Traditional and Geometric Morphometry Analyses of Lutzomyia cruciata (Diptera: Psychodidae: Phlebotominae) Populations of Chiapas, Mexico.

The presence of Lutzomyia (Tricholateralis) cruciata (Coquillett 1907) species complex has been suggested by morphological analysis of eggs and genetic studies of females. The present work aimed to compare the diversity in morphology of four populations of Lu. cruciata from the Coast of Chiapas, Mexico, using traditional (TM) and geometric (GM) methods. Several morphological characteristics that were analyzed provided consistency to differentiate at least, three populations of Lu. cruciata. Both methods were effective to detect morphological differences associated with the geographical sites of capture. In both sexes, three and four groups were detected by TM and GM, respectively. These results suggest marked morphological differences in both sexes of Lu. cruciata that make these methods potentially useful to identify the geographical origin of any specimen of this species captured in the study region. Although the results produced by both methods are coincident, geometric morphometrics turned out to be most advantageous with respect to traditional morphometry, since the latter requires more time and effort. The consistency of our results shows that the variability of environmental conditions on the coast of Chiapas determines a high degree of phenotypic plasticity in Lu. cruciata, with the possibility of prezygotic isolation and the formation of species complex.

Spinosad, a naturally derived insecticide, for control of Aedes aegypti (Diptera: Culicidae): efficacy, persistence, and elicited oviposition response.

The naturally derived insecticide spinosad is a reduced-risk material that is neurotoxic to Diptera. The 24-h 50% lethal concentration by laboratory bioassay in third instars of Aedes aegypti (L.) (Diptera: Culicidae) (Rockefeller strain) was estimated at 0.026 ppm. Two identical field trials were performed in an urban cemetery in southern Mexico during the dry and wet seasons. Water containers treated with 1 or 5 ppm spinosad suspension concentrate (Tracer, Dow Agrosciences) were as effective in preventing the development of Aedes spp. (mostly Ae. aegypti) as temephos granules during both trials, whereas the bacterial insecticide VectoBac 12AS performed poorly. The half-life of aqueous solutions of spinosad (10 ppm) placed in a warm sunny location was 2.1 d, compared with 24.5 d for solutions in a shaded location. Spinosad, temephos, and VectoBac were not repellent to gravid Ae. aegypti at the concentrations tested, and no ovicidal properties were observed. The 24-h survival of neonate larvae but was reduced by 94-100% in the presence of residues carried over from the spinosad treatments, but it was not affected by residues of temephos or VectoBac. The toxicological properties of spinosad, combined with its favorable environmental profile, should encourage the detailed evaluation of spinosad as a mosquito larvicide in domestic and urban environments.

Efficiency of two larval diets for mass-rearing of the mosquito Aedes aegypti.

Aedes aegypti is a major vector of arboviruses that may be controlled on an area-wide basis using the sterile insect technique (SIT). Larval diet is a major factor in mass-rearing for SIT programs. We compared dietary effects on immature development and adult fitness-related characteristics for an International Atomic Energy Agency (IAEA) diet, developed for rearing Ae. albopictus, and a standardized laboratory rodent diet (LRD), under a 14:10 h (light:dark) photoperiod ("light" treatment) or continuous darkness during larval rearing. Larval development was generally fastest in the IAEA diet, likely reflecting the high protein and lipid content of this diet. The proportion of larvae that survived to pupation or to adult emergence did not differ significantly between diets or light treatments. Insects from the LRD-dark treatment produced the highest proportion of male pupae (93% at 24 h after the beginning of pupation) whereas adult sex ratio from the IAEA diet tended to be more male-biased than that of the LRD diet. Adult longevity did not differ significantly with larval diet or light conditions, irrespective of sex. In other aspects the LRD diet generally performed best. Adult males from the LRD diet were significantly larger than those from the IAEA diet, irrespective of light treatment. Females from the LRD diet had ~25% higher fecundity and ~8% higher egg fertility compared to those from the IAEA diet. Adult flight ability did not differ between larval diets, and males had a similar number of copulations with wild females, irrespective of larval diet. The LRD diet had lower protein and fat content but a higher carbohydrate and energetic content than the IAEA diet. We conclude that the LRD diet is a low-cost standardized diet that is likely to be suitable for mass-rearing of Ae. aegypti for area-wide SIT-based vector control.

A comparison of techniques for detecting Invertebrate iridescent virus 6.

The aim of this study was to compare the sensitivity and precision of various methods for the detection and quantification of Invertebrate iridescent virus 6 (IIV-6) (Iridoviridae) isolated from a the stem-boring moth Chilo suppressalis, and to apply these techniques to the detection of covert infections in the wax moth, Galleria mellonella. The relationship between the virus concentration and absorbance at 260 nm was linear over the range of 1.6 x 10(9)-5.6 x 10(10) particles/ml. TCID(50) assays using 12 different cell lines indicated that two Drosophila lines, DL2 and DR1, had the highest susceptibility whereas cell lines from Aedes albopictus and Plutella xylostella were four orders of magnitude less sensitive. TCID(50) values for IIV-6 in Spodoptera frugiperda Sf9 cells gave the particle-infectivity ratios of 15-64 virus particles/IU. An insect bioassay involved injecting doses of 1-100 IIV-6 particles into the third instar G. mellonella larvae. The prevalence of patent infection was 20-26% at a dose of 1 particle per larva rising to 86-92% at 10 particles and 100% at doses of 50 or 100 particles. Of the insects that survived to adulthood, between 5.8 and 75% caused patent infections when injected into G. mellonella larvae, indicating that they were covertly infected. A PCR technique resulted in 95% detection at 1000 virus particles per insect. Of the insects that proved positive for covert infection by insect bioassay, 41% also proved positive by PCR analysis. It is concluded that the G. mellonella bioassay is highly reliable for detection of doses of 10 particles or more and for determining the relative activity of IIV-6 preparations at doses as low as 1 particle per insect. PCR had a slightly lower sensitivity followed by the insect cell culture assay.

Persistence of invertebrate iridescent virus 6 in tropical artificial aquatic environments. Brief report.

The rate of loss of activity of invertebrate iridescent virus 6 (IIV-6, family Iridoviridae) was determined in two artificial aquatic habitats in southern Mexico, using a sensitive insect bioassay technique. IIV-6 placed in trays of water in direct sunlight suffered rapid loss of activity (99.99% reduction) over a period of 36 h, during which water temperatures fluctuated between 24 and 41 degrees C. No significant deactivation occurred during the hours of darkness. In contrast, IIV-6 placed in trays of water in the shade lost 97% of original activity over a 60 h period, during which water temperatures fluctuated from 24 to 31 degrees C. Longitudinal analysis involving mixed effects models of time (shade) and cumulative exposure to ultraviolet radiation (sunlight) indicated that the rate of deactivation was best described by third order polynomial equations in both cases. We conclude that the likelihood of transmission of IIVs in aquatic habitats will be mediated by the intensity of UV radiation and water temperature.

The naturally derived insecticide spinosad is highly toxic to Aedes and Anopheles mosquito larvae.

Spinosad is a naturally derived biorational insecticide with an environmentally favourable toxicity profile, so we investigated its potency against mosquito larvae (Diptera: Culicidae). By laboratory bioassays of a suspension concentrate formulation of spinosad (Tracer), the 24 h lethal concentration (LC50) against Aedes aegypti (L.) third and fourth instars was estimated at 0.025 p.p.m. following logit regression. The concentration-mortality response of third- and fourth-instar Anopheles albimanus Weidemann did not conform to a logit model. The LC50 value of spinosad in Anopheles albimanus was 0.024 p.p.m. by quadratic linear regression. A field trial in southern Mexico demonstrated that spinosad 1 p.p.m. compared with the standard temephos (Abate) 1% granules 100 g/m3 water prevented Ae. aegypti breeding in plastic containers of water for 8 weeks; at 10 p.p.m. spinosad prevented breeding for > 22 weeks. In another field trial, spinosad at 5 p.p.m. and temephos both completely eliminated reproduction of Ae. aegypti for 13 weeks. In contrast, the bacterial insecticide Bacillus thuringiensis var. israelensis (Bti, Vectobac) AS) performed poorly with just 2 weeks of complete inhibition of Ae. aegypti breeding. Spinosad also effectively prevented breeding of Culex mosquitoes and chironomids in both trials to a degree similar to that of temephos. We conclude that spinosad merits evaluation as a replacement for organophosphate or Bti treatment of domestic water tanks in Mesoamerica. We also predict that spinosad is likely to be an effective larvicide for treatment of mosquito breeding sites.

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.