Screening for larvicidal activity in some Thai plants against four mosquito vector species

Ninety-six ethanolic extracts from various parts of 84 Thai plant species were tested for their larvicidal activity against Aedes aegypti mosquitoes. Extracts from Rhinacanthus nasutus, Derris elliptica, Trigonostemon reidioides, Homalomena aromatica, Stemona tuberosa and Acorus calamus possessed high larvicidal activity, with LC50 values between 16.0 and 48.2 mg/l. Petroleum ether (PE) and methanol (MeOH) extracts were tested for their larvicidal activity against 4 mosquito vector species. The PE extract of R. nasutus exhibited larvicidal effects against Ae. aegypti, Culex quinquefasciatus, Anopheles dirus and Mansonia uniformis with LC50 values between 3.9 and 11.5 mg/l, while the MeOH extract gave LC50 values of between 8.1 and 14.7 mg/l. D. elliptica PE extract showed LC50 values of between 11.2 and 18.84 mg/l and the MeOH extract exhibited LC50 values between 13.2 and 45.2 mg/l.     

Coverage-Dependent Effect of Insecticide-Treated Curtains for Dengue Control in Thailand

Evidence on the effectiveness of insecticide-treated curtains (ITCs) for reducing densities of Aedes mosquitoes, the principal vectors of dengue, is scarce. In Laem Chabang southeast of Bangkok, Thailand, the Breteau Index (BI) (number of positive containers/100 houses) was 45 in October 2006. In March 2007, we distributed long-lasting ITCs in 22 clusters (2,032 houses) and selected 66 control clusters (661 houses). Routine control activities continued in all clusters. Six months after distribution, the BI was 25.8 and 77.6 in intervention and control areas, respectively (P < 0.001). Eighteen months after distribution, the BI was 21.8 and 23.8, respectively (P = 0.28). The average number of ITCs/house at cluster level was associated with the BI (P < 0.01) after six months, when 70.5% of households still used ITCs, but not at 18 months, when ITC coverage had decreased to 33.2%. Deployment of ITCs can result in considerable reductions in Aedes infestation levels, but the effect is coverage dependent.     

Laboratory colonization of Mansonia mosquitoes with an emphasis on Ma. annulata and Ma. bonneae

The present study records the first successful colonization of Mansonia annulata and describes colony maintenance with modification of rearing medium and host plants. Three species of Mansonia mosquitoes (Ma. uniformis, Ma. indiana and Ma annulifera) were successfully reared in ambient environments with adult emergence rates > 50%, while Ma. bonneae and Ma. dives yielded emergence rates > 30%. Colonization of Ma. annulata was modified and improved so that they were successfully raised to adult with emergence rates of 23%. Tube sedge, Lepironia articulata, was utilized as a host plant and peat swamp water was used as a rearing medium. Yeast and small lizard droppings were added daily to the larval medium to maintain microorganisms and pH in the infusion. However, identifying suitable culture medium remains an obstacle to establishing colonies of Ma. annulata, as the culture medium is difficult to mimic in the laboratory. Further study, focusing particularly on larval attachment substrates and rearing medium, is needed to develop a standardized and practical rearing technique for Mansonia mosquitoes.     

Bionomics studies of Mansonia mosquitoes inhabiting the peat swamp forest

The present study was conducted in the years 2000-2002 to determine the bionomics of Mansonia mosquitoes, vectors of nocturnally subperiodic Brugia malayi, inhabiting the peat swamp forest, "Phru Toh Daeng", Narathiwat Province, Thailand. Fifty-four species of mosquitoes belonging to 12 genera were added, for the first time, to the list of animal fauna in the peat swamp forest. Mansonia mosquitoes were the most abundant (60-70%) by all collection methods and occurred throughout the year with a high biting density (10.5-57.8 bites per person-hour). Ma. bonneae was most prevalent (47.5%) and fed on a variety of animal hosts, including domestic cats, cows, monkeys, and man with a maximum biting density of 24.3 bites per person-hour in October. The infective bites were found for the first time in Ma. annulata collected at Ban Toh Daeng (13 00-14 00 hours) and also Ma. bonneae at forest shade (16 00-17 00 hours) and in a village (20 00-21 00 hours) with rates of 0.6, 1.1 and 1.0%, respectively. The biting activities of these two species occurred in both the day and night time, with two lower peaks at 10 00 hours (18.5 bites per person-hour) and 13 00-15 00 (8.5-10.0 bites per person-hour) hours, but the highest peak was 19 00-21 00 hours (31.5-33.0 bites per person-hour) The biting activity patterns corresponded with the periodicity found in man and domestic cats and may play an important role in either transmission or maintenance of the filarial parasites in the peat swamp forest. The relative role of Ma. bonneae and Ma. uniformis in different environmental settings (primary swamp forest and open swamp) on the transmission of nocturnally subperiodic B. malayi merits further study.     

Lead levels of Culex mosquito larvae inhabiting lead utilizing factory

Objective

To determine lead level primarily in Culex quinquefasciatus (Cx. quinquefasciatus), and Culex gelidus (Cx. gelidus) larvae inhabiting lead consuming factories, and to putatively estimate eco-toxicological impact of effluents from the firms.

Methods

Third instars larvae were sampled by standard dipping method and lead concentrations in the larvae and their respective surrounding factory aquatic environments were determined through standard atomic absorption spectrophotometry (AAS).

Results

Cx. quinquefasciatus was the most abundant species followed by Cx. gelidus. The levels of lead were higher in the Cx. quinquefasciatus (1.08-47.47 µg/g), than in the wastewaters surface (0.01-0.78 µg/mL) from the factories or closer areas around factories. Other species were not reaching the criteria for lead determination.

Conclusions

The Cx. quinquefasciatus larvae can bio-accumulate the metal and can potentially serve as a biomarker of lead contamination, to complemente conventional techniques.     

Cytogenetic and molecular evidence for an additional new species within the taxon Anopheles barbirostris (Diptera: Culicidae) in Thailand

ITS2 DNA sequences of 42 isoline colonies of Anopheles barbirostris species A1 and A2 were analyzed and a new genetic species, temporarily designated as species A4 (Chiang Mai), was revealed. The large sequence divergences of the ITS2 (0.116-0.615), COI (0.023–0.048), and COII (0.030–0.040) genes between A. barbirostris species A4/A1 (Chiang Mai), A4/A2 (Phetchaburi), A4/A3 (Kanchanaburi), and A4/Anopheles campestris-like Form E (Chiang Mai) provided good supporting evidence. Species A1, A2, A3, and A4 share a mitotic karyotype of Form A (X₁, X₂, Y₁). Crossing experiments between species A4 and the other four species yielded strong reproductive isolation producing few and/or non-hatched eggs and inviable and/or abnormal development of the reproductive system of F₁ progenies. Moreover, available F₁ hybrid larvae showed asynaptic polytene chromosome arms. Hence, molecular and cytogenetic evidence strongly support the existence of A. barbirostris species A4, which is more closely related to A. campestris-like Form E than to species A1, A2, and A3. Additionally, crossing experiments among 12 and seven isolines of different cytological forms of species A1 (A, B, C, D) and A2 (A, B), respectively, yielded fertile and viable F₁ progenies. Thus, different karyotypic forms occurring in natural populations of species A1 and A2 merely represent intraspecies variation of sex chromosomes due to the extra blocks of heterochromatin.     

Comparative repellency of 38 essential oils against mosquito bites

The mosquito repellent activity of 38 essential oils from plants at three concentrations was screened against the mosquito Aedes aegypti under laboratory conditions using human subjects. On a volunteer's forearm, 0.1 mL of oil was applied per 30 cm2 of exposed skin. When the tested oils were applied at a 10% or 50% concentration, none of them prevented mosquito bites for as long as 2 h, but the undiluted oils of Cymbopogon nardus (citronella), Pogostemon cablin (patchuli), Syzygium aromaticum (clove) and Zanthoxylum limonella (Thai name: makaen) were the most effective and provided 2 h of complete repellency.From these initial results, three concentrations (10%, 50% and undiluted) of citronella, patchouli, clove and makaen were selected for repellency tests against Culex quinquefasciatus and Anopheles dirus. As expected, the undiluted oil showed the highest protection in each case. Clove oil gave the longest duration of 100% repellency (2-4 h) against all three species of mosquito.     

Potential for Anopheles campestris (Diptera: Culicidae) to transmit malaria parasites in Pa Rai subdistrict (Aranyaprathet,Sa Kaeo Province), Thailand

Member(s) of the Anopheles barbirostris group Reid, particularly Anopheles barbirostris and Anopheles campestris Reid are the suspected vectors of Plasmodium vivax in Pa Rai (Aranyaprathet, Sa Kaeo province). To determine if An. barbirostris, An. campestris, or both, are present in Pa Rai and to determine their potential to transmit malaria, a field and laboratory study was conducted. Isofemale colonizations of wild caught mosquitoes captured by landing catches were made for species confirmation and to determine the mosquito life cycle. Pupal morphology indicated all mosquitoes were An. campestris. During the late rainy season (October and November), An. campestris populations comprised 78.6% of all females captured by human landing catches and 7.1% of mosquitoes in a cow-baited trap. The biting activity cycle peaked between 2000 and 0100 hours and was highest (17.6 bites per person per hour) at 2300 hours. More An. campestris bit people indoors (nine bites per person per hour) than outdoors (four bites per person per hour). Immature An. campestris were found in ponds, swamps, rice-fields, puddles, marshes, ground pools, and pits with open sunlight to partial shade. The time from egg hatch to adult was 18-47 d and 14-22 d under laboratory (25.0-27.0 degrees C) and ambient (26-32 degrees C) conditions, respectively. The fecundity of An. campestris ranged from 173 to 311 eggs. Based on experimental infections, An. campestris was able to support the sporogonic cycle of P. vivax with 76.2 and 23.8% oocyst and sporozoite formation rate, respectively. An. campestris shows high potential as a malaria vector in Pa Rai.     

Esterase patterns of Anopheles dirus (Bangkok strain) and Anopheles balabacensis (Perlis form) in the laboratory

Esterase isoenzyme were performed in Anopheles balabacensis (Perlis Form) and Anopheles dirus (Bangkok Strain and Kanchanaburi Strain). The zymogram showed four positions, E1, E2, E3 and E4. Seventy-five percent of An. balabacensis (Perlis Form) or An. dirus (Bangkok Strain) can be correctly diagnosed by the presence of fast band in E3 or super slow band in E3 respectively. Homozygous slow band of E3 cannot be fully distinguished in both species, but by the combination of E2 and E3, 99.42% can be correctly identified to differentiate species.     

Development and application of a simple colorimetric assay reveals widespread distribution of sodium channel mutations in Thai populations of Aedes aegypti

Dengue fever and its more serious complications dengue haemorrhagic fever and dengue shock syndrome are growing public health problems in tropical and subtropical countries. In the absence of a vaccine, most dengue control programmes rely heavily on the use of insecticides to target the Aedes mosquito vectors. As a limited number of insecticides are routinely used in control, monitoring for the presence of resistance is an essential component of dengue prevention programmes. The pyrethroid insecticides target the voltage-gated sodium channel on the insects’ neurons. Substitutions at residue 1016 of this protein have been associated with pyrethroid and DDT resistance in Aedes aegypti populations from Latin America and Asia. Here we report on the development of a simple colorimetric assay to detect these mutations in individual mosquitoes. Evaluation of this diagnostic assay on 180 Ae. aegypti individuals from Thailand revealed the presence of high frequencies of the Val1016Gly mutation throughout the country. The assay requires no specialised equipment and will enable monitoring for insecticide resistance associated alleles to be routinely incorporated into dengue surveillance operations.