Filiberto Reyes-Villanueva

Transmission of Beauveria bassiana from male to female Aedes aegypti mosquitoes

BackgroundResistance to chemical insecticides plus high morbidity rates have lead to rising interest in fungi as candidates for biocontrol agents of mosquito vectors. In most studies fungal infections have been induced by exposure of mosquitoes to various surfaces treated with conidia. In the present study eight Mexican strains of Beauveria bassiana were assessed against Aedes aegypti by direct exposure of females to 6 × 108 conidia ml -1 on a filter paper, afterwards, the transmission of the least and most virulent isolates was evaluated by mating behavior from virgin, fungus-contaminated male to females, to examine this ethological pattern as a new approach to deliver conidia against the dengue vector.MethodsIn an exposure chamber with a filter paper impregnated with 6 × 108 conidia ml -1 of the least and most virulent strains of B. bassiana, 6-8 day old males of A. aegypti were exposed for 48 hours, and then transferred individually (each one was a replicate) to another chamber and confined with twenty healthy females of the same age. Clean males were used in controls. Survival, infection by true mating (insemination) or by mating attempts (no insemination) and fecundity were daily registered until the death of last female. Data analysis was conducted with proc glm for unbalanced experiments and means were separated with the Ryan test with SAS.ResultsAll strains were highly virulent with LT50 ranging from 2.70 (± 0.29) to 5.33 (± 0.53) days. However the most (Bb-CBG2) and least virulent (Bb-CBG4) isolates were also transmitted by mating behavior; both killed 78-90% of females in 15 days after being confined with males that had previously been exposed for 48 hours to fungi. Of these mortality rates, 23 and 38% respectively, were infections acquired by copulations where insemination occurred. The LT50 for sexually-infected females were 7.92 (± 0.46) and 8.82 (± 0.45) days for both strains, while the one in control was 13.92 (± 0.58). Likewise, fecundity decreased by 95% and 60% for both Bb-CBG2 and Bb-CBG4 isolates in comparison with control. The role of mating attempts in this delivery procedure of B. bassiana is discussed.ConclusionsThis is the first report about transmission of B. bassiana by mating behavior from virgin, fungus-contaminated males to females in A. aegypti. Fungal infections acquired by this route (autodissemination) infringed high mortality rates (90%) in mated or approached females. However, prior to releasing virgin, fungus-contaminated males to spread B. basasiana among females of A. aegypti, this novel alternative needs further investigations.

Development of a novel trap for the collection of black flies of the Simulium ochraceum complex.

Background Human landing collections are currently the standard method for collecting onchocerciasis vectors in Africa and Latin America. As part of the efforts to develop a trap to replace human landing collections for the monitoring and surveillance of onchocerciasis transmission, comprehensive evaluations of several trap types were conducted to assess their ability to collect Simulium ochraceum sensu lato, one of the principal vectors of Onchocerca volvulus in Latin America. Methodology/Principal Findings Diverse trap designs with numerous modifications and bait variations were evaluated for their abilities to collect S. Ochraceum s.l. females. These traps targeted mostly host seeking flies. A novel trap dubbed the “Esperanza window trap” showed particular promise over other designs. When baited with CO2 and BG-lure (a synthetic blend of human odor components) a pair of Esperanza window traps collected numbers of S. Ochraceum s.l. females similar to those collected by a team of vector collectors. Conclusions/Significance The Esperanza window trap, when baited with chemical lures and CO2 can be used to collect epidemiologically significant numbers of Simulium ochraceum s.l., potentially serving as a replacement for human landing collections for evaluation of the transmission of O. volvulus.

Dissemination of Metarhizium anisopliae of low and high virulence by mating behavior in Aedes aegypti.

BackgroundDengue is a viral disease transmitted by Aedes mosquitoes. It is a threat for public health worldwide and its primary vector Aedes aegypti is becoming resistant to chemical insecticides. These factors have encouraged studies to evaluate entomopathogenic fungi against the vector. Here we evaluated mortality, infection, insemination and fecundity rates in A. aegypti females after infection by autodissemination with two Mexican strains of Metarhizium anisopliae.MethodsTwo M. anisopliae strains were tested: The Ma-CBG-1 least virulent (lv), and the Ma-CBG-2 highly virulent (hv) strain. The lv was tested as non mosquito-passed (NMP), and mosquito-passed (MP), while the hv was examined only as MP version, therefore including the control four treatments were used. In the first bioassay virulence of fungal strains towards female mosquitoes was determined by indirect exposure for 48 hours to conidia-impregnated paper. In the second bioassay autodissemination of fungal conidia from fungus-contaminated males to females was evaluated. Daily mortality allowed computation of survival curves and calculation of the LT50 by the Kaplan-Meier model. All combinations of fungal sporulation and mating insemination across the four treatments were analyzed by χ2. The mean fecundity was analyzed by ANOVA and means contrasted with the Ryan test.ResultsIndirect exposure to conidia allowed a faster rate of mortality, but exposure to a fungal-contaminated male was also an effective method of infecting female mosquitoes. All females confined with the hv strain-contaminated male died in fifteen days with a LT50 of 7.57 (± 0.45) where the control was 24.82 (± 0.92). For the lv strain, it was possible to increase fungal virulence by passing the strain through mosquitoes. 85% of females exposed to hv-contaminated males became infected and of them just 10% were inseminated; control insemination was 46%. The hv strain reduced fecundity by up to 99%, and the lv strain caused a 40% reduction in fecundity.ConclusionsThe hv isolate infringed a high mortality, allowed a low rate of insemination, and reduced fecundity to nearly zero in females confined with a fungus-contaminated male. This pathogenic impact exerted through sexual transmission makes the hv strain of M. anisopliae worthy of further research.

Vectorial Capacity of Aedes aegypti for Dengue Virus Type 2 Is Reduced with Co-infection of Metarhizium anisopliae

Background Aedes aegypti, is the major dengue vector and a worldwide public health threat combated basically by chemical insecticides. In this study, the vectorial competence of Ae. aegypti co-infected with a mildly virulent Metarhizium anisopliae and fed with blood infected with the DENV-2 virus, was examined. Methodology/Principal Findings The study encompassed three bioassays (B). In B1 the median lethal time (LT50) of Ae. aegypti exposed to M. anisopliae was determined in four treatments: co-infected (CI), single-fungus infection (SF), single-virus infection (SV) and control (C). In B2, the mortality and viral infection rate in midgut and in head were registered in fifty females of CI and in SV. In B3, the same treatments as in B1 but with females separated individually were tested to evaluate the effect on fecundity and gonotrophic cycle length. Survival in CI and SF females was 70% shorter than the one of those in SV and control. Overall viral infection rate in CI and SV were 76 and 84% but the mortality at day six post-infection was 78% (54% infected) and 6% respectively. Survivors with virus in head at day seven post-infection were 12 and 64% in both CI and SV mosquitoes. Fecundity and gonotrophic cycle length were reduced in 52 and 40% in CI compared to the ones in control. Conclusion/Significance Fungus-induced mortality for the CI group was 78%. Of the survivors, 12% (6/50) could potentially transmit DENV-2, as opposed to 64% (32/50) of the SV group, meaning a 5-fold reduction in the number of infective mosquitoes. This is the first report on a fungus that reduces the vectorial capacity of Ae. aegypti infected with the DENV-2 virus.

Mosquito Records From Mexico: The Mosquitoes (Diptera: Culicidae) of Tamaulipas State

ABSTRACT To document the diversity and distribution of mosquito species inhabiting the Mexican state of Tamaulipas, collection trips were conducted to all physiographic regions (Grand Northamerican Plains, Coastal Plain of North Gulf, and Sierra Madre Oriental) and subregions across the state. Additionally, we re-examined mosquito specimens in two Mexican entomological collections: the Collection of Insects and Mites of Medical Importance and the Collection of Arthropods of Medical Importance. In total, 3,931 specimens were collected. These represent the two Culicidae subfamilies Anophelinae and Culicinae, 10 tribes, 17 genera, 27 subgenera, 80 named species, and 2 undescribed species. Of these, 3 tribes, 6 genera, 7 subgenera, and 20 species are new records for the mosquito fauna of Tamaulipas. Fourteen species recorded in the historical records were not found in collections made for this study. Taxonomic notes, new distribution limits, and comments about the medical importance of some of the species collected are reported.

Biological Control of Dengue Vectors

Biological control is the deliberate use of natural enemies to reduce the number of pest organisms. It comprises methods that have gained acceptance for controlling nuisance arthropods partly due to the emergence of insecticide resistance and also because people have become more aware about the need to limit environmental pollution. In the case of arthropod-borne disease vectors, biological control is a potentially effective strategy for regulating and preventing transmission of diseases such as dengue, malaria and lymphatic filariasis, amongst others. Dengue is an arbovirus transmitted by species of Aedes mosquitoes. Aedes aegypti and Aedes albopictus are the primary and secondary worldwide vectors; they breed in peridomestic man-made water containers, and their control is the most effective way to reduce the viral transmission.

The esperanza window trap reduces the human biting rate of Simulium ochraceum s.l. in formerly onchocerciasis endemic foci in Southern Mexico.

Background The Esperanza Window Trap (EWT) baited with CO2 and human sweat compounds is attractive to Simulium ochraceum s.l., the primary vector of Onchocerca volvulus in the historically largest endemic foci in México and Guatemala. Methodology/Principal findings The ability of the EWT to locally reduce numbers of questing S. ochraceum s.l. was evaluated in two formerly onchocerciasis endemic communities in Southern México. At each community, two EWTs were placed in or near a school or household and flies were collected sequentially for a total of 10 days. Black fly collections were then carried out for an additional 10 days in the absence of the EWTs. Flies were also collected outside the dwellings to control for variations in the local fly populations. When the EWTs were present, there was a significant reduction in the human biting rate at both the household and school locations at collection sites, with a greater effect observed in the schools. Conclusions/Significance These results indicate that the EWTs not only have potential as a black fly monitoring tool but may be used for reducing personal exposure to fly bites in Mesoamerica.

Copulation Activity, Sperm Production and Conidia Transfer in Aedes aegypti Males Contaminated by Metarhizium anisopliae: A Biological Control Prospect.

Background Dengue is the most prevalent arboviral disease transmitted by Aedes aegypti worldwide, whose chemical control is difficult, expensive, and of inconsistent efficacy. Releases of Metarhizium anisopliae—exposed Ae. aegypti males to disseminate conidia among female mosquitoes by mating represents a promising biological control approach against this important vector. A better understanding of fungus virulence and impact on reproductive parameters of Ae. aegypti, is need before testing auto-dissemination strategies. Methodology/Principal Findings Mortality, mating competitiveness, sperm production, and the capacity to auto-disseminate the fungus to females up to the 5thcopulation, were compared between Aedes aegypti males exposed to 5.96 x 107 conidia per cm2 of M. anisopliae and uninfected males. Half (50%) of fungus-exposed males (FEMs) died within the first 4 days post-exposure (PE). FEMs required 34% more time to successively copulate with 5 females (165 ± 3 minutes) than uninfected males (109 ± 3 minutes). Additionally, fungus infection reduced the sperm production by 87% at 5 days PE. Some beneficial impacts were observed, FEMs were able to successfully compete with uninfected males in cages, inseminating an equivalent number of females (about 25%). Under semi-field conditions, the ability of FEMs to search for and inseminate females was also equivalent to uninfected males (both inseminating about 40% females); but for the remaining females that were not inseminated, evidence of tarsal contact (transfer of fluorescent dust) was significantly greater in FEMs compared to controls. The estimated conidia load of a female exposed on the 5th copulation was 5,200 mL-1 which was sufficient to cause mortality. Conclusion/Significance Our study is the first to demonstrate auto-dissemination of M. anisopliae through transfer of fungus from males to female Ae. aegypti during mating under semi-field conditions. Our results suggest that auto-dissemination studies using releases of FEMs inside households could successfully infect wild Ae. aegypti females, providing another viable biological control tool for this important the dengue vector.

Twenty-three years after the first record of Aedes albopictus in Nigeria : its current distribution and potential epidemiological implications

Aedes albopictus (Skuse) was first recorded in Nigeria in 1991 during a post epidemiological surveillance of yellow fever epidemics in Delta State. In this report, we present a map of the current distribution of Ae. albopictus and its possible epidemiological implications based on accessible records of mosquito fauna in Nigeria using electronic e-journal searching engines. We also report the results of a field study on population dominance between Aedes aegypti and Ae. albopictus in a woodland habitat in southwestern Nigeria between March and November 2002. Of over 100 accessible papers on mosquito fauna in Nigeria, only 18 have reported information on Ae. albopictus. Literature shows that Ae. albopictus became well established in southeastern Nigeria and then spread to other southern regions. There was no information about the species in northern Nigeria except Kano where a record of its breeding was reported. Data showed dominance of Ae. albopictus over Ae. aegypti in southeastern Nigeria but the reverse was observed during a field investigation at Ikenne farm settlement in southwestern Nigeria. Aedes albopictus was perhaps involved in the transmission of arboviral infections and Bancroftian filariasis in southern Nigeria. Data show that Ae. albopictus is well established in southern Nigeria but not in arid zones of the country. Its establishment and role as a vector of arboviral infections and Bancroftian filariasis in southern Nigeria should be a source of concern. The present report therefore serves as a baseline map for future epidemiological surveillance and the understanding of ecological distribution of Ae. albopictus in the country.

First Record of Culiseta melanura in Mexico, with ADditional Mexican Records for Aedes sollicitans

Abstract Larvae of Culiseta melanura were collected during June 2001 and January 2002 in Nuevo Leon State, Mexico. This is the first record of this species in Mexico. Female and male adults of Aedes sollicitans were collected during May 1992 and June 1994 in Tamaulipas State and Nuevo Leon State, respectively; adult females of Ae. sollicitans were collected also during 1996 from a resting place in Oaxaca State. These records for Ae. sollicitans supplement previous Mexican records of the species. All specimens were identified through examination of adult females, male genitalia, and associated larval exuviae when available.