological surveillance studies have shown IRS to be a highly effective malaria control tool, but epidemiological evidence is needed as well. Countrywide prevalence surveys have shown that malaria parasite prevalence in children under 5 years of age in Northern, Upper East, and Upper West Regions had declined to less than 40% in each region by 2016. Similarly, malaria deaths in children under 5 years of age have also been declining nationally since 2009. Although IRS is suspected to have contributed to this decline, stronger evidence is needed to link the IRS interventions to the epidemiological impact.
    METHODS: To assess the epidemiological impact of Ghana's IRS programmatic activities, a retrospective, observational analysis using routine epidemiological data was conducted to compare malaria incidence rates from IRS and non-IRS districts in Northern, Upper East, and Upper West Regions. Routine epidemiological data consisted of passive malaria case surveillance data reported in the District Health Information System 2 (DHIS2); with cases representing patients with suspected malaria who had sought care in the public health system and had received a confirmatory diagnosis with a positive malaria RDT result. Final routine data were extracted in September 2018. All districts that had received IRS were included in the analysis and compared to all non-IRS districts within the same region. In the Northern Region, only PMI districts were included in the analysis, as they had similar historical data.
    RESULTS: District-level analysis from Northern Region from 2015 to 2017 of the aggregate malaria incidence reported from IRS districts relative to non-IRS comparator districts showed 39%, 26%, and 58% fewer confirmed malaria cases reported from IRS districts in 2015, 2016, and 2017, respectively. This translates to approximately 257,000 fewer cases than expected over the three years. In Upper East Region, the effect on reported malaria cases of withdrawing IRS from the region was striking; after spray operations were suspended in 2015, incidence increased an average of 485% per district (95% confidence interval: 330% to 640%) compared to 2014.
    CONCLUSIONS: The current observational analysis results are in line with the entomological studies in demonstrating the positive contribution of IRS with a 3GIRS product to malaria control programmes in northern Ghana and the value of using routine surveillance and implementation data to rapidly assess the impact of vector control interventions in operational settings, even in complex implementation environments.
    Keywords:  3GIRS; Actellic; IRS; Indoor residual spraying; Malaria; PM CS; Pirimiphosmethyl; Third-generation IRS; Vector control
    DOI:  https://doi.org/10.1186/s12936-020-03318-1
  15. Parasit Vectors. 2020 Jul 13. 13(1): 349
       BACKGROUND: The ability of blood-feeding arthropods to successfully acquire and transmit pathogens of medical and veterinary importance has been shown to be interfered with, or enhanced by, the arthropod's native microbiome. Mosquitoes transmit viruses, protozoan and filarial nematodes, the majority of which contribute to the 17% of infectious disease cases worldwide. Dirofilaria immitis, a mosquito-transmitted filarial nematodes of dogs and cats, is vectored by several mosquito species including Aedes aegypti.
    METHODS: In this study, we investigated the impact of D. immitis colonization on the microbiome of laboratory reared female Ae. aegypti. Metagenomic analysis of the V3-V4 variable region of the microbial 16S RNA gene was used for identification of the microbial differences down to species level.
    RESULTS: We generated a total of 1068 OTUs representing 16 phyla, 181 genera and 271 bacterial species. Overall, in order of abundance, Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes were the most represented phylum with D. immitis-infected mosquitoes having more of Proteobacteria (71%) than uninfected mosquitoes (56.9%). An interesting finding in this study is the detection of Klebsiella oxytoca in relatively similar abundance in infected and uninfected mosquitoes, suggesting a possible endosymbiotic relationship, and has been previously shown to indirectly compete for nutrients with fungi on domestic housefly eggs and larvae. While D. immitis colonization has no effect on the overall species richness, we identified significant differences in the composition of selected bacterial genera and phyla between the two groups. We also reported distinct compositional and phylogenetic differences in the individual bacterial species when commonly identified bacteria were compared.
    CONCLUSIONS: To the best of our knowledge, this is the first study to understand the impact of a filarial infection on the microbiome of its mosquito vector. Further studies are required to identify bacteria species that could play an important role in the mosquito biology. While the microbiome composition of Ae. aegypti mosquito have been previously reported, our study shows that in an effort to establish itself, a filarial nematode modifies and alters the overall microbial diversity within its mosquito host.
    Keywords:  Aedes aegypti; Dirofilaria immitis; Dog heartworm; Metagenome; Microbiome; Mosquitoes
    DOI:  https://doi.org/10.1186/s13071-020-04218-8
  16. Parasitol Res. 2020 Jul 16.
      Before the background of increasingly frequent outbreaks and cases of mosquito-borne diseases in various European countries, Germany recently realised the necessity of updating decade-old data on the occurrence and spatiotemporal distribution of culicid species. Starting in 2011, a mosquito monitoring programme was therefore launched with adult and immature mosquito stages being collected at numerous sites all over Germany both actively by trapping, netting, aspirating and dipping, and passively by the citizen science project 'Mueckenatlas'. Until the end of 2019, about 516,000 mosquito specimens were analysed, with 52 (probably 53) species belonging to seven genera found, including several species not reported for decades due to being extremely rare (Aedes refiki, Anopheles algeriensis, Culex martinii) or local (Culiseta alaskaensis, Cs. glaphyroptera, Cs. ochroptera). In addition to 43 (probably 44 including Cs. subochrea) out of 46 species previously described for Germany, nine species were collected that had never been documented before. These consisted of five species recently established (Ae. albopictus, Ae. japonicus, Ae. koreicus, An. petragnani, Cs. longiareolata), three species probably introduced on one single occasion only and not established (Ae. aegypti, Ae. berlandi, Ae. pulcritarsis), and a newly described cryptic species of the Anopheles maculipennis complex (An. daciae) that had probably always been present but not been differentiated from its siblings. Two species formerly listed for Germany could not be documented (Ae. cyprius, Ae. nigrinus), while presence is likely for another species (Cs. subochrea), which could not be demonstrated in the monitoring programme as it can neither morphologically nor genetically be reliably distinguished from a closely related species (Cs. annulata) in the female sex. While Cs. annulata males were collected in the present programme, this was not the case with Cs. subochrea. In summary, although some species regarded endemic could not be found during the last 9 years, the number of culicid species that must be considered firmly established in Germany has increased to 51 (assuming Cs. subochrea and Ae. nigrinus are still present) due to several newly emerged ones but also to one species (Ae. cyprius) that must be considered extinct after almost a century without documentation. Most likely, introduction and establishment of the new species are a consequence of globalisation and climate warming, as three of them are native to Asia (Ae. albopictus, Ae. japonicus, Ae. koreicus) and three (Ae. albopictus, An. petragnani, Cs. longiareolata) are relatively thermophilic. Another thermophilic species, Uranotaenia unguiculata, which had been described for southwestern Germany in 1994 and had since been found only at the very site of its first detection, was recently documented at additional localities in the northeastern part of the country. As several mosquito species found in Germany are serious pests or potential vectors of disease agents and should be kept under permanent observation or even be controlled immediately on emergence, the German mosquito monitoring programme has recently been institutionalised and perpetuated.
    Keywords:  Climate warming; Culicidae; Germany; Globalisation; Invasive species; Inventory; Mosquito monitoring
    DOI:  https://doi.org/10.1007/s00436-020-06775-4
  17. Front Microbiol. 2020 ;11 1297
      Chikungunya virus (CHIKV) is an arthropod-borne virus (arbovirus) of epidemic concern, transmitted by Aedes ssp. mosquitoes, and is the etiologic agent of a febrile and incapacitating arthritogenic illness responsible for millions of human cases worldwide. After major outbreaks starting in 2004, CHIKV spread to subtropical areas and western hemisphere coming from sub-Saharan Africa, South East Asia, and the Indian subcontinent. Even though CHIKV disease is self-limiting and non-lethal, more than 30% of the infected individuals will develop chronic disease with persistent severe joint pain, tenosynovitis, and incapacitating polyarthralgia that can last for months to years, negatively impacting an individual's quality of life and socioeconomic productivity. The lack of specific drugs or licensed vaccines to treat or prevent CHIKV disease associated with the global presence of the mosquito vector in tropical and temperate areas, representing a possibility for CHIKV to continually spread to different territories, make this virus an agent of public health burden. In South America, where Dengue virus is endemic and Zika virus was recently introduced, the impact of the expansion of CHIKV infections, and co-infection with other arboviruses, still needs to be estimated. In Brazil, the recent spread of the East/Central/South Africa (ECSA) and Asian genotypes of CHIKV was accompanied by a high morbidity rate and acute cases of abnormal disease presentation and severe neuropathies, which is an atypical outcome for this infection. In this review, we will discuss what is currently known about CHIKV epidemics, clinical manifestations of the human disease, the basic concepts and recent findings in the mechanisms underlying virus-host interaction, and CHIKV-induced chronic disease for both in vitro and in vivo models of infection. We aim to stimulate scientific debate on how the characterization of replication, host-cell interactions, and the pathogenic potential of the new epidemic viral strains can contribute as potential developments in the virology field and shed light on strategies for disease control.
    Keywords:  Chikungunya virus; emergent arbovirus; epidemiology; pathogenesis; virus–cell interaction
    DOI:  https://doi.org/10.3389/fmicb.2020.01297
  18. J Vector Borne Dis. 2019 Jul-Sep;56(3):56(3): 179-188
      Sri Lanka is a country that has long suffered from epidemics of malaria. In this historical context, it is remarkable that in 2016 the Indian Ocean island nation was able to officially celebrate the elimination of this parasitic disease of major public health importance. The most devastating outbreak recorded in Sri Lanka was during 1934-35, when close to 80,000 human deaths were reported. Indoor residual spraying with the insecticides, DDT and malathion commenced in 1947 and was successful in causing a rapid decline in malaria incidence. However, poor vector control measures, resistance of mosquitoes to these insecticides and resistance of blood-stage Plasmodium parasites to the prevailing drugs used are considered the principal reasons for the occurrence of subsequent outbreaks. Despite this, Sri Lanka achieved the significant milestone of zero locally transmitted malaria cases in October 2012 and zero recorded deaths since 2007. Vector surveillance, parasitological examination, and clinical case management were collective effective activities that most likely led to elimination of malaria. Yet, there remains a high risk of reintroduction due to imported cases and an enduring vulnerability to vector transmission. In order to prevent re-establishment of malaria, continued financial support, sustained surveillance for vector species present in Sri Lanka and effective control of imported cases through rapid detection and early diagnosis are all required. In addition to these immediate practical priorities, further studies on vector biology and genetic variations that affect vectorial capacity would help to shed light on how to avoid reintroduction. This review affords an insight into the determinants of past malaria epidemics, strategies deployed to achieve and maintain the current status of elimination, lessons learnt from this success and plans to avoid resurgence of infection.
    Keywords:  Elimination; Plasmodium; Sri Lanka; importation; malaria; vector
    DOI:  https://doi.org/10.4103/0972-9062.289390
  19. Malar J. 2020 Jul 17. 19(1): 258
       BACKGROUND: While Iran is on the path to eliminating malaria, the disease with 4.9 million estimated cases and 9300 estimated deaths in 2018 remains a serious health problem in the World Health Organization (WHO) Eastern Mediterranean Region. Anopheles stephensi is the main malaria vector in Iran and its range extends from Iraq to western China. Recently, the vector invaded new territories in Sri Lanka and countries in the Horn of Africa. Insecticide resistance in An. stephensi is a potential issue in controlling the spread of this vector.
    METHODS: Data were collated from national and international databases, including PubMed, Google Scholar, Scopus, ScienceDirect, SID, and IranMedex using appropriate search terms.
    RESULTS: Indoor residual spaying (IRS) with DDT was piloted in Iran in 1945 and subsequently used in the malaria eradication programme. Resistance to DDT in An. stephensi was detected in Iran, Iraq, Pakistan, and Saudi Arabia in the late 1960s. Malathion was used for malaria control in Iran in 1967, then propoxur in 1978, followed by pirimiphos-methyl from 1992 to 1994. The pyrethroid insecticide lambda-cyhalothrin was used from 1994 to 2003 followed by deltamethrin IRS and long-lasting insecticidal nets (LLINs). Some of these insecticides with the same sequence were used in other malaria-endemic countries of the region. Pyrethroid resistance was detected in An. stephensi in Afghanistan in 2010, in 2011 in India and in 2012 in Iran. The newly invaded population of An. stephensi in Ethiopia was resistant to insecticides of all four major insecticide classes. Different mechanisms of insecticide resistance, including metabolic and insecticide target site insensitivity, have been developed in An. stephensi. Resistance to DDT was initially glutathione S-transferase based. Target site knockdown resistance was later selected by pyrethroids. Esterases and altered acetylcholinesterase are the underlying cause of organophosphate resistance and cytochrome p450s were involved in pyrethroid metabolic resistance.
    CONCLUSIONS: Anopheles stephensi is a major malaria vector in Iran and many countries in the region and beyond. The species is leading in terms of development of insecticide resistance as well as developing a variety of resistance mechanisms. Knowledge of the evolution of insecticide resistance and their underlying mechanisms, in particular, are important to Iran, considering the final steps the country is taking towards malaria elimination, but also to other countries in the region for their battle against malaria. This systematic review may also be of value to countries and territories newly invaded by this species, especially in the Horn of Africa, where the malaria situation is already dire.
    Keywords:  Anopheles stephensi; Eastern Mediterranean Region; Iran; Mechanisms; Resistance
    DOI:  https://doi.org/10.1186/s12936-020-03335-0
  20. Infect Genet Evol. 2020 Jul 12. pii: S1567-1348(20)30289-6. [Epub ahead of print] 104458
      Malaria continues to be one of the top infectious agents contributing to morbidity and mortality in sub-Saharan Africa. Annually, Botswana accounts only for a small proportion of cases (<<1%). Despite significantly reduced incidence rate, the country still experiences sporadic outbreaks that hamper the goal of malaria elimination. This review evaluated previous and current biological factors that impact malaria in Botswana, specifically focussing on the vectors, the parasite and the host. This was accomplished via a literature review evaluating these variables in Botswana. Current literature suggests that Anopheles arabiensis is the main malaria vector in the country. Several other potential vectors have been found widely distributed throughout Botswana in high numbers, yet remain largely unstudied with regards to their contribution to the country's malaria burden. We also report the most up to date list of all Anopheles species that have been found in Botswana. Plasmodium falciparum is responsible for the vast majority of symptomatic malaria in the country and some drug resistance markers have been documented for this species. Plasmodium vivax has been reported in asymptomatic subjects, even though a large proportion of the Botswana population appears to be Duffy antigen negative. Very little is known about the true distribution of P. vivax and no point of care testing infrastructure for this species exists in Botswana, making it difficult to tailor treatment to address possible recrudescence or relapse. Due to a genetically diverse population with a substantial Khoisan contribution into the Bantu genetic background, several phenotypes that potentially impact prevalence and severity of malaria exist within the country. These include sickle cell trait, Glucose-6-Phosphate Dehydrogenase deficiency, and Duffy negativity. This review highlights the information that currently exists on malaria in Botswana. It also postulates that a comprehensive understanding of these aforementioned biological factors may help to explain malaria persistence in Botswana.
    Keywords:  Anopheles; Botswana; Human genetics; Malaria; Plasmodium; Vectors
    DOI:  https://doi.org/10.1016/j.meegid.2020.104458
  21. Med Vet Entomol. 2020 Jul 15.
      This study aimed to evaluate the effects of egg dormancy times on susceptibility of larvae of the floodwater mosquito Aedes albifasciatus (Diptera: Culicidae) to parasitism by their natural enemy Strelkovimermis spiculatus (Nematoda: Mermithidae) and on their life history traits. Aedes albifasciatus eggs stored for 2, 4, 6, 8 and 10 months were hatched, and the larvae either exposed to S. spiculatus (treatment group) or not exposed (control group). Egg dormancy time had a negative effect on the retention of parasites, but no effect on the prevalence and intensity of parasitism or the melanization of nematodes. The survival to adulthood of control individuals decreased as dormancy time increased, whereas that of exposed individuals that remained uninfected was constant and low. A trend towards increasing development times with longer dormancy times was detected in the control group, but not in the exposed noninfected group. The results suggest nonconsumptive effects of parasites in exposed but not infected larvae from eggs with short dormancy times. In contrast, the relatively low fitness of larvae from eggs with long dormancy times regardless of their contact with the nematodes may be the result of the nutritional deprivation during the egg stage.
    Keywords:  Culicidae; Strelkovimermis spiculatus; dormant; egg age; floodwater mosquito ecology; mermithidae; natural host
    DOI:  https://doi.org/10.1111/mve.12460
  22. Mem Inst Oswaldo Cruz. 2020 ;pii: S0074-02762020000100326. [Epub ahead of print]115 e200046
      BACKGROUND Fluctuations in climate have been associated with variations in mosquito abundance. OBJECTIVES To analyse the influence of precipitation, temperature, solar radiation, wind speed and humidity on the oviposition dynamics of Aedes aegypti in three distinct environmental areas (Brasília Teimosa, Morro da Conceição/Alto José do Pinho and Dois Irmãos/Pintos) of the city of Recife and the Fernando de Noronha Archipelago northeastern Brazil. METHODS Time series study using a database of studies previously carried out in the areas. The eggs were collected using spatially distributed geo-referenced sentinel ovitraps (S-OVTs). Meteorological satellite data were obtained from the IRI climate data library. The association between meteorological variables and egg abundance was analysed using autoregressive models. FINDINGS Precipitation was positively associated with egg abundance in three of the four study areas with a lag of one month. Higher humidity (β = 45.7; 95% CI: 26.3 - 65.0) and lower wind speed (β = -125.2; 95% CI: -198.8 - -51.6) were associated with the average number of eggs in the hill area. MAIN CONCLUSIONS The effect of climate variables on oviposition varied according to local environmental conditions. Precipitation was a main predictor of egg abundance in the study settings.
    DOI:  https://doi.org/10.1590/0074-02760200046
  23. Infect Dis Poverty. 2020 Jul 16. 9(1): 95
       BACKGROUND: Disease surveillance systems are essential for effective disease intervention and control by monitoring disease prevalence as time series. To evaluate the severity of an epidemic, statistical methods are widely used to forecast the trend, seasonality, and the possible number of infections of a disease. However, most statistical methods are limited in revealing the underlying dynamics of disease transmission, which may be affected by various impact factors, such as environmental, meteorological, and physiological factors. In this study, we focus on investigating malaria transmission dynamics based on time series data.
    METHODS: A data-driven nonlinear stochastic model is proposed to infer and predict the dynamics of malaria transmission based on the time series of prevalence data. Specifically, the dynamics of malaria transmission is modeled based on the notion of vectorial capacity (VCAP) and entomological inoculation rate (EIR). A particle Markov chain Monte Carlo (PMCMC) method is employed to estimate the model parameters. Accordingly, a one-step-ahead prediction method is proposed to project the number of future malaria infections. Finally, two case studies are carried out on the inference and prediction of Plasmodium vivax transmission in Tengchong and Longling, Yunnan province, China.
    RESULTS: The results show that the trained data-driven stochastic model can well fit the historical time series of P. vivax prevalence data in both counties from 2007 to 2010. Moreover, with well-trained model parameters, the proposed one-step-ahead prediction method can achieve better performances than that of the seasonal autoregressive integrated moving average model with respect to predicting the number of future malaria infections.
    CONCLUSIONS: By involving dynamically changing impact factors, the proposed data-driven model together with the PMCMC method can successfully (i) depict the dynamics of malaria transmission, and (ii) achieve accurate one-step-ahead prediction about malaria infections. Such a data-driven method has the potential to investigate malaria transmission dynamics in other malaria-endemic countries/regions.
    DOI:  https://doi.org/10.1186/s40249-020-00696-1