Aedes aegypti pupal/demographic surveys in southern Mexico: consistency and practicality.

In interventions aimed at the control of the immature stages of Aedes aegypti (L.), the principal vector of the dengue viruses, attempts are often made to treat or manage all larval habitats in households. When there are resource-constraints, however, a concentration of effort on the types of container that produce the most pupae may be required. Identification of these 'key' container types requires surveys of the immature stages and particularly - since these give the best estimates of the numbers of adults produced - of the numbers of pupae in local containers. Although there has been no clearly defined or standardized protocol for the sampling of Ae. aegypti pupae for many years, a methodology for 'pupal/demographic' surveys, which may allow the risk of dengue outbreaks in a given setting to be estimated, has been recently described. The consistency and practicality of using such surveys has now been investigated in three cities in the Mexican state of Chiapas, Mexico. Using a combination of 'quadrat'- and transect-sampling methods, 600 houses in each city were each sampled twice. Containers within each study household were searched for pupae and larvae. Although 107,297 containers, belonging to 26 categories, were observed, only 16,032 were found to contain water and 96% and 92% of these 'wet' containers contained no pupae and no third- or fourth-instar larvae, respectively. Although the random 'quadrat' sampling gave similar results to sampling along transects, there were statistically significant differences in the numbers of pupae according to container type and locality. The most important containers for pupal production were found to be large cement wash basins, which were present in almost every household investigated and from which 84% (10,257/12,271) of all pupae were collected. A focus on this class of container could serve as the basis of a targeted intervention strategy. When traditional Stegomyia indices were calculated they appeared to be correlated with the assessments of pupal abundance. The methodology for pupal/demographic surveys appears to be practical and to give consistent results, although it remains to be seen if monitoring of pupal productivity can adequately reflect the impact of vector-control interventions.

Effect of Novaluron (Rimon 10 EC) on the mosquitoes Anopheles albimanus, Anopheles pseudopunctipennis, Aedes aegypti, Aedes albopictus and Culex quinquefasciatus from Chiapas, Mexico.

Dengue fever is a serious problem in Mexico and vector control has not been effective enough at preventing outbreaks. Malaria is largely under control, but it is important that new control measures continue to be developed. Novaluron, a novel host-specific insect growth regulator and chitin synthesis inhibitor, has proved to be effective against agricultural pests, but its efficacy against larval mosquito vectors under field conditions remains unknown. In accordance with the World Health Organization Pesticide Evaluation Scheme, phase I, II and III studies were conducted to evaluate the efficacy and residual effect of Novaluron (Rimon 10 EC, Makhteshim, Beer-Sheva, Israel) on the malaria vectors Anopheles albimanus Wiedemann (Diptera: Culicidae) and Anopheles pseudopunctipennis Theobald, the dengue vectors Aedes aegypti (L) and Aedes albopictus Skuse and the nuisance mosquito Culex quinquefasciatus Say. Laboratory susceptibility tests yielded diagnostic concentrations for all five target species. Field trials to identify the optimum field dosage of Novaluron against Anopheles mosquitoes were carried out under semi-natural conditions in artificial plots and in vessels with wild mosquitoes. Efficacy was measured by monitoring mortality of larvae and pupae and the percentage of inhibition of emergence from floating cages. Dosages of Novaluron for field tests were based on pupal LC(99) (lethal concentration 99%) of An. pseudopunctipennis (0.166 mg/L) in plots and average pupal LC(99) of Ae. aegypti and Ae. albopictus (0.55 mg/L). At all dosages tested, Novaluron significantly reduced larval populations of An. albimanus, Culex coronator Dyar & Knab, Ae. albopictus and Cx. quinquefasciatus by approximately 90%, inhibited adult emergence of An. albimanus and An. pseudopunctipennis by approximately 97% for almost 4 months in experimental plots, and inhibited adult emergence of Ae. aegypti and Ae. albopictus by approximately 97% for up to 14 weeks. Recommended dosages of Novaluron for non-container breeding and container breeding mosquitoes are 0.166 mg/L and 0.55 mg/L, respectively. Overall, the residual effect was more sustained than that of temephos. The lowest dosage of Novaluron had less of an impact on non-target organisms than did temephos. Small-scale field trials in natural breeding sites treated with Novaluron at 0.6 L/ha eliminated adult emergence of An. albimanus and Cx. coronator for 8 weeks. For phase III studies, Novaluron was tested at the local and village levels, applying the optimum field rate to all natural breeding habitats within 1 km of a pair of neighbouring villages. Village-scale trials of Novaluron at 0.6 L/ha reduced An. albimanus larval populations for at least 8 weeks and, more importantly, sharply reduced the densities of adult host-seeking mosquitoes approaching houses. We conclude that Novaluron is effective and environmentally safer than temephos.

Sublethal iridovirus disease of the mosquito Aedes aegypti is due to viral replication not cytotoxicity.

Invertebrate iridescent viruses (Iridoviridae) possess a highly cytotoxic protein. In mosquitoes (Diptera: Culicidae), invertebrate iridescent virus 6 (IIV-6) usually causes covert (inapparent) infection that reduces fitness. To determine whether sublethal effects of IIV-6 are principally due to cytotoxicity of the viral inoculum (which inhibits macromolecular synthesis in the host), or caused by replication of the virus larvae of the mosquito Aedes aegypti (L) were exposed to untreated IIV-6 virus that had previously been deactivated by heat or ultraviolet light. Control larvae were not exposed to virus. Larval development time was shortest in control larvae and extended in larvae exposed to untreated virus. Covertly infected mosquitoes laid significantly fewer eggs, produced between 20 and 35% fewer progeny and had reduced longevity compared to other treatments. Wing length was shortest in mosquitoes exposed to heat-deactivated virus. Multivariate analysis of the same data identified fecundity and progeny production as the most influential variables in defining differences among treatments. Overall, viral infection resulted in a 34% decrease in the net reproductive rate (R0) of covertly infected mosquitoes, vs. only 5-17% decrease of R0 following treatments with deactivated virus, compared to controls. Sublethal effects of IIV-6 in Ae. aegypti appear to be mainly due to virus replication, rather than cytotoxic effects of the viral inoculum.

Update on the biology of Triatoma dimidiata Latreille (Hemiptera: Reduviidae) under laboratory conditions.

Feeding time, postfeeding defecation delay, and life cycle for each stage of a cohort of recently colonized Mexican Triatoma dimidiata were evaluated, and results were compared to existing published information on this species. Seventy-five nymphs (41.7%) completed a cycle with an average time from N-I to adult of 142 +/- 64 days. The average span in days for each stage was 20.2 for N-I, 17.9 for N-II, 10.1 for N-III, 43.6 for N-IV, and 55.1 for N-V. First-stage nymphs had the highest mean feeding time (25 min) and the longest postfeeding defecation delay (45 min). Differences among biological data from previous studies and the present study confirm the importance of conducting research on the behavior of the indigenous triatomine species from various countries.

Description and morphometric analysis of the eggs of Anopheles (Anopheles) vestitipennis (Diptera: Culcidae) from southern Mexico.

Light and scanning electron microscopy were used to compare the eggs of Anopheles vestitipennis Dyar & Knab females collected from human and animal baits in 9 villages of southern Mexico. An. vestitipennis eggs are boat-shaped, with lateral floats extending the length of the egg. Both the deck and dorsal surface are covered with hexagonal and pentagonal chorionic cells that contain round tubercles in the cell field. Crowns that enclose 3-5 lobed tubercles are present at both egg poles. By light microscopy, the mean length/width ratio of eggs of females caught at human bait were statistically different from those of females caught in horse-baited animal traps. In a regression tree model that included 19 egg attributes, the same egg groups could be separated by their posterior crown length/width ratio and mean anterior cell deck form factor (an index of their roundness). These findings support of the possible existence of 2 An. vestitipennis subpopulations with different host preferences in southern Mexico.

Gonotrophic cycle and survivorship of Anopheles vestitipennis (Diptera: Culicidae) in two different ecological areas of southern Mexico.

The duration of the gonotrophic cycle and survivorship of Anopheles vestitipennis Dyar & Knab was estimated in 2 malarious areas of Chiapas, Mexico: the Lacandon Forest and the Pacific Ocean Coastal Plain. Blood-engorged females held in an outdoor cage required 2.75 d for egg maturation, and 3.75 d for the duration of the gonotrophic cycle. Duration of the gonotrophic cycle also was estimated by parous-nulliparous dynamics for 20 consecutive days and autocorrelation time-series analysis, and by mark-recapture techniques. These methods depicted differences between the Lacandon Forest (3-d cycle) and the Coastal Plain (2-3 d cycles). Daily survival rates were estimated vertically and were generally higher in the Lacandon Forest (0.68) than in the Coastal Plain (0.45-0.58). The probability of mosquitoes surviving the sporogonic cycle was 10-100 times greater in the Lacandon Forest. The pregravid rate was 8.2%, and 29.3% of females with primary follicles beyond Christophers' stage III had traces of red blood in the gut. The 1st statistic indicated that 8.2% of females required > 1 blood meal for initial egg development, the 2nd statistic indicated that 29.3% of females take > 1 blood meal during a gonotrophic cycle. In summary, the enhanced vectorial role of this species is explained partially by high longevity and multiple blood-feeding habits.

Resistance management strategies in malaria vector mosquito control. Baseline data for a large-scale field trial against Anopheles albimanus in Mexico.

A high level of DDT resistance and low levels of resistance to organophosphorus, carbamate and pyrethroid insecticides were detected by discriminating dose assays in field populations of Anopheles albimanus in Chiapas, southern Mexico, prior to a large-scale resistance management project described by Hemingway et al. (1997). Biochemical assays showed that the DDT resistance was caused by elevated levels of glutathione S-transferase (GST) activity leading to increased rates of metabolism of DDT to DDE. The numbers of individuals with elevated GST and DDT resistance were well correlated, suggesting that this is the only major DDT resistance mechanism in this population. The carbamate resistance in this population is conferred by an altered acetylcholinesterase (AChE)-based resistance mechanism. The level of resistance observed in the bioassays correlates with the frequency of individuals homozygous for the altered AChE allele. This suggests that the level of resistance conferred by this mechanism in its heterozygous state is below the level of detection by the WHO carbamate discriminating dosage bioassay. The low levels of organophosphate (OP) and pyrethroid resistance could be conferred by either the elevated esterase or monooxygenase enzymes. The esterases were elevated only with the substrate pNPA, and are unlikely to be causing broad spectrum OP resistance. The altered AChE mechanism may also be contributing to the OP but not the pyrethroid resistance. Significant differences in resistance gene frequencies were obtained from the F1 mosquitoes resulting from adults obtained by different collection methods. This may be caused by different insecticide selection pressures on the insects immediately prior to collection, or may be an indication that the indoor- and outdoor-resting A. albimanus collections are not from a randomly mating single population. The underlying genetic variability of the populations is currently being investigated by molecular methods.

Colonization of Anopheles pseudopunctipennis from Mexico.

Two colonies of Anopheles pseudopunctipennis, Tapachula and Abasolo strains, were established under laboratory conditions with a thermoperiod (29 degrees C during the day; 24 degrees C during the night) and artificial dusk. To stimulate mating, a light beam from a flashlight was shone on the cage shortly after lights off. This procedure was repeated for the first 6 mosquito generations (parental to F6) and thereafter light stimulation was unnecessary for mating. The Tapachula colony has been maintained for 24 generations in 24 months, with insemination rates in females > 80% since the F3, and a monthly production of 30,000 pupae since the F7. Using the same procedure, the Abasolo colony from northeastern Mexico has been maintained for 13 generations in 14 months, with insemination rates of 26-52%.

Selective and conventional house-spraying of DDT and bendiocarb against Anopheles pseudopunctipennis in southern Mexico.

Indoor feeding behaviors and mortalities of Anopheles pseudopunctipennis females were evaluated following contact with selective (bands covering mosquitoes' preferred resting areas) and full applications of DDT and bendiocarb on indoor sprayable surfaces. The DDT residues provoked strong avoidance behavior. To a lesser degree, mosquitoes were also repelled by bendiocarb-sprayed surfaces. Because of strong irritancy/repellency, unfed mosquitoes were driven outdoors in proportionally higher numbers. The resting time on selectively or fully DDT-sprayed huts was greatly reduced in comparison to bendiocarb-sprayed huts. Although unfed mosquitoes tended to rest on non-DDT-sprayed surfaces in the selectively treated hut, the man-biting rate was similar with both types of treatments. Unfed mosquitoes were repelled less from selectively bendiocarb-treated surfaces. Similar reductions in postfed resting times were observed on all surfaces suggesting that once fed, mosquitoes rested on sprayed surfaces for shorter intervals of time. Engorged mosquitoes had normal resting behavior (pre- and postspray) within the range of preferred resting heights in both DDT- and bendiocarb-sprayed huts, but the proportion of mosquitoes fed in the DDT-treated huts was lower. Selective spraying of walls was as effective as spraying the complete walls with both insecticides, but DDT was more effective in reducing mosquito-human contact. These studies show that by more effectively targeting vector behavior, a cost-effective alternative to traditional control techniques can be achieved.

Influence of plant abundance on nectar feeding by Aedes aegypti (Diptera: Culicidae) in southern Mexico.

The availability of flowering plants affected the sugar feeding rates of female Aedes aegypti (L.) in 4 areas of a small city in southern Mexico. The proportion of mosquitoes containing sugar varied from 8 to 21% in 4 areas in direct relation to blooming plant abundance. Human density was similar in the 4 areas (range, 3.9-5.4 per house), whereas the number of flowering plants per house increased on the outskirts (range, 3.1-5.4 plants per house). Equal proportions of sugar positive females were nulliparous or parous, indicating similar sugar feeding at any age. In addition, nearly 60% of positive females were at the Christophers stage II, indicating a greater need for flight fuel during the early stages of egg development. We conclude that Ae. aegypti feeds frequently on nectar and that this activity is modulated by nectar availability.

Genetic differences among Anopheles vestitipennis subpopulations collected using different methods in Chiapas state, southern México.

Biting activity and population genetic studies of the malaria vector Anopheles vestitipennis were conducted in southern México. Three subpopulations were collected from 2 villages; 2 subpopulations were from the same village, one on human bait and one with an animal-baited trap; the third was collected from a cattle corral in the 2nd village (280 km away SSE). The anthropophilic subpopulation had steady activity with 61% of bites occurring before midnight, significantly different from those of the 2 zoophilic subpopulations, which had 78-82% of bites before midnight and 2 biting peaks, one at 1900-2100 h and the other at 0400-0500 h. Isozyme analysis (13 enzymes) of these subpopulations indicated that differences between the 2 sympatric subpopulations (D = 0.07), collected using 2 different methods, were greater than that between the 2 allopatric ones (D = 0.03). These studies suggest the existence of 2 genetically different subpopulations of An. vestitipennis with specific host preferences.

Low-volume application by mist-blower compared with conventional compression sprayer treatment of houses with residual pyrethroid to control the malaria vector Anopheles albimanus in Mexico.

Village-scale trials were carried out in southern Mexico to compare the efficacy of indoor-spraying of the pyrethroid insecticide lambda-cyhalothrin applied either as low-volume (LV) aqueous emulsion or as wettable-powder (WP) aqueous suspension for residual control of the principal coastal malaria vector Anopheles albimanus. Three indoor spray rounds were conducted at 3-month intervals using back-pack mist-blowers to apply lambda-cyhalothrin 12.5 mg a.i./m2 by LV, whereas the WP was applied by conventional compression sprayer at a mean rate of 26.5 mg a.i./m2. Both treatments caused mosquito mortality indoors and outdoors (collected inside house curtains) as a result of contact with treated surfaces before and after feeding, but had no significant impact on overall population density of An. albimanus resting indoors or assessed by human bait collections. Contact bioassays showed that WP and LV treatments with lambda-cyhalothrin were effective for 12-20 weeks (> 75% mortality) without causing excito-repellency. Compared to the WP treatment (8 houses/man/day), LV treatment (25 houses/man/day) was more than 3 times quicker per house, potentially saving 68% of labour costs. This is offset, however, by the much lower unit price of a compression sprayer (e.g. Hudson 'X-pert' at US$120) than a mist-blower (e.g. 'Super Jolly' at US$350), and higher running costs for LV applications. It was calculated, therefore, that LV becomes more economical than WP after 18.8 treatments/100 houses/10 men at equivalent rates of application, or after 7.6 spray rounds with half-rate LV applications.

Control of Anopheles albimanus mosquitos in southern Mexico by spraying their preferred indoor resting sites.

The traditional indoor spraying technique to control Anopheles albimanus mosquitos was compared with a selective method that targets their preferred resting sites in coastal villages of southern Mexico. We also determined whether mosquitos changed their preferred resting sites following insecticide applications. In the selective spraying approach, two 1-m wide horizontal swaths of bendiocarb insecticide were applied 0.75-1.75 m from the base of the walls and at 1-m upwards from the base of the inner roof at a target dose of 0.4 g/m2. A 2-year follow-up (1988-90), which included four insecticide treatments, showed that the selective spraying was as effective as full spraying for controlling A. albimanus. Also the mosquitos preferred resting sites that were not modified by the insecticide applications. Finally, the use of this techniques required 46% less time and cost 67% less than conventional spraying.

Remote sensing of tropical wetlands for malaria control in Chiapas, Mexico.

Malaria, transmitted by anopheline mosquitoes, remains a serious health problem in the tropics. Most malaria eradication efforts focus on control of anopheline vectors. These efforts include the NASA Di-Mod project, whose current goal is to integrate remote sensing, geographic information systems (GIS), and field research to predict anopheline mosquito population dynamics in the Pacific coastal plain of Chiapas, Mexico. Field studies demonstrate that high larval production of Anopheles albimanus, the principal malaria vector in the plain, can be linked to a small number of larval habitat-types, determined by larval sampling and cluster analysis of wetlands in the coastal plain. Analysis of wet and dry season Landsat Thematic Mapper (TM) satellite imagery identified 16 land cover units within an 185-km2 study area in the coastal zone. A hierarchical approach was used to link the larval habitat-types with the larger land cover units and make predictions of potential and actual low, medium, and high anopheline production. The TM-based map and GIS techniques were then used to predict differences in anopheline production at two villages, La Victoria and Efrain Gutierrez. La Victoria was predicted to have much higher Anopheles albimanus production, based upon a 2-10 times greater extent of medium- and high-producing land cover units in its vicinity. This difference between villages was independently supported by sampling (with light traps) of adults, which were 5-10 times more abundant in La Victoria.

Intradomiciliary behavior of Anopheles albimanus on the coastal plain of southern Mexico: implications for malaria control.

The postfeeding indoor resting behavior of Anopheles albimanus in experimental houses in southern México was investigated by using a mark-recapture procedure. The majority of mosquitoes rested inside houses after taking a blood meal indoors. There was a higher landing frequency on interior surfaces other than walls and roofs; however, mosquitoes rested for longer periods on these 2 surfaces. Successive landings on walls after short flights showed that mosquitoes gradually increased their mean landing height from 1.0 to 1.4 m. Similarly, mosquitoes resting at the base of inner roofs had a successive landing height range of about 0.5 m. Based on these observations and the potential for reduction of nearly 50% in the quantity of insecticide used and the time needed to apply it, village-scale studies involving the selective spraying of a 1-m-wide swath of insecticide on walls and on roofs are recommended in this area.

Indoor low-volume insecticide spray for the control of Anopheles albimanus in southern Mexico. Village-scale trials of bendiocarb, deltamethrin and cyfluthrin.

A comparative insecticide village-scale trial was carried out to determine the efficacy of low-volume (LV) indoor spray of bendiocarb, deltamethrin and cyfluthrin for the control of Anopheles albimanus in a coastal plain of southern Mexico. Low-volume spray was conducted with knapsack mist-blowers, giving an average discharge rate of 215 ml/min, which deposited droplets of 50-100 microns. Using this technique, 25 houses were treated/sprayman/day as compared with 8 houses/sprayman/day using conventional hand compression pumps. Indicators of LV mortality showed a residual activity of 10 wk as compared with 13 wk of activity for wettable powder (WP) (bendiocarb). When comparing expenditures, LV spray costs were 43% less when using bendiocarb, 83% less with deltamethrin and 74% less with cyfluthrin with respect to the conventional WP spray of the same insecticides. Low-volume spray time was reduced by one-third, with respect to WP. Malaria incidence was reduced by 53 and 56%, respectively, in the bendiocarb LV and deltamethrin LV treated villages as compared with the untreated village.

Anopheles albimanus (Diptera: Culicidae) host selection patterns in three ecological areas of the coastal plains of Chiapas, southern Mexico.

The host-feeding patterns of Anopheles albimanus Wiedemann were described and the effect of host availability on these patterns was assessed in three different ecological areas of coastal Chiapas, Mexico. Resting mosquitoes were collected indoors and outdoors during rainy seasons. A 20% sample of blood-fed mosquitoes was tested to determine the source of the blood meal using an ELISA. The unweighted human blood index (HBI) of An. albimanus in the three areas ranged from 0.11 to 0.21. This mosquito species fed more frequently on bovines than on any other host, but the forage ratio indicated that there was also a high preference for equines. Some females tended to rest or complete their gonotrophic cycle indoors after feeding on animals, but females also fed on man and rested outdoors. Host availability and ecological conditions appeared to be responsible for differences observed in the HBI among areas.

[Effectiveness of a carbamate insecticide as a household low-volume spray for malaria control]. / Efectividad de un insecticida carbamato en rociado intradomiciliar a bajo volumen para el control del paludismo.

A comparative regional-scale evaluation of the epidemiological impact of low volume (LV) spray of bendiocarb and the conventional spray of DDT against malaria in an endemic area of northern Chiapas was carried out. Three Anopheline species were found: Anopheles pseudopunctipennis, An. albinanus and An. argyritarsis. The most prevalent was An. pseudopunctipennis, a species we suspect may be involved in the transmission of most malaria cases. This species showed high levels of resistance to DDT. However, this insecticide had a long residual effect in wall bioassays, with mortalities > or = 95 per cent for up to 21 weeks in wood, sticks and plaster. Susceptibility to bendiocarb was total, and mortality to LV bendiocarb was > or = 75 per cent for up to 16 weeks in wood, sticks and straw. Very low numbers of mosquitoes were found throughout the evaluation, although malaria transmission continued in control villages (sprayed with DDT), as well as treated villages (sprayed with LV bendiocarb). No plasmodium infected mosquitoes were found, perhaps due to a very small sample size. The LV spray methodology was found to be 1.7 times more effective than conventional spraying in reducing malaria incidence. A net reduction of 1.6 times in insecticide application time was also found, which would allow spraying at the right time, especially when urgent control measures have to be applied, such as in malaria outbreaks. Finally, the new methodology costs 2.2 times more than the conventional ddt spraying, but if the potential of using spray workers in other activities is considered, costs would be comparable to those of DDT spraying.

Gonotrophic cycle and survivorship of Anopheles albimanus (Diptera: Culicidae) in southern Mexico.

The length of the gonotrophic cycle of Anopheles albimanus was estimated by 12 mark-release-recapture studies conducted in corrals in southern Mexico from 1987 to 1990. The initial set of three mark-release studies indicated that the gonotrophic cycle takes at least 4 d based on the day when gravid mosquitoes were first recaptured. However, in later experiments, mosquitoes recaptured seeking hosts at 48 h after release were in Sella's and Christophers' stages I and II, but parity rates had nearly doubled, indicating that eggs may have developed in less than 48 h and that mosquitoes returned to refeed immediately following oviposition. Two gonotrophic cycles probably exist, one of 48 h dominated by parous mosquitoes and one of 4 d comprised of nulliparous, pregravid mosquitoes. Daily survivorship was estimated by regression from the decrease in the daily recapture rate (0.46-0.68) was less than that estimated by the parity rates (0.67-0.69), with no important differences found between wet and dry seasons. Estimates of the probability of a mosquito living long enough to transmit malaria were 2% (range, 1.8-2.5%).

Age structure and abundance levels in the entomological evaluation of an insecticide used in the control of Anopheles albimanus in southern Mexico.

Applications of bendiocarb produced a high insecticidal residual effect lasting up to 3 months on the most common indoor house surfaces. No significant decreases in mosquito man-biting rate levels were observed between treated and untreated villages. It was shown that almost equal proportions of intra- and peridomicillary mosquitoes came into contact with the insecticide, indicating that mosquitoes commonly enter houses, rest on treated surfaces and return to bite both indoors and outdoors. Although the insecticide was found to have a significant effect on the percentage parity (interpreted as abundance of older individuals) of intra- and peridomicillary Anopheles albimanus mosquitoes, parity recovered and continued a normal cyclic pattern that appeared to be dependent on relative abundance. The proximity of a treated village to an untreated village (1.2 km) can affect the age structure of mosquito populations through shared common resting and breeding sites.