BackgroundAedes aegypti mosquitoes are the main vectors of dengue viruses to humans. Understanding their biology and interactions with the pathogen are prerequisites for development of dengue transmission control strategies. Mosquito salivary glands are organs involved directly in pathogen transmission to vertebrate hosts. Information on the spatial distribution of gene expression in these organs is expected to assist in the development of novel disease control strategies, including those that entail the release of transgenic mosquitoes with impaired vector competence.ResultsWe report here the hybridization in situ patterns of 30 transcripts expressed in the salivary glands of adult Ae. aegypti females. Distinct spatial accumulation patterns were identified. The products of twelve genes are localized exclusively in the proximal-lateral lobes. Among these, three accumulate preferentially in the most anterior portion of the proximal-lateral lobe. This pattern revealed a salivary gland cell type previously undescribed in Ae. aegypti, which was validated by transmission electron microscopy. Five distinct gene products accumulate in the distal-lateral lobes and another five localize in the medial lobe. Seven transcripts are found in the distal-lateral and medial lobes. The transcriptional product of one gene accumulates in proximal- and distal-lateral lobes. Seven genes analyzed by quantitative PCR are expressed constitutively. The most abundant salivary gland transcripts are those localized within the proximal-lateral lobes, while previous work has shown that the distal-lateral lobes are the most active in protein synthesis. This incongruity suggests a role for translational regulation in mosquito saliva production.ConclusionsTransgenic mosquitoes with reduced vector competence have been proposed as tools for the control of dengue virus transmission. Expression of anti-dengue effector molecules in the distal-lateral lobes of Ae. aegypti salivary glands has been shown to reduce prevalence and mean intensities of viral infection. We anticipate greater efficiency of viral suppression if effector genes are expressed in all lobes of the salivary glands. Based on our data, a minimum of two promoters is necessary to drive the expression of one or more anti-dengue genes in all cells of the female salivary glands.

A cross sectional survey of knowledge, attitude and practices related to house flies among dairy farmers in Punjab, Pakistan AbstractBackground: House flies are of major public health concerns in areas with poor sanitation and hygienic conditions. Unfortunately, sanitation and hygiene have always been ignored in dairy farms particularly in the developing or low-income countries. Management of these flies mostly depends on the awareness regarding associated hazards and protective measures taken by the people to minimize risks associated with flies. The present study therefore explores the knowledge, attitude and practices taken by dairy farmers in Punjab, Pakistan against house flies.Methods: The present study was based on a cross sectional self administered survey to a convenience sample of 173 small scale dairy farmers in four localities -Multan, Lahore, Shorkot and Faisalabad -of Pakistan. The relationships between socio-demographics, knowledge and preventive practices were investigated through logistic regression analysis and chi-square test of association.Results: Considerable number of dairy farmers 71/173 (41.04%) had no idea about the problems associated with house flies. Although 77/173 (44.51%) dairy farmers reported house flies as disease transmitters, only 23 (29.87%) farmers were familiar with diseases and 22 (28.57%) had somewhat idea of the mode of disease transmission. We found a positive association between dairy farmer's education level and overall knowledge of house flies in multivariate analysis. Farmer's education level and knowledge of the house flies breeding sites had a positive association with the adoption of house fly prevention practices by the respondents. However, knowledge of the problems associated with house flies and preventive measures had no association with house fly prevention practices. Conclusion:The present ethnoentomological survey provides information about knowledge, attitude and practices of dairy farmers related to house flies in Punjab, Pakistan. We conclude that the farmers' education level and knowledge of the breeding sites had a positive association with the adoption of prevention practices against house flies. The study also highlights the need of targeting the lack of knowledge of dairy farmers for the successful management of house flies.

The house fly, Musca domestica L., is an important ectoparasite with the ability to develop resistance to insecticides used for their control. Thiamethoxam, a neonicotinoid, is a relatively new insecticide and effectively used against house flies with a few reports of resistance around the globe. To understand the status of resistance to thiamethoxam, eight adult house fly strains were evaluated under laboratory conditions. In addition, to assess the risks of resistance development, cross-resistance potential and possible biochemical mechanisms, a field strain of house flies was selected with thiamethoxam in the laboratory. The results revealed that the field strains showed varying level of resistance to thiamethoxam with resistance ratios (RR) at LC50 ranged from 7.66-20.13 folds. Continuous selection of the field strain (Thia-SEL) for five generations increased the RR from initial 7.66 fold to 33.59 fold. However, resistance declined significantly when the Thia-SEL strain reared for the next five generations without exposure to thiamethoxam. Compared to the laboratory susceptible reference strain (Lab-susceptible), the Thia-SEL strain showed cross-resistance to imidacloprid. Synergism tests revealed that S,S,S-tributylphosphorotrithioate (DEF) and piperonyl butoxide (PBO) produced synergism of thiamethoxam effects in the Thia-SEL strain (2.94 and 5.00 fold, respectively). In addition, biochemical analyses revealed that the activities of carboxylesterase (CarE) and mixed function oxidase (MFO) in the Thia-SEL strain were significantly higher than the Lab-susceptible strain. It seems that metabolic detoxification by CarE and MFO was a major mechanism for thiamethoxam resistance in the Thia-SEL strain of house flies. The results could be helpful in the future to develop an improved control strategy against house flies.

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