Commenced in January 2007
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Edition: International
Paper Count: 31

Search results for: Aedes aegypti

31 Representative Concentration Pathways Approach on Wolbachia Controlling Dengue Virus in Aedes aegypti

Authors: Ida Bagus Mandhara Brasika, I Dewa Gde Sathya Deva


Wolbachia is recently developed as the natural enemy of Dengue virus (DENV). It inhibits the replication of DENV in Aedes aegypti. Both DENV and its vector, Aedes aegypty, are sensitive to climate factor especially temperature. The changing of climate has a direct impact on temperature which means changing the vector transmission. Temperature has been known to effect Wolbachia density as it has an ideal temperature to grow. Some scenarios, which are known as Representative Concentration Pathways (RCPs), have been developed by Intergovernmental Panel on Climate Change (IPCC) to predict the future climate based on greenhouse gases concentration. These scenarios are applied to mitigate the future change of Aedes aegypti migration and how Wolbachia could control the virus. The prediction will determine the schemes to release Wolbachia-injected Aedes aegypti to reduce DENV transmission.

Keywords: Aedes aegypti, climate change, dengue virus, Intergovernmental Panel on Climate Change, representative concentration pathways, Wolbachia

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30 Age-Stage, Two-Sex Life Table Characteristics of Aedes albopictus (Skuse) and Aedes aegypti (Linnaeus)) (Diptera: Culicidae) in Penang Island, Malaysia

Authors: A. H. Maimusa, A. Abu Hassan, Nur Faeza A. Kassim


In this study, we report on the main life table developmental attributes of laboratory colonies of wild strains Ae. albopictus and Ae. aegypti. The raw life history data of the two species were analyzed and compared based on the age-stage and two-sex life table. The total pre-adult development times were 9.47 days (Ae. albopictus) and 8.76 days (Ae. aegypti). The adult pre-oviposition periods (APOP) was 1.61 day for Ae. albopictus and 2.02 for Ae. aegypti. The total pre-oviposition period (TPOP) of Ae. albopictus is significantly longer (11.66 days) than (10.75 days) for Ae. aegypti. The mean intrinsic rate of increase (r) was 0.124 days (Ae. albopictus) and 1.151 days (Ae. aegypti) while the mean finite rate of increase (λ) was 1.13 day (Ae. albopictus) and (1.16 d) (Ae. aegypti). The net reproductive rates (Ro) were 8.10 and 10.75 for Ae. albopictus and Ae. aegypti, respectively. The mean generation time (T) for Ae. albopictus and Ae. aegypti, were 16.81 days and 15.77 days respectively. The mean development time for each stage insignificantly correlated with temperature (r = -0.208, p > 0.05) and (r = -0.312, p > 0.05) for Ae. albopictus and Ae. aegypti respectively. The life expectancy was 19.01 and 19.94 days for Ae. albopictus and Ae. aegypti respectively. Mortality occurred mostly during the adult stage and ranged between 0.01 and 0.07%. The population parameters suggest that Ae. albopictus and Ae. aegypti populations are r-strategist characterized by a high r, a large Ro, and short T. This kind of information is crucial in understanding mosquito population dynamics in disease transmission and control.

Keywords: Ae. aegypti, Ae. albopictus, age-stage, life table, two-sex

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29 Bioefficacy of Ocimum sanctum on Survival, Development and Reproduction of Dengue Vector Aedes aegypti L. (Diptera: Culicidae)

Authors: Mohd Shazad, K. K. Gupta


Vector borne diseases are a serious global concern. Aedes aegypti, the primary vector for viruses that cause dengue fever, dengue haemorrhagic fever, chikungunya and yellow fever is widespread over large areas of the tropics and subtropics. In last decade, diseases transmitted by Aedes aegypti are of serious concern. In past decade, number of cases of dengue fever, dengue hemorrhagic fever, and chikungunya has increased multifold. Present research work focused on impact of ethanol extract of Ocimum sanctum on dengue vector Aedes aegypti. 0-24 hr. old fourth instar larvae of lab-bred population of Aedes aegypti were exposed to ethanol leaf extract of Ocimum with concentrations ranging from 50 ppm to 400 ppm. Survival and development and the treated larvae and reproductive behaviour of the adults emerged from the treated larvae was evaluated. Our results indicated larvicidal potential of the leaf ethanol extract. The influence of the extract was dose dependent. 77.2% mortality was observed in the larvae exposed to 400 ppm for 24 hr. Treatment at lower concentrations revealed delayed toxicity. The larvae survived after treatment showed severe developmental anomalies. Consequently, there was the significant increase in duration of fourth instar larva. The L4 treated with 400-ppm extract moulted after 4.6 days; this was in sharp contrast to control where the larval period of the fourth instar lasts three days. The treated fourth instar larvae in many cases transformed into larva-pupa intermediates with the combination of larva, pupa characters. The larva-pupa intermediates had reduced life span and failed to moult successfully. The adults emerged from the larvae treated with lower doses had reduced reproductive potential. The females exhibited longer preoviposition period, reduced oviposition rate, abnormal oviposition behaviour and decreased fertility. Our studies indicated the possibility of the presence of JH mimic or JH analogue in the leaf ethanol extract of Ocimum. The present research work explored the potentials of Ocimum sanctum, also known as the queen of herbs, in integrated vector management programme of Aedes aegypti, which is a serious threat to human health.

Keywords: Aedes aegypti, development, mortality, Ocimum sanctum reproduction

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28 Dengue Virus Infection Rate in Mosquitoes Collected in Thailand Related to Environmental Factors

Authors: Chanya Jetsukontorn


Dengue hemorrhagic fever is the most important Mosquito-borne disease and the major public health problem in Thailand. The most important vector is Aedes aegypti. Environmental factors such as temperature, relative humidity, and biting rate affect dengue virus infection. The most effective measure for prevention is controlling of vector mosquitoes. In addition, surveillance of field-caught mosquitoes is imperative for determining the natural vector and can provide an early warning sign at risk of transmission in an area. In this study, Aedes aegypti mosquitoes were collected in Amphur Muang, Phetchabun Province, Thailand. The mosquitoes were collected in the rainy season and the dry season both indoor and outdoor. During mosquito’s collection, the data of environmental factors such as temperature, humidity and breeding sites were observed and recorded. After identified to species, mosquitoes were pooled according to genus/species, and sampling location. Pools consisted of a maximum of 10 Aedes mosquitoes. 70 pools of 675 Aedes aegypti were screened with RT-PCR for flaviviruses. To confirm individual infection for determining True infection rate, individual mosquitoes which gave positive results of flavivirus detection were tested for dengue virus by RT-PCR. The infection rate was 5.93% (4 positive individuals from 675 mosquitoes). The probability to detect dengue virus in mosquitoes at the neighbour’s houses was 1.25 times, especially where distances between neighboring houses and patient’s houses were less than 50 meters. The relative humidity in dengue-infected villages with dengue-infected mosquitoes was significantly higher than villages that free from dengue-infected mosquitoes. Indoor biting rate of Aedes aegypti was 14.87 times higher than outdoor, and biting times of 09.00-10.00, 10.00-11.00, 11.00-12.00 yielded 1.77, 1.46, 0.68mosquitoes/man-hour, respectively. These findings confirm environmental factors were related to Dengue infection in Thailand. Data obtained from this study will be useful for the prevention and control of the diseases.

Keywords: Aedes aegypti, Dengue virus, environmental factors, one health, PCR

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27 In vitro Control of Aedes aegypti Larvae Using Beauveria bassiana

Authors: R. O. B. Bitencourt, F. S. Farias, M. C. Freitas, C. J. R. Balduino, E.S. Mesquita, A. R. C. Corval, P. S. Gôlo, E. G. Pontes, V. R. E. P. Bittencourt, I. C. Angelo


Aedes aegypti larval survival rate was assessed after exposure to blastopores or conidia (mineral oil-in-water formulation or aqueous suspension) of Beauveria bassiana CG 479 propagules (blastospores or conidia). Here, mineral oil was used in the fungal formulation to control Aedes aegypti larvae. 1%, 0.5% or 0.1% mineral oil-in-water solutions were used to evaluate mineral oil toxicity for mosquito larvae. In the oil toxicity test, 0.1% mineral oil solution reduced only 4.5% larval survival; accordingly, this concentration was chosen for fungal oil-in-water formulations. Aqueous suspensions were prepared using 0.01% Tween 80® in sterile dechlorinated water. A. aegypti larvae (L2) were exposed in aqueous suspensions or mineral oil-in-water fungal formulations at 1×107 propagules mL-1; the survival rate (assessed daily, for 7 days) and the median survival time (S50) were calculated. Seven days after the treatment, mosquito larvae survival rates were 8.56%, 16.22%, 58%, and 42.56% after exposure to oil-in-water blastospores, oil-in-water conidia, blastospores aqueous suspension and conidia aqueous suspension (respectively). Larvae exposed to 0.01% Tween 80® had 100% survival rate and the ones treated with 0.1% mineral oil-in-water had 95.11% survival rate. Larvae treated with conidia (regardless the presence of oil) or treated with blastospores formulation had survival median time (S50) ranging from one to two days. S50 was not determined (ND) when larvae were exposed to blastospores aqueous suspension, 0.01% Tween 80® (aqueous control) or 0.1% mineral oil-in-water formulation (oil control). B. bassiana conidia and blastospores (mineral oil-in-water formulated or suspended in water) had potential to control A. aegypti mosquito larvae, despite mineral oil-in-water formulation yielded better results in comparison to aqueous suspensions. Here, B. bassiana CG 479 isolate is suggested as a potential biocontrol agent of A. aegypti mosquito larvae.

Keywords: blastospores, formulation, mosquitoes, conidia

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26 Controlling the Growth and Development of Mosquito (Aedes aegypti) Using Testosterone

Authors: Brian F. Estidola, Alfredo A. Alcantara, Catherine del Cruz, Genelita S. Garcia


This study aimed to investigate the effects of testosterone in the development and growth of Aedes aegypti as a main vector of dengue virus. There were three concentrations of testosterone: (0µM), (10µM), and (15µM) arranged randomly in two blocks. Each concentration houses 10 mosquitoes and monitored their development. The results showed that there were no significant differences on the effects of testosterone in emergence of larvae, mortality of eggs and larvae. However, it was shown that adults emerged from 15µM had a lower sex ratio than 10µM leading to the conclusion that there could be an optimal concentration of testosterone close to 10µM that could led to a high possibility of sex reversal of adult mosquitoes from female to male.

Keywords: mosquito, sex reversal, testosterone, ecdysterone

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25 An Ecofriendly Approach for the Management of Aedes aegypti L (Diptera: Culicidae) by Ocimum sanctum

Authors: Mohd Shazad, Kamal Kumar Gupta


Aedes aegypti (Diptera: Culicidae), commonly known as tiger mosquito is the vector of dengue fever, yellow fever, chikungunya and zika virus. In the absence of any effective vaccine against these diseases, control the mosquito population is the only promising mean to prevent the diseases. Currently used chemical insecticides cause environmental contamination, high mammalian toxicity and hazards to non-target organisms, insecticide resistance and vector resurgence. Present research work aimed to explore the potentials of phytochemicals present in the Ocimum sanctum in management of mosquito population. The leaves of Ocimum were extracted with ethanol by ‘cold extraction method’. 0-24h old fourth instar larvae of Aedes aegypti were treated with the extract of concentrations 50ppm, 100ppm, 200ppm and 400ppm for 24h. Survival, growth and development of the treated larvae were evaluated. The adults emerged from the treated larvae were used for the reproductive fitness studies. Our results indicate 77.2% mortality in the larvae exposed to 400 ppm. At lower doses, although there was no significant reduction in the survival after 24h however, it decreased during subsequent days of observations. In control experiments, no mortality was observed. It was also observed that the larvae survived after treatment showed severe growth and developmental abnormalities. There was significant increase in larval duration. In control, fourth instar moulted into pupa after 3 days while larvae treated with 400 ppm extract were moulted after 4.6 days. Larva-pupa intermediates and the pupa-adult intermediates were observed in many cases. The adults emerged from the treated larvae showed impaired mating and oviposition behaviour. The females exhibited longer preoviposition period, reduced oviposition rate and decreased egg output. GCMS analysis of the ethanol extract revealed presence of JH mimics and intermediates of JH biosynthetic pathway. Potentials of Ocimum sanctum in integrated vector management programme of Aedes aegypti were discussed.

Keywords: Aedes aegypti, Ocimum sanctum, oviposition, survival

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24 Effectiveness Assessment of a Brazilian Larvicide on Aedes Control

Authors: Josiane N. Muller, Allan K. R. Galardo, Tatiane A. Barbosa, Evan P. Ferro, Wellington M. Dos Santos, Ana Paula S. A. Correa, Edinaldo C. Rego, Jose B. P. Lima


The susceptibility status of an insect population to any larvicide depends on several factors such includes genetic constitution, environmental conditions and others. The mosquito Aedes aegypti is the primary vector of three important viral diseases, Zika, Dengue, and Chikungunya. The frequent outbreaks of those diseases in different parts of Brazil demonstrate the importance of testing the susceptibility of vectors in different environments. Since the control of this mosquito leads to the control of disease, alternatives for vector control that value the different Brazilian environmental conditions are needed for effective actions. The aim of this study was to evaluate a new commercial formulation of Bacillus thuringiensis israelenses (DengueTech: Brazilian innovative technology) in the Brazilian Legal Amazon considering the climate conditions. Semi-field tests were conducted in the Institute of Scientific and Technological Research of the State of Amapa in two different environments, one in a shaded area and the other exposed to sunlight. The mosquito larvae were exposed to larvicide concentration and a control; each group was tested in three containers of 40 liters each. To assess persistence 50 third instar larvae of Aedes aegypti laboratory lineages (Rockefeller) and 50 larvae of Aedes aegypti collected in the municipality of Macapa, Brazil’s Amapa state, were added weekly and after 24 hours the mortality was assessed. In total 16 tests were performed, where 12 were done with replacement of water (1/5 of the volume, three times per week). The effectiveness of the product was determined through mortality of ≥ 80%, as recommend by the World Health Organization. The results demonstrated that high-water temperatures (26-35 °C) on the containers influenced the residual time of the product, where the maximum effect achieved was 21 days in the shaded area; and no effectiveness of 60 days was found in any of the tests, as expected according to the larvicide company. The test with and without water replacement did not present significant differences in the mortality rate. Considering the different environments and climate, these results stimulate the need to test larvicide and its effectiveness in specific environmental settings in order to identify the parameters required for better results. Thus, we see the importance of semi-field researches considering the local climate conditions for a successful control of Aedes aegypti.

Keywords: Aedes aegypti, bioassay, larvicida, vector control

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23 Impact of the Achyranthes aspera (Amaranthaceae) Extracts on the Survival and Histological Architecture of the Midgut Epithelial Tissue of Early Fourth Instars of Aedes aegypti (Diptera: Culicidae)

Authors: Aarti Sharma, Sarita Kumar, Pushplata Tripathi


Aedes aegypti L. is one of the most important insect vectors in the world transmitting several diseases of concern; dengue fever, dengue haemorrhagic fever and yellow fever. Though since ages the control of dengue vector is primarily relied upon the use of synthetic chemical insecticides, the continued and indiscriminate use of insecticides for their control has received wide public apprehension because of multifarious problems including insecticide resistance, resurgence of pest species, environmental pollution, toxic hazards to humans and non-target organisms. These problems have necessitated the need to explore and develop alternative strategies using eco-friendly and bio-degradable plant products. Bio-insecticides, despite being the focus of research nowadays, have not been investigated much regarding their physiological effects on the mosquitoes. Thus, the present studies were carried out to investigate the anti-mosquito potential of the leaf and stem hexane extracts of Achyranthes aspera against early fourth instars of Aedes aegypti L and their effects on the histological architecture of their midgut. The larvicidal bioassays conducted with the A. aspera leaf hexane extracts revealed the respective LC30, LC50 and LC90 values of 66.545 ppm, 82.555 ppm, 139.817 ppm while the assays with stem hexane extracts resulted in respective values of 54.982 ppm, 68.133 ppm, 115.075 ppm. The studies clearly indicate the efficacy of extracts as larvicidal agents against Ae. aegypti, the stem extracts being found more effective than the leaf extracts. When the larvae assayed with extracts were investigated for the modifications in the histo-architecture of the midgut, the studies showed significant damage, shrinkage, distortion and vacuolization of gut tissues and peritrophic membrane causing disintegration of epithelial cells and cytoplasmic organelles; extent of toxicity and damage varied depending upon the concentration and exposure time period. These changes revealed appreciable stomach poison potential of A. aspera extracts against Ae. aegypti larvae, which may have also caused adverse impact on the growth and development of larvae. These effects were also found to be more pronounced with the stem extract than the leaf extract. Our findings may prove significant suggesting the use of A. aspera extract as a bio-insecticide against early fourth instar larvae of Ae. aegypti. Further studies are needed to identify the bioactive component in the extracts and to ascertain the use of component in the fields as anti-mosquito control agent.

Keywords: Achyranthes aspera, Aedes aegypti, histological architecture, larvicidal, midgut, stomach poison

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22 Studies on Knockdown Resistance Mutations in Aedes aegypti and Aedes albopictus in India

Authors: Neera Kapoor


Background: Knockdown Resistance (KDR) is one of the mechanisms of insecticide resistance in insects caused by the reduced target site sensitivity i.e. voltage gated sodium channel (VGSC) rendering it less sensitive to the toxic effects of DDT and pyrethroids. In this study, we evaluated insecticide susceptibility and its underlying KDR mechanism in eight Ae. aegypti and five Ae. albopictus field populations. Methodology: Field population was collected from four different geographical regions of India covering 18 districts of ten states. For genotyping of twelve KDR alleles in Ae. aegypti field populations, three PCR based assays were used; with DNA sequencing; ASPCR; PCR-RFLP. Genomic DNA was isolated, and three partial domains (II, III, and IV) of VGSC were amplified and sequenced. Results: Molecular screening for common KDR mutations, revealed the presence of five mutations viz. S989P, V1016G, T1520I, F1534C/L. Two novel mutations were observed, first at T1520 (ACC) residue where a C > T substitution at the second position of codon results in amino acid change to Isoleucine (ATC). Second mutation was an alternative point mutation at F1534 (TTC) residue where a substitution of T > C at the first position of codon results in an amino acid change to Leucine (CTC). ASPCRs were not accurate, so three PCR-RFLP assays were developed for genotyping of five KDR alleles in Ae. aegypti; viz. T1520I, F1534C/L. Representative samples of all genotypes (n=200) were sequenced to validate the newly developed PCR based assays for Ae. aegypti. Genotyping results showed that 989P is linked to 1016G and novel mutation 1520I was always found with 1534C allele. Conclusion: Present study confirmed the presence of DDT and pyrethroid resistance among Ae. aegypti populations in India and for the first time reported KDR mutations in this species from India including two novel mutations. Results of present study lead us to infer that, at least five KDR mutations (S989P, V1016G, T1530I, F1534C, and F1534L) can be seen as a potential marker for DDT/pyrethroid resistance.

Keywords: F1534C, F1534L, S989P, T1530I, V1016G

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21 The Effect of Extensive Mosquito Migration on Dengue Control as Revealed by Phylogeny of Dengue Vector Aedes aegypti

Authors: M. D. Nirmani, K. L. N. Perera, G. H. Galhena


Dengue has become one of the most important arbo-viral disease in all tropical and subtropical regions of the world. Aedes aegypti, is the principal vector of the virus, vary in both epidemiological and behavioral characteristics, which could be finely measured through DNA sequence comparison at their population level. Such knowledge in the population differences can assist in implementation of effective vector control strategies allowing to make estimates of the gene flow and adaptive genomic changes, which are important predictors of the spread of Wolbachia infection or insecticide resistance. As such, this study was undertaken to investigate the phylogenetic relationships of Ae. aegypti from Galle and Colombo, Sri Lanka, based on the ribosomal protein region which spans between two exons, in order to understand the geographical distribution of genetically distinct mosquito clades and its impact on mosquito control measures. A 320bp DNA region spanning from 681-930 bp, corresponding to the ribosomal protein, was sequenced in 62 Ae. aegypti larvae collected from Galle (N=30) and Colombo (N=32), Sri Lanka. The sequences were aligned using ClustalW and the haplotypes were determined with DnaSP 5.10. Phylogenetic relationships among haplotypes were constructed using the maximum likelihood method under Tamura 3 parameter model in MEGA 7.0.14 including three previously reported sequences of Australian (N=2) and Brazilian (N=1) Ae. aegypti. The bootstrap support was calculated using 1000 replicates and the tree was rooted using Aedes notoscriptus (GenBank accession No. KJ194101). Among all sequences, nineteen different haplotypes were found among which five haplotypes were shared between 80% of mosquitoes in the two populations. Seven haplotypes were unique to each of the population. Phylogenetic tree revealed two basal clades and a single derived clade. All observed haplotypes of the two Ae. aegypti populations were distributed in all the three clades, indicating a lack of genetic differentiation between populations. The Brazilian Ae. aegypti haplotype and one of the Australian haplotypes were grouped together with the Sri Lankan basal haplotype in the same basal clade, whereas the other Australian haplotype was found in the derived clade. Phylogram showed that Galle and Colombo Ae. aegypti populations are highly related to each other despite the large geographic distance (129 Km) indicating a substantial genetic similarity between them. This may have probably arisen from passive migration assisted by human travelling and trade through both land and water as the two areas are bordered by the sea. In addition, studied Sri Lankan mosquito populations were closely related to Australian and Brazilian samples. Probably this might have caused by shipping industry between the three countries as all of them are fully or partially enclosed by sea. For example, illegal fishing boats migrating to Australia by sea is perhaps a good mean of transportation of all life stages of mosquitoes from Sri Lanka. These findings indicate that extensive mosquito migrations occur between populations not only within the country, but also among other countries in the world which might be a main barrier to the successful vector control measures.

Keywords: Aedes aegypti, dengue control, extensive mosquito migration, haplotypes, phylogeny, ribosomal protein

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20 Analysing Time Series for a Forecasting Model to the Dynamics of Aedes Aegypti Population Size

Authors: Flavia Cordeiro, Fabio Silva, Alvaro Eiras, Jose Luiz Acebal


Aedes aegypti is present in the tropical and subtropical regions of the world and is a vector of several diseases such as dengue fever, yellow fever, chikungunya, zika etc. The growth in the number of arboviruses cases in the last decades became a matter of great concern worldwide. Meteorological factors like mean temperature and precipitation are known to influence the infestation by the species through effects on physiology and ecology, altering the fecundity, mortality, lifespan, dispersion behaviour and abundance of the vector. Models able to describe the dynamics of the vector population size should then take into account the meteorological variables. The relationship between meteorological factors and the population dynamics of Ae. aegypti adult females are studied to provide a good set of predictors to model the dynamics of the mosquito population size. The time-series data of capture of adult females of a public health surveillance program from the city of Lavras, MG, Brazil had its association with precipitation, humidity and temperature analysed through a set of statistical methods for time series analysis commonly adopted in Signal Processing, Information Theory and Neuroscience. Cross-correlation, multicollinearity test and whitened cross-correlation were applied to determine in which time lags would occur the influence of meteorological variables on the dynamics of the mosquito abundance. Among the findings, the studied case indicated strong collinearity between humidity and precipitation, and precipitation was selected to form a pair of descriptors together with temperature. In the techniques used, there were observed significant associations between infestation indicators and both temperature and precipitation in short, mid and long terms, evincing that those variables should be considered in entomological models and as public health indicators. A descriptive model used to test the results exhibits a strong correlation to data.

Keywords: Aedes aegypti, cross-correlation, multicollinearity, meteorological variables

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19 Assessing Antimicrobial Activity of Various Plant Extracts on Midgutmicroflora of Aedesaegypti

Authors: V. Baweja, K. K. Gupta, V. Dubey, C. Keshavam


Antimicrobial activity of six indigenous plants such as Tulsi Ocimum sanctum, Neem Azadirachta indica, Aloe vera, Turmeric Curcuma longa, Lantana Lantana camara, and Clove Syzygium aromaticum was assessed against the gut microbiota of the dengue fever mosquito Aedes aegypti, keeping in view that the presence of midgut bacteria may affect the ability of the vector to transmit pathogens. Eleven different types of bacterial clones were isolated from the midgut of lab-reared fourth instar larvae of Aedes aegypti and were grown on LB agar medium at an optimum temperature of 25 ºC. Identification of these bacteria was done on the basis of their colony characteristic such as colony size, shape, opacity, elevation, consistency, and growth. Light microscopic studies of the gut microbiota revealed dominance of Gram-negative cocci over gram positive cocci and bacilli and Gram-negative bacilli. Identification of species was done by chemical characterization of the colonies. Crude extracts of all test plants were screened for their antimicrobial activities against gut microbiota by disc diffusion assay. The zone of exclusion seen after 24 hr of incubation in different assays revealed the most potent antibacterial activities in neem followed by clove and turmeric. Lantana and Aloe vera were least effective.

Keywords: plant extract, aedes, dengue, antimicrobial activity

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18 A Versatile Algorithm to Propose Optimized Solutions to the Dengue Disease Problem

Authors: Fernando L. P. Santos, Luiz G. Lyra, Helenice O. Florentino, Daniela R. Cantane


Dengue is a febrile infectious disease caused by a virus of the family Flaviridae. It is transmitted by the bite of mosquitoes, usually of the genus Aedes aegypti. It occurs in tropical and subtropical areas of the world. This disease has been a major public health problem worldwide, especially in tropical countries such as Brazil, and its incidence has increased in recent years. Dengue is a subject of intense research. Efficient forms of mosquito control must be considered. In this work, the mono-objective optimal control problem was solved for analysing the dengue disease problem. Chemical and biological controls were considered in the mathematical aspect. This model describes the dynamics of mosquitoes in water and winged phases. We applied the genetic algorithms (GA) to obtain optimal strategies for the control of dengue. Numerical simulations have been performed to verify the versatility and the applicability of this algorithm. On the basis of the present results we may recommend the GA to solve optimal control problem with a large region of feasibility.

Keywords: genetic algorithm, dengue, Aedes aegypti, biological control, chemical control

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17 Pyrethroid and Organophosphate Susceptibility Status of Aedesaegypti (Linnaeus), Aedes albopictus (Skuse) and Culex quinquefasciatus (Say) in Penang, Malaysia

Authors: Hadura Abu Hasan, Zairi Jaal, P. J. McCall


Dengue is a serious problem in Malaysia, particularly in high-density urban communities with lower socio-economic levels. This study evaluated the susceptibility of local populations of Aedesaegypti (Linnaeus), Aedesalbopictus (Skuse) and Culexquinquefasciatus (Say) from the traditional community of BaganDalam, Penang, Malaysia to lambdacyhalothrin and pirimiphos-methyl using standard World Health Organization (WHO) adult bioassay test. Unfed female mosquitoes aged 3-5 days were exposed to WHO recommended dosages of insecticides over fixed time periods with results presented as knock-down time (KT50) for each strain.The insecticide susceptible VCRU laboratory strain was usedas control. All three specieswere highly resistant to lambda-cyhalothrin with less than 10% mortality at 24 hours after treatment. In contrast, Ae.aegypti and Ae. albopictus were susceptible to pirimiphos-methyl, showing 100% mortality recorded 24 hoursafter treatment. Cx. quinquefasciatuswasclassed as ‘suspected resistant’ to pirimiphos-methyl as mortality recorded 24 hours after treatment was 94-96%. The results indicate that organophosphates such as pirimiphos-methyl might be used as alternative to pyrethroid for dengue vector control in this dengue-prone area.

Keywords: vector control, aedes aegypti, aedes albopictus, dengue, culex quinquefasciatus, residuals insecticides, pyrethroid, organophosphate, resistant, mosquito

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16 DNA Barcoding for Identification of Dengue Vectors from Assam and Arunachal Pradesh: North-Eastern States in India

Authors: Monika Soni, Shovonlal Bhowmick, Chandra Bhattacharya, Jitendra Sharma, Prafulla Dutta, Jagadish Mahanta


Aedes aegypti and Aedes albopictus are considered as two major vectors to transmit dengue virus. In North-east India, two states viz. Assam and Arunachal Pradesh are known to be high endemic zone for dengue and Chikungunya viral infection. The taxonomical classification of medically important vectors are important for mapping of actual evolutionary trends and epidemiological studies. However, misidentification of mosquito species in field-collected mosquito specimens could have a negative impact which may affect vector-borne disease control policy. DNA barcoding is a prominent method to record available species, differentiate from new addition and change of population structure. In this study, a combined approach of a morphological and molecular technique of DNA barcoding was adopted to explore sequence variation in mitochondrial cytochrome c oxidase subunit I (COI) gene within dengue vectors. The study has revealed the map distribution of the dengue vector from two states i.e. Assam and Arunachal Pradesh, India. Approximate five hundred mosquito specimens were collected from different parts of two states, and their morphological features were compared with the taxonomic keys. The analysis of detailed taxonomic study revealed identification of two species Aedes aegypti and Aedes albopictus. The species aegypti comprised of 66.6% of the specimen and represented as dominant dengue vector species. The sequences obtained through standard DNA barcoding protocol were compared with public databases, viz. GenBank and BOLD. The sequences of all Aedes albopictus have shown 100% similarity whereas sequence of Aedes aegypti has shown 99.77 - 100% similarity of COI gene with that of different geographically located same species based on BOLD database search. From dengue prevalent different geographical regions fifty-nine sequences were retrieved from NCBI and BOLD databases of the same and related taxa to determine the evolutionary distance model based on the phylogenetic analysis. Neighbor-Joining (NJ) and Maximum Likelihood (ML) phylogenetic tree was constructed in MEGA6.06 software with 1000 bootstrap replicates using Kimura-2-Parameter model. Data were analyzed for sequence divergence and found that intraspecific divergence ranged from 0.0 to 2.0% and interspecific divergence ranged from 11.0 to 12.0%. The transitional and transversional substitutions were tested individually. The sequences were deposited in NCBI: GenBank database. This observation claimed the first DNA barcoding analysis of Aedes mosquitoes from North-eastern states in India and also confirmed the range expansion of two important mosquito species. Overall, this study insight into the molecular ecology of the dengue vectors from North-eastern India which will enhance the understanding to improve the existing entomological surveillance and vector incrimination program.

Keywords: COI, dengue vectors, DNA barcoding, molecular identification, North-east India, phylogenetics

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15 Screening for Larvicidal Activity of Aqueous and Ethanolic Extracts of Fourteen Selected Plants and Formulation of a Larvicide against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) Larvae

Authors: Michael Russelle S. Alvarez, Noel S. Quiming, Francisco M. Heralde


This study aims to: a) obtain ethanolic (95% EtOH) and aqueous extracts of Selaginella elmeri, Christella dentata, Elatostema sinnatum, Curculigo capitulata, Euphorbia hirta, Murraya koenigii, Alpinia speciosa, Cymbopogon citratus, Eucalyptus globulus, Jatropha curcas, Psidium guajava, Gliricidia sepium, Ixora coccinea and Capsicum frutescens and screen them for larvicidal activities against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) larvae; b) to fractionate the most active extract and determine the most active fraction; c) to determine the larvicidal properties of the most active extract and fraction against by computing their percentage mortality, LC50, and LC90 after 24 and 48 hours of exposure; and d) to determine the nature of the components of the active extracts and fractions using phytochemical screening. Ethanolic (95% EtOH) and aqueous extracts of the selected plants will be screened for potential larvicidal activity against Ae. aegypti and Ae. albopictus using standard procedures and 1% malathion and a Piper nigrum based ovicide-larvicide by the Department of Science and Technology as positive controls. The results were analyzed using One-Way ANOVA with Tukey’s and Dunnett’s test. The most active extract will be subjected to partial fractionation using normal-phase column chromatography, and the fractions subsequently screened to determine the most active fraction. The most active extract and fraction were subjected to dose-response assay and probit analysis to determine the LC50 and LC90 after 24 and 48 hours of exposure. The active extracts and fractions will be screened for phytochemical content. The ethanolic extracts of C. citratus, E. hirta, I. coccinea, G. sepium, M. koenigii, E globulus, J. curcas and C. frutescens exhibited significant larvicidal activity, with C. frutescens being the most active. After fractionation, the ethyl acetate fraction was found to be the most active. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, indoles and steroids. A formulation using talcum powder–300 mg fraction per 1 g talcum powder–was made and again tested for larvicidal activity. At 2 g/L, the formulation proved effective in killing all of the test larvae after 24 hours.

Keywords: larvicidal activity screening, partial purification, dose-response assay, capsicum frutescens

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14 The Effect of Aromatherapy Candle as Insecticide from Citrus Extract of Lemongrass (Cymbopogon) to Increase Ae. aegypti Mortality

Authors: Nurul Hidayah, Farida Rahmatika, Fathimah Azzahra, Nesty Herennadia


Aromatherapy candles are one of the insecticide media that have not been much researched. The active ingredient that is proven to have the effect of insecticide is a citrus extract from lemongrass oil (Cymbopogon). Aromatherapy candles are added by citrus compounds to be insecticidal for Ae. aegypti mosquito that was related to the infectious disease such as dengue fever. This research aims to find out if aromatherapy candles of citrus compounds have an insecticidal effect on Ae. aegypti mosquito. We used true experimental design including posttest only with control group design. The samples are 20 male and female Ae. aegypti mosquitos with aged 1-7 days belong to the inclusion criteria. The subjects were divided into 6 groups, consisting of 1 negative control group and 5 treatment groups with variation concentration are 1%; 2%; 3%; 4%; 5%. Each group will be treated for 2 hours and observed death after 24 hours. Replication in each group is done 4 times. The results were then tested statistically using Kruskal-Wallis and probit test. Mean of death in negative control group, and treatment group 1%; 2%; 3%; 4%; 5% respectively 0; 1; 0.25; 0; 1 and 1 mosquito. The Kruskal-Wallis test in the study group found no significant difference (p = 0.178). The probit analysis showed that LC50 and LC90 were 20.069% and 31.557%. The aromatherapy candle of a citrus compound has an insecticidal effect on the Ae aegypti mosquito.

Keywords: Ae. aegypti insecticide, aromatherapy candle, citrus compound, lemongrass oil (Cymbopogon)

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13 Study on the Incidence of Chikungunya Infection in Swat Region

Authors: Nasib Zaman, Maneesha Kour, Muhammad Rizwan, Fazal Akbar


Abstract: Chikungunya fever is a re-emerging rapidly spreading mosquito-borne disease cause by Aedes albopictus and Aedes aegypti mosquito vectors. Currently, it is affecting millions of people globally. Objective: This study's main objective was to find the incidence of chikungunya fever in the Swat region and the factors associated with the spread of this infection. Method: This study was carried out in different areas of Swat. Blood samples and data were collected from selected patients, and a questionnaire was filled for each patient. 3-5ml of the specimen was taken from the patient's vein and serum, or plasma was separated by centrifugation. Chikungunya tests were performed for IgG and IgM antibodies. The data was analyzed by SPSS and Graph Paid Prism 5. Results: A total of 169 patients were included in this study, out of which 103 (60.9%) having age less than 30 years were positive for chikungunya infection and 66 (39.1%) having more than 30 years were negative for this infection. Only 1 (0.6%) were positive for both IgG and IgM antibody. About 15 (8.9%) patients have diagnosed with positive IgG antibodies, and 25 (26.6%) patients were positive for IgM positive antibodies. The infection rate was significantly higher in males compared to females 71 (59.6%) vs. 14 (38%) P value=0.088, OR=1.7. Conclusion: This study concludes clinical knowledge and awareness that are necessary for a diagnosis of chikungunya infection properly. Therefore it is important to educate people for the eradication of this infection. Recommendation: This study also recommends investigating the other risk factors associated with this infection.

Keywords: Chikungunya, risk factor, Incidence, antibodies, mosquito

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12 Bioefficiency of Cinnamomum verum Loaded Niosomes and Its Microbicidal and Mosquito Larvicidal Activity against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus

Authors: Aasaithambi Kalaiselvi, Michael Gabriel Paulraj, Ekambaram Nakkeeran


Emergences of mosquito vector-borne diseases are considered as a perpetual problem globally in tropical countries. The outbreak of several diseases such as chikungunya, zika virus infection and dengue fever has created a massive threat towards the living population. Frequent usage of synthetic insecticides like Dichloro Diphenyl Trichloroethane (DDT) eventually had its adverse harmful effects on humans as well as the environment. Since there are no perennial vaccines, prevention, treatment or drugs available for these pathogenic vectors, WHO is more concerned in eradicating their breeding sites effectively without any side effects on humans and environment by approaching plant-derived natural eco-friendly bio-insecticides. The aim of this study is to investigate the larvicidal potency of Cinnamomum verum essential oil (CEO) loaded niosomes. Cholesterol and surfactant variants of Span 20, 60 and 80 were used in synthesizing CEO loaded niosomes using Transmembrane pH gradient method. The synthesized CEO loaded niosomes were characterized by Zeta potential, particle size, Fourier Transform Infrared Spectroscopy (FT-IR), GC-MS and SEM analysis to evaluate charge, size, functional properties, the composition of secondary metabolites and morphology. The Z-average size of the formed niosomes was 1870.84 nm and had good stability with zeta potential -85.3 meV. The entrapment efficiency of the CEO loaded niosomes was determined by UV-Visible Spectrophotometry. The bio-potency of CEO loaded niosomes was treated and assessed against gram-positive (Bacillus subtilis) and gram-negative (Escherichia coli) bacteria and fungi (Aspergillus fumigatus and Candida albicans) at various concentrations. The larvicidal activity was evaluated against II to IV instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus at various concentrations for 24 h. The mortality rate of LC₅₀ and LC₉₀ values were calculated. The results exhibited that CEO loaded niosomes have greater efficiency against mosquito larvicidal activity. The results suggest that niosomes could be used in various applications of biotechnology and drug delivery systems with greater stability by altering the drug of interest.

Keywords: Cinnamomum verum, niosomes, entrapment efficiency, bactericidal and fungicidal, mosquito larvicidal activity

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11 Insecticide Resistance Detection on Dengue Vector, Aedes albopictus Obtained from Kapit, Kuching and Sibu Districts in Sarawak State, Malaysia

Authors: Koon Weng Lau, Chee Dhang Chen, Abdul Aziz Azidah, Mohd Sofian-Azirun


Recently, Sarawak state of Malaysia encounter an outbreak of dengue fever. Aedes albopictus has incriminated as one of the important vectors of dengue transmission. Without an effective vaccine, approaches to control or prevent dengue will be a focus on the vectors. The control of Aedes mosquitoes is still dependent on the use of chemical insecticides and insecticide resistance represents a threat to the effectiveness of vector control. This study was conducted to determine the resistance status of 11 active ingredients representing four major insecticide classes: DDT, dieldrin, malathion, fenitrothion, bendiocarb, propoxur, etofenprox, deltamethrin, lambda-cyhalothrin, cyfluthrin, and permethrin. Standard WHO test procedures were conducted to determine the insecticide susceptibility. Aedes albopictus collected from Kapit (resistance ratio, RR = 1.04–3.02), Kuching (RR = 1.17–4.61), and Sibu (RR = 1.06–3.59) exhibited low resistance toward all insecticides except dieldrin. This study reveled that dieldrin is still effective against Ae. albopictus, followed by fenitrothion, cyfluthrin, and deltamethrin. In conclusion, Ae. albopictus in Sarawak exhibited different resistance levels toward various insecticides and alternative solutions should be implemented to prevent further deterioration of the condition.

Keywords: Aedes albopictus, dengue, insecticide resistance, Malaysia

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10 Women's Perceptions of Zika Virus Prevention Recommendations: A Tale of Two Cities within Fortaleza, Brazil

Authors: Jeni Stolow, Lina Moses, Carl Kendall


Zika virus (ZIKV) reemerged as a global threat in 2015 with Brazil at its epicenter. Brazilians have a long history of combatting Aedes aegypti mosquitos as it is a common vector for dengue, chikungunya, and yellow fever. As a response to the epidemic, public health authorities promoted ZIKV prevention behaviors such as mosquito bite prevention, reproductive counseling for women who are pregnant or contemplating pregnancy, pregnancy avoidance, and condom use. Most prevention efforts from Brazil focused on the mosquito vector- utilizing recycled dengue approaches without acknowledging the context in which women were able to adhere to these prevention messages. This study used qualitative methods to explore how women in Fortaleza, Brazil perceive ZIKV, the Brazilian authorities’ ZIKV prevention recommendations, and the feasibility of adhering to these recommendations. A core study aim was to look at how women perceive their physical, social, and natural environment as it impacts women’s ability to adhere to ZIKV prevention behaviors. A Rapid Anthropological Assessment (RAA) containing observations, informational interviews, and semi-structured in-depth interviews were utilized for data collection. The study utilized Grounded Theory as the systematic inductive method of analyzing the data collected. Interviews were conducted with 35 women of reproductive age (15-39 years old), who primarily utilize the public health system. It was found that women’s self-identified economic class was associated with how strongly women felt they could prevent ZIKV. All women interviewed technically belong to the C-class, the middle economic class. Although all members of the same economic class, there was a divide amongst participants as to who perceived themselves as higher C-class versus lower C-class. How women saw their economic status was dictated by how they perceived their physical, social, and natural environment. Women further associated their environment and their economic class to their likelihood of contracting ZIKV, their options for preventing ZIKV, their ability to prevent ZIKV, and their willingness to attempt to prevent ZIKV. Women’s perceived economic status was found to relate to their structural environment (housing quality, sewage, and locations to supplies), social environment (family and peer norms), and natural environment (wetland areas, natural mosquito breeding sites, and cyclical nature of vectors). Findings from this study suggest that women’s perceived environment and economic status impact their perceived feasibility and desire to attempt behaviors to prevent ZIKV. Although ZIKV has depleted from epidemic to endemic status, it is suggested that the virus will return as cyclical outbreaks like that seen with similar arboviruses such as dengue and chikungunya. As the next ZIKV epidemic approaches it is essential to understand how women perceive themselves, their abilities, and their environments to best aid the prevention of ZIKV.

Keywords: Aedes aegypti, environment, prevention, qualitative, zika

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9 Comparative Vector Susceptibility for Dengue Virus and Their Co-Infection in A. aegypti and A. albopictus

Authors: Monika Soni, Chandra Bhattacharya, Siraj Ahmed Ahmed, Prafulla Dutta


Dengue is now a globally important arboviral disease. Extensive vector surveillance has already established A.aegypti as a primary vector, but A.albopictus is now accelerating the situation through gradual adaptation to human surroundings. Global destabilization and gradual climatic shift with rising in temperature have significantly expanded the geographic range of these species These versatile vectors also host Chikungunya, Zika, and yellow fever virus. Biggest challenge faced by endemic countries now is upsurge in co-infection reported with multiple serotypes and virus co-circulation. To foster vector control interventions and mitigate disease burden, there is surge for knowledge on vector susceptibility and viral tolerance in response to multiple infections. To address our understanding on transmission dynamics and reproductive fitness, both the vectors were exposed to single and dual combinations of all four dengue serotypes by artificial feeding and followed up to third generation. Artificial feeding observed significant difference in feeding rate for both the species where A.albopictus was poor artificial feeder (35-50%) compared to A.aegypti (95-97%) Robust sequential screening of viral antigen in mosquitoes was followed by Dengue NS1 ELISA, RT-PCR and Quantitative PCR. To observe viral dissemination in different mosquito tissues Indirect immunofluorescence assay was performed. Result showed that both the vectors were infected initially with all dengue(1-4)serotypes and its co-infection (D1 and D2, D1 and D3, D1 and D4, D2 and D4) combinations. In case of DENV-2 there was significant difference in the peak titer observed at 16th day post infection. But when exposed to dual infections A.aegypti supported all combinations of virus where A.albopictus only continued single infections in successive days. There was a significant negative effect on the fecundity and fertility of both the vectors compared to control (PANOVA < 0.001). In case of dengue 2 infected mosquito, fecundity in parent generation was significantly higher (PBonferroni < 0.001) for A.albopicus compare to A.aegypti but there was a complete loss of fecundity from second to third generation for A.albopictus. It was observed that A.aegypti becomes infected with multiple serotypes frequently even at low viral titres compared to A.albopictus. Possible reason for this could be the presence of wolbachia infection in A.albopictus or mosquito innate immune response, small RNA interference etc. Based on the observations it could be anticipated that transovarial transmission may not be an important phenomenon for clinical disease outcome, due to the absence of viral positivity by third generation. Also, Dengue NS1 ELISA can be used for preliminary viral detection in mosquitoes as more than 90% of the samples were found positive compared to RT-PCR and viral load estimation.

Keywords: co-infection, dengue, reproductive fitness, viral quantification

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8 Dengue Prevention and Control in Kaohsiung City

Authors: Chiu-Wen Chang, I-Yun Chang, Wei-Ting Chen, Hui-Ping Ho, Ruei-Hun Chang, Joh-Jong Huang


Kaohsiung City is located in the tropical region where has Aedes aegypti and Aedes albopictus distributed; once the virus invades, it’s can easily trigger local epidemic. Besides, Kaohsiung City has a world-class airport and harbor, trade and tourism are close and frequently with every country, especially with the Southeast Asian countries which also suffer from dengue. Therefore, Kaohsiung City faces the difficult challenge of dengue every year. The objectives of this study was to enhance dengue clinical care, border management and vector surveillance in Kaohsiung City by establishing an larger scale, innovatively and more coordinated dengue prevention and control strategies in 2016, including (1) Integrated medical programs: facilitated 657 contract medical institutions, widely set up NS1 rapid test in clinics, enhanced triage and referrals system, dengue case daily-monitoring management (2) Border quarantine: comprehensive NS1 screening for foreign workers and fisheries when immigration, hospitalization and isolation for suspected cases, health education for high risk groups (foreign students, other tourists) (3) Mosquito control: Widely use Gravitrap to monitor mosquito density in environment, use NS1 rapid screening test to detect community dengue virus (4) Health education: create a dengue app for people to immediately inquire the risk map and nearby medical resources, routine health education to all districts to strengthen public’s dengue knowledge, neighborhood cleaning awards program. The results showed that after new integration of dengue prevention and control strategies fully implemented in Kaohsiung City, the number of confirmed cases in 2016 declined to 342 cases, the majority of these cases are the continuation epidemic in 2015; in fact, only two cases confirmed after the 2016 summer. Besides, the dengue mortality rate successfully decreased to 0% in 2016. Moreover, according to the reporting rate from medical institutions in 2014 and 2016, it dropped from 27.07% to 19.45% from medical center, and it decreased from 36.55% to 29.79% from regional hospital; however, the reporting rate of district hospital increased from 11.88% to 15.87% and also increased from 24.51% to 34.89% in general practice clinics. Obviously, it showed that under the action of strengthening medical management, it reduced the medical center’s notification ratio and improved the notification ratio of general clinics which achieved the great effect of dengue clinical management and dengue control.

Keywords: dengue control, integrated control strategies, clinical management, NS1

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7 A Review on Biological Control of Mosquito Vectors

Authors: Asim Abbasi, Muhammad Sufyan, Iqra, Hafiza Javaria Ashraf


The share of vector-borne diseases (VBDs) in the global burden of infectious diseases is almost 17%. The advent of new drugs and latest research in medical science helped mankind to compete with these lethal diseases but still diseases transmitted by different mosquito species, including filariasis, malaria, viral encephalitis and dengue are serious threats for people living in disease endemic areas. Injudicious and repeated use of pesticides posed selection pressure on mosquitoes leading to development of resistance. Hence biological control agents are under serious consideration of scientific community to be used in vector control programmes. Fish have a history of predating immature stages of different aquatic insects including mosquitoes. The noteworthy examples in Africa and Asia includes, Aphanius discolour and a fish in the Panchax group. Moreover, common mosquito fish, Gambusia affinis predates mostly on temporary water mosquitoes like anopheline as compared to permanent water breeders like culicines. Mosquitoes belonging to genus Toxorhynchites have a worldwide distribution and are mostly associated with the predation of other mosquito larvae habituating with them in natural and artificial water containers. These species are harmless to humans as their adults do not suck human blood but feeds on floral nectar. However, their activity is mostly temperature dependent as Toxorhynchites brevipalpis consume 359 Aedes aegypti larvae at 30-32 ºC in contrast to 154 larvae at 20-26 ºC. Although many bacterial species were isolated from mosquito cadavers but those belonging to genus Bacillus are found highly pathogenic against them. The successful species of this genus include Bacillus thuringiensis and Bacillus sphaericus. The prime targets of B. thuringiensis are mostly the immatures of genus Aedes, Culex, Anopheles and Psorophora while B. sphaericus is specifically toxic against species of Culex, Psorophora and Culiseta. The entomopathogenic nematodes belonging to family, mermithidae are also pathogenic to different mosquito species. Eighty different species of mosquitoes including Anopheles, Aedes and Culex proved to be highly vulnerable to the attack of two mermithid species, Romanomermis culicivorax and R. iyengari. Cytoplasmic polyhedrosis virus was the first described pathogenic virus, isolated from the cadavers of mosquito specie, Culex tarsalis. Other viruses which are pathogenic to culicine includes, iridoviruses, cytopolyhedrosis viruses, entomopoxviruses and parvoviruses. Protozoa species belonging to division microsporidia are the common pathogenic protozoans in mosquito populations which kill their host by the chronic effects of parasitism. Moreover, due to their wide prevalence in anopheline mosquitoes and transversal and horizontal transmission from infected to healthy host, microsporidia of the genera Nosema and Amblyospora have received much attention in various mosquito control programmes. Fungal based mycopesticides are used in biological control of insect pests with 47 species reported virulent against different stages of mosquitoes. These include both aquatic fungi i.e. species of Coelomomyces, Lagenidium giganteum and Culicinomyces clavosporus, and the terrestrial fungi Metarhizium anisopliae and Beauveria bassiana. Hence, it was concluded that the integrated use of all these biological control agents can be a healthy contribution in mosquito control programmes and become a dire need of the time to avoid repeated use of pesticides.

Keywords: entomopathogenic nematodes, protozoa, Toxorhynchites, vector-borne

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6 A Comparative Study of Dengue Fever in Taiwan and Singapore Based on Open Data

Authors: Wei Wen Yang, Emily Chia Yu Su


Dengue fever is a mosquito-borne tropical infectious disease caused by the dengue virus. After infection, symptoms usually start from three to fourteen days. Dengue virus may cause a high fever and at least two of the following symptoms, severe headache, severe eye pain, joint pains, muscle or bone pain, vomiting, feature skin rash, and mild bleeding manifestation. In addition, recovery will take at least two to seven days. Dengue fever has rapidly spread in tropical and subtropical areas in recent years. Several phenomena around the world such as global warming, urbanization, and international travel are the main reasons in boosting the spread of dengue. In Taiwan, epidemics occur annually, especially during summer and fall seasons. On the other side, Singapore government also has announced the amounts number of dengue cases spreading in Singapore. As the serious epidemic of dengue fever outbreaks in Taiwan and Singapore, countries around the Asia-Pacific region are becoming high risks of susceptible to the outbreaks and local hub of spreading the virus. To improve public safety and public health issues, firstly, we are going to use Microsoft Excel and SAS EG to do data preprocessing. Secondly, using support vector machines and decision trees builds predict model, and analyzes the infectious cases between Taiwan and Singapore. By comparing different factors causing vector mosquito from model classification and regression, we can find similar spreading patterns where the disease occurred most frequently. The result can provide sufficient information to predict the future dengue infection outbreaks and control the diffusion of dengue fever among countries.

Keywords: dengue fever, Taiwan, Singapore, Aedes aegypti

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5 An Assessment of Health Hazards in Urban Communities: A Study of Spatial-Temporal Variations of Dengue Epidemic in Colombo, Sri Lanka

Authors: U. Thisara G. Perera, C. M. Kanchana N. K. Chandrasekara


Dengue is an epidemic which is spread by Aedes Egyptai and Aedes Albopictus mosquitoes. The cases of dengue show a dramatic growth rate of the epidemic in urban and semi urban areas spatially in tropical and sub-tropical regions of the world. Incidence of dengue has become a prominent reason for hospitalization and deaths in Asian countries, including Sri Lanka. During the last decade the dengue epidemic began to spread from urban to semi-urban and then to rural settings of the country. The highest number of dengue infected patients was recorded in Sri Lanka in the year 2016 and the highest number of patients was identified in Colombo district. Together with the commercial, industrial, and other supporting services, the district suffers from rapid urbanization and high population density. Thus, drainage and waste disposal patterns of the people in this area exert an additional pressure to the environment. The district is situated in the wet zone and thus low lying lands constitute the largest portion of the district. This situation additionally facilitates mosquito breeding sites. Therefore, the purpose of the present study was to assess the spatial and temporal distribution patterns of dengue epidemic in Kolonnawa MOH area (Medical Officer of Health) in the district of Colombo. The study was carried out using 615 recorded dengue cases in Kollonnawa MOH area during the south east monsoon season from May to September 2016. The Moran’s I and Kernel density estimation were used as analytical methods. The analysis of data was accomplished through the integrated use of ArcGIS 10.1 software packages along with Microsoft Excel analytical tool. Field observation was also carried out for verification purposes during the study period. Results of the Moran’s I index indicates that the spatial distribution of dengue cases showed a cluster distribution pattern across the area. Kernel density estimation emphasis that dengue cases are high where the population has gathered, especially in areas comprising housing schemes. Results of the Kernel Density estimation further discloses that hot spots of dengue epidemic are located in the western half of the Kolonnawa MOH area, which is close to the Colombo municipal boundary and there is a significant relationship with high population density and unplanned urban land use practices. Results of the field observation confirm that the drainage systems in these areas function poorly and careless waste disposal methods of the people further encourage mosquito breeding sites. This situation has evolved harmfully from a public health issue to a social problem, which ultimately impacts on the economy and social lives of the country.

Keywords: Dengue epidemic, health hazards, Kernel density, Moran’s I, Sri Lanka

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4 Current Status of Mosquitoes Vector Research and Control in Iran

Authors: Seyed Hassan Moosa-kazemi, Hassan Vatandoost


Malaria, Dirofilaria immitis (dog heart worm), and D. repens (dirofilariasis), which are transmitted by mosquitoes, have been reported in Iran. The Iranian mosquito fauna includes seven genera, 65 species, and three subspecies. Aedes albopictus has been reported since. West Nile, Sindbis, Dengue, Japanese encephalitis viruses, and the nematode Setaria (setariasis) has been reported in the country but there are no information about their vectors in Iran. Iran is malaria elimination phase. Insecticides residual spraying (IRS), distributed of insecticides long lasting treated nets (ITNs), fogging, release of larvivours fishes and Bacillus thuringiensis, chemical larviciding, as well as case finding and manipulation and modification of breeding places carried out thought the IVM program in the country. Prolonged exposure to insecticides over several generations of the vectors, develop resistance, a capacity to survive contact with insecticides. However, use of insecticides in agriculture has often been implicated as contributing to resistance in mosquito’s vectors. Resistance of mosquitoes to some insecticides has been documented just within a few years after the insecticides were introduced. Some enzymes such as monooxygenases, esterases and glutathione S-transferases have been considered as a reason for resistance to pyrethroid insecticides. In conclusion, regarding to documented resistance and tolerance of mosquitoes vectors to some insecticides, resistance management is suggested by using new insecticide with novel mode of action.

Keywords: control, Iran, resistance, vector

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3 Entomological Survey of Mosquitoes Responsible for the Transmission of Lymphatic Filariasis in Biase Cross River State, Nigeria

Authors: Maurice Mbah


Entomological survey of mosquitoes responsible for the transmission of lymphatic filariasis in Biase Local Government area of Cross River State, Nigeria within March and June 2017. Lymphatic filariasis is a mosquito-borne parasitic disease that is caused by three species of tissue dwelling filaroids (Wuchereria bancrofti; Brugia malayi; Brugia timori): Wuchereria bancrofti is responsible for 90% of cases and is found throughout the tropics and in some sub-tropical areas worldwide. The mosquitoes were caught using human landing catches, and pyrethrum spray catches method. The entomological analysis of mosquitoes which include speciating into genus and dissecting them to unveil any microfilaria in the thoracic region, abdomen, and mouth parts of the mosquitoes. Entomological analysis shows that, from the 1296 mosquitoes caught 795 (61.3%) were Culex species, 342 (26.4%) Anopheles species, 102 (7.9%) Aedes species, and 57 (4.4%) of other Genera. There was a statistically significant difference in the number of mosquitoes caught in the dry and rainy season (X²=0.62, P < 0.05). Out of 1213 mosquitoes dissected, 24(0.02%) contained developed stages (L₁ – L₃) of W. bancrofti larvae. 13 (0.01%) of the infected mosquitoes were of Culex species, and Anopheles species accounted for the other 11 (0.009%). There was a statistically significant difference in the infection rate between the two seasons (X²=0.87, P < 0.05). The correlation analysis showed a positive correlation between the infection rate among mosquitoes in the dry and rainy season (r=0.85, P < 0.05). The entomological studies showed that Anopheles species and the Culex species are the vectors of lymphatic filariasis in the study area.

Keywords: entomological survey, mosquitoes, lymphatic filariasis, biase

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2 Using Baculovirus Expression Vector System to Express Envelop Proteins of Chikungunya Virus in Insect Cells and Mammalian Cells

Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu


Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

Procedia PDF Downloads 351