Why is Aedes aegypti Linnaeus so Successful as a Species?

Diseases transmitted by mosquitoes impose enormous burden towards human morbidity and mortality. Over the last three decades, Brazil has suffered from severe Dengue epidemics. In September 2014, this situation is further complicated by the introduction of two other viruses, Zika and Chikungunya, placing Brazil in a triple epidemic. In this article, we discuss the biology of Aedes aegypti Linnaeus, and the principal initiatives currently used to control mosquito populations and the diseases they transmit. Aedes aegypti has broad global distribution and is involved in the transmission of various arboviral diseases such as Dengue, Zika, and Chikungunya. Several factors contribute to the success of the species, particularly behavioral plasticity, rapid development, desiccation-resistant eggs, resistance to the principle insecticide classes currently available on the market, preference for the urban environment, and proximity to humans. Vector control programs are the best way to reduce the burden of mosquito-borne diseases. Chemical control is most commonly used in recent times, and unfortunately, the results have not been satisfactory but instead, there is increased vector dispersal and, subsequently, the spread of disease epidemics. Investigations of alternative control methods such as release of Wolbachia-infected mosquitoes for blocking vector-borne pathogens, release of transgenic mosquitoes carrying a lethal gene for offspring, and the use of insecticide-dispersing mosquitoes are under way in Brazil, and some have shown promising results. Special emphasis should be placed on integrated management of all available tactics, so as to maximize efforts towards mosquito control. Finally, we emphasize that continuous actions and community participation control initiatives are critically important for success.

The transcriptome of the mosquito Aedes fluviatilis (Diptera: Culicidae), and transcriptional changes associated with its native Wolbachia infection.

BACKGROUND: Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Stable infections of Wolbachia in mosquitoes can inhibit infection with medically important pathogens such as dengue virus and malaria-causing Plasmodium parasites. However, some native Wolbachia strains can enhance infection with certain pathogens, as is the case for the mosquito Aedes fluviatilis, where infection with Plasmodium gallinaceum is enhanced by the native wFlu Wolbachia strain. To better understand the biological interactions between mosquitoes and native Wolbachia infections, and to investigate the process of pathogen enhancement, we used RNA-Seq to generate the transcriptome of Ae. fluviatilis with and without Wolbachia infection. RESULTS: In total, we generated 22,280,160 Illumina paired-end reads from Wolbachia-infected and uninfected mosquitoes, and used these to make a de novo transcriptome assembly, resulting in 58,013 contigs with a median sequence length of 443 bp and an N50 of 2454 bp. Contigs were annotated through local alignments using BlastX, and associated with both gene ontology and KEGG orthology terms. Through baySeq, we identified 159 contigs that were significantly upregulated due to Wolbachia infection, and 98 that were downregulated. Critically, we saw no changes to Toll or IMD immune gene transcription, but did see evidence that wFlu infection altered the expression of several bacterial recognition genes, and immune-related genes that could influence Plasmodium infection. wFlu infection also had a widespread effect on a number of host physiological processes including protein, carbohydrate and lipid metabolism, and oxidative stress. We then compared our data set with transcriptomic data for other Wolbachia infections in Aedes aegypti, and identified a core set of 15 gene groups associated with Wolbachia infection in mosquitoes. CONCLUSIONS: While the scale of transcriptional changes associated with wFlu infection might be small, the scope is rather large, which confirms that native Wolbachia infections maintain intricate molecular relationships with their mosquito hosts even after lengthy periods of co-evolution. We have also identified several potential means through which wFlu infection might influence Plasmodium infection in Ae. fluviatilis, and these genes should form the basis of future investigation into the enhancement of Plasmodium by Wolbachia.

Broader prevalence of Wolbachia in insects including potential human disease vectors.

Wolbachia are intracellular, maternally transmitted bacteria considered the most abundant endosymbionts found in arthropods. They reproductively manipulate their host in order to increase their chances of being transmitted to the offspring, and currently are being used as a tool to control vector-borne diseases. Studies on distribution of Wolbachia among its arthropod hosts are important both for better understanding why this bacterium is so common, as well as for its potential use as a biological control agent. Here, we studied the incidence of Wolbachia in a broad range of insect species, collected from different regions of Brazil, using three genetic markers (16S rRNA, wsp and ftsZ), which varied in terms of their sensitivity to detect this bacterium. The overall incidence of Wolbachia among species belonging to 58 families and 14 orders was 61.9%. The most common positive insect orders were Coleoptera, Diptera, Hemiptera and Hymenoptera, with Diptera and Hemiptera having the highest numbers of Wolbachia-positive families. They included potential human disease vectors whose infection status has never been reported before. Our study further shows the importance of using quantitative polymerase chain reaction for high-throughput and sensitive Wolbachia screening.

The wMel Wolbachia strain blocks dengue and invades caged Aedes aegypti populations.

Dengue fever is the most important mosquito-borne viral disease of humans with more than 50 million cases estimated annually in more than 100 countries. Disturbingly, the geographic range of dengue is currently expanding and the severity of outbreaks is increasing. Control options for dengue are very limited and currently focus on reducing population abundance of the major mosquito vector, Aedes aegypti. These strategies are failing to reduce dengue incidence in tropical communities and there is an urgent need for effective alternatives. It has been proposed that endosymbiotic bacterial Wolbachia infections of insects might be used in novel strategies for dengue control. For example, the wMelPop-CLA Wolbachia strain reduces the lifespan of adult A. aegypti mosquitoes in stably transinfected lines. This life-shortening phenotype was predicted to reduce the potential for dengue transmission. The recent discovery that several Wolbachia infections, including wMelPop-CLA, can also directly influence the susceptibility of insects to infection with a range of insect and human pathogens has markedly changed the potential for Wolbachia infections to control human diseases. Here we describe the successful transinfection of A. aegypti with the avirulent wMel strain of Wolbachia, which induces the reproductive phenotype cytoplasmic incompatibility with minimal apparent fitness costs and high maternal transmission, providing optimal phenotypic effects for invasion. Under semi-field conditions, the wMel strain increased from an initial starting frequency of 0.65 to near fixation within a few generations, invading A. aegypti populations at an accelerated rate relative to trials with the wMelPop-CLA strain. We also show that wMel and wMelPop-CLA strains block transmission of dengue serotype 2 (DENV-2) in A. aegypti, forming the basis of a practical approach to dengue suppression.

Improved accuracy of the transcriptional profiling method of age grading in Aedes aegypti mosquitoes under laboratory and semi-field cage conditions and in the presence of Wolbachia infection.

Transcriptional profiling is an effective method of predicting age in the mosquito Aedes aegypti in the laboratory, however, its effectiveness is limited to younger mosquitoes. To address this we used a microarray to identify new gene candidates that show significant expression changes in older mosquitoes. These genes were then used to create a revised model, which upon evaluation in both laboratory and semi-field conditions, proved to have improved accuracy overall and for older mosquitoes. In association with the development of symbiont-based control strategies for Ae. aegypti, we also tested the model's accuracy for Wolbachia-infected mosquitoes and found no decline in performance. Our findings suggest that the new model is a robust and powerful tool for age determination in Australian Ae. aegypti populations.

Expression of a mutated phospholipase A2 in transgenic Aedes fluviatilis mosquitoes impacts Plasmodium gallinaceum development.

The genetic manipulation of mosquito vectors is an alternative strategy in the fight against malaria. It was previously shown that bee venom phospholipase A2 (PLA2) inhibits ookinete invasion of the mosquito midgut although mosquito fitness was reduced. To maintain the PLA2 blocking ability without compromising mosquito biology, we mutated the protein-coding sequence to inactivate the enzyme while maintaining the protein's structure. DNA encoding the mutated PLA2 (mPLA2) was placed downstream of a mosquito midgut-specific promoter (Anopheles gambiae peritrophin protein 1 promoter, AgPer1) and this construct used to transform Aedes fluviatilis mosquitoes. Four different transgenic lines were obtained and characterized and all lines significantly inhibited Plasmodium gallinaceum oocyst development (up to 68% fewer oocysts). No fitness cost was observed when this mosquito species expressed the mPLA2.

Biomphalaria tenagophila/Schistosoma mansoni interaction: premises for a new approach to biological control of schistosomiasis.

Biomphalaria tenagophila is very important for schistosomiasis transmission in Brazil. However its mechanisms of interaction with Schistosoma mansoni are still scantly studied. Since this snail displays strains highly susceptible or completely resistant to the parasite infection, the knowledge of that would be a useful tool to understand the mechanism of snail resistance. Particularly, the Taim strain consistently shows absolute resistance against the trematode, and this resistance is a dominant character. A multidisciplinary research group was created aiming at studying B. tenagophila/S. mansoni interaction. The possibility for applying the knowledge acquired to obtain a biological model for the control of S. mansoni transmission in endemic areas is discussed.

Biomphalaria tenagophila/Schistosoma mansoni interaction: premises for a new approach to biological control of schistosomiasis

Biomphalaria tenagophila is very important for schistosomiasis transmission in Brazil. However its mechanisms of interaction with Schistosoma mansoni are still scantly studied. Since this snail displays strains highly susceptible or completely resistant to the parasite infection, the knowledge of that would be a useful tool to understand the mechanism of snail resistance. Particularly, the Taim strain consistently shows absolute resistance against the trematode, and this resistance is a dominant character. A multidisciplinary research group was created aiming at studying B. tenagophila/S. mansoni interaction. The possibility for applying the knowledge acquired to obtain a biological model for the control of S. mansoni transmission in endemic areas is discussed.

Plasmodium-mosquito interactions, phage display libraries and transgenic mosquitoes impaired for malaria transmission.

Malaria continues to kill millions of people every year and new strategies to combat this disease are urgently needed. Recent advances in the study of the mosquito vector and its interactions with the malaria parasite suggest that it may be possible to genetically manipulate the mosquito in order to reduce its vectorial capacity. Here we review the advances made to date in four areas: (1) the introduction of foreign genes into the mosquito germ line; (2) the characterization of tissue-specific promoters; (3) the identification of gene products that block development of the parasite in the mosquito; and (4) the generation of transgenic mosquitoes impaired for malaria transmission. While initial results show great promise, the problem of how to spread the blocking genes through wild mosquito populations remains to be solved.

Robust gut-specific gene expression in transgenic Aedes aegypti mosquitoes.

Genetic modification of the vectorial capacity of mosquito vectors of human disease requires promoters capable of driving gene expression with appropriate tissue and stage specificity. We report on the characterization in transgenic Aedes aegypti of two mosquito gut-specific promoters. A 1.4-kb DNA fragment adjacent to the 5' end of the coding region of the Ae. aegypti carboxypeptidase (AeCP) gene and a corresponding 3.4-kb DNA fragment at the 5' end of the Anopheles gambiae carboxypeptidase (AgCP) gene were linked to a firefly luciferase reporter gene and introduced into the Ae. aegypti germ line by using Hermes and mariner (Mos1) transposons. Six independent transgenic lines were obtained with the AeCP construct and one with the AgCP construct. Luciferase mRNA and protein were abundantly expressed in the guts of transgenic mosquitoes in four of the six AeCP lines and in the AgCP line. Expression of the reporter gene was gut-specific and reached peak levels at about 24 h post-blood ingestion. The AeCP and AgCP promoters can be used to drive the expression of genes that hinder parasite development in the mosquito gut.

[Hypochondriasis by proxy in children: report of two cases] / Hipocondria por procuração em crianças: relato de dois casos.

OBJECTIVE: To emphasize important aspects for the diagnosis and approach of patients with history of child abuse, related with the Munchausen Syndrome by proxy, and its variant, Hypochondriases by proxy. METHODS: The authors describe two cases followed in the Pediatric Service in a terciary hospital of Salvador, Bahia, in which they found aspects of the medical history and physical examination of these children and in the mothers' behavior compatible with Munchausen by proxy (Factitious disorder by proxy). RESULTS: In the two cases presented, we observed a standard in the clinical history, described as recurrent inpatient and outpatient medical support, through the last three years, without the identification of abnormal results. These facts do not decreased the certainty of the mothers about their children's disease (hypochondriases by proxy). Furthermore we observed cooperation on the part of the children by incorporating the role of the sick one, which made their school and social adaptation more difficult. CONCLUSIONS: The pediatrician must be aware of the diagnosis of Munchausen Syndrome by Proxy and its variation, hypochondriases by proxy, in those cases of unsuitable and recurrent disease, lack of symptoms and signs in the absence of parents, and diagnostic methods with meaningless results. When absolutely certain about diagnosis, the physician must establish the risk for the child (increased mortality in these cases) and suggest psychotherapy for the mother or perpetrator.

Ceftriaxone: una terapéutica antimicrobiana asociada a pseudolitiasis biliae en pediatría / Ceftriaxone: an antimicrobial therapy associated to biliary pseudolithiasis in children

Ceftriaxone es una cefalosporina B-lactamasa resistente,de tercera generación y mucho uso en pediatría pr su amplio espectro de acción. El término pseudolitiasis biliar se debe a la naturaleza reversible de la anormalidad de la vesícula biliar cuando se suspende el ceftriaxone. Presentamos 3 pacientes pediátricos que recibieron ceftriaxone durante el posoperatorio, internados por cuadros clínicos de abdomen agudo y que desarrollaron pseudolitiasis biliar medicamentosa. La cefalosporinafue administrada por vía parental a 100mg/Kg/día, fraccionada en dos dosis a partir del primer día del posoperatorio en promedio durante 11,3 días, sólo un paciente tuvo un cambio del antibiótico por la sospecha precoz de una correlación entre la pseudolitiasis y el ceftriaxone. No hubo diferencia entre sexo, edad o duración de la terapéutica ni cuadro clínico sugestivo de patología biliar. Ningún paciente necesitó ser sometido a colecistectomía y desaparecieron los hallazgos ultrasonográficos al suspender el antibiótico. Las características de esta litiasis"iatrogénica" la transforma en una intidad clínica que necesita ser diagnosticada y evaluada precozmente para evitar intervenciones qurúrgicas innecesarias

Ceftriaxone: una terapéutica antimicrobiana asociada a pseudolitiasis biliae en pediatría / Ceftriaxone: an antimicrobial therapy associated to biliary pseudolithiasis in children

Ceftriaxone es una cefalosporina B-lactamasa resistente,de tercera generación y mucho uso en pediatría pr su amplio espectro de acción. El término pseudolitiasis biliar se debe a la naturaleza reversible de la anormalidad de la vesícula biliar cuando se suspende el ceftriaxone. Presentamos 3 pacientes pediátricos que recibieron ceftriaxone durante el posoperatorio, internados por cuadros clínicos de abdomen agudo y que desarrollaron pseudolitiasis biliar medicamentosa. La cefalosporinafue administrada por vía parental a 100mg/Kg/día, fraccionada en dos dosis a partir del primer día del posoperatorio en promedio durante 11,3 días, sólo un paciente tuvo un cambio del antibiótico por la sospecha precoz de una correlación entre la pseudolitiasis y el ceftriaxone. No hubo diferencia entre sexo, edad o duración de la terapéutica ni cuadro clínico sugestivo de patología biliar. Ningún paciente necesitó ser sometido a colecistectomía y desaparecieron los hallazgos ultrasonográficos al suspender el antibiótico. Las características de esta litiasis"iatrogénica" la transforma en una intidad clínica que necesita ser diagnosticada y evaluada precozmente para evitar intervenciones qurúrgicas innecesarias

Effect of Eucalyptus feeding in the development, survival and reproduction of Tynacantha marginata (Heteroptera: Pentatomidae).

The effect of feeding on Eucalyptus leaves of Tynacantha marginata (Heteroptera: Pentatomidae) was studied in Brazil. The use of this plant as a complementary food source for the predatory bug increased its oviposition period, generation time, and net reproductive rate. Eucalyptus leaves apparently had no influence on the survival and development of T. marginata.

[The knowledge of pediatricians in Salvador, Brazil about measles vaccine]. / Conocimientos de los pediatras de Salvador, Brasil, sobre la vacuna antisarampionosa.

The success of international initiatives for the eradication of measles depends in large part on knowledge about vaccination. In 1992, a cross-sectional study was carried out to evaluate knowledge about measles vaccine among pediatricians in the Brazilian city of Salvador, Bahia. Of the 506 pediatricians residing in the city, 299 (59%) could be located and responded to a questionnaire containing 15 questions that posed hypothetical situations regarding indications and contraindications for this vaccine. The average number of correct responses was 9.3 out of 15, which illustrates the pediatricians' poor knowledge of measles vaccine. Conditions commonly seen in pediatric practice in Brazil--malnutrition, upper respiratory tract infection, diarrhea, and prematurity--were often erroneously considered contraindications for immunization. More than half (62%) of the pediatricians did not know the correct way to administer the vaccine. Knowledge about the vaccine did not differ according to the length of time that had elapsed since the physicians' university training in pediatrics or as a result of working in health centers operated by the State Health Secretariat. University professors, pediatricians with a postgraduate degree, and those pursuing postgraduate studies scored slightly higher than the rest. These results point to the need to strengthen training on measles vaccination in medical schools and programs of continuing education for pediatricians.