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Medically important arboviruses such as dengue, Zika, and chikungunya viruses are primarily transmitted by the globally distributed mosquito Aedes aegypti . Increasing evidence suggests that transmission can be influenced by mosquito viromes. Herein RNA-Seq was used to characterize RNA metaviromes of wild-caught Ae . aegypti from Bangkok (Thailand) and from Cairns (Australia). The two mosquito populations showed a high degree of similarity in their viromes. BLAST searches of assembled contigs suggest up to 27 insect-specific viruses may infect Ae . aegypti , with up to 23 of these currently uncharacterized and up to 16 infecting mosquitoes from both Cairns and Bangkok. Three characterized viruses dominated, Phasi Charoen-like virus, Humaita-Tubiacanga virus and Cell fusing agent virus, and comparisons with other available RNA-Seq datasets suggested infection levels with these viruses may vary in laboratory-reared mosquitoes. As expected, mosquitoes from Bangkok showed higher mitochondrial diversity and carried alleles associated with knock-down resistance to pyrethroids. Blood meal reads primarily mapped to human genes, with a small number also showing homology with rat/mouse and dog genes. These results highlight the wide spectrum of data that can be obtained from such RNA-Seq analyses, and suggests differing viromes may need to be considered in arbovirus vector competence studies.

Background Chikungunya virus (CHIKV) is a mosquito-transmitted, arthritogenic alphavirus that causes sporadic outbreaks of often debilitating rheumatic disease. The recently approved CHIKV vaccine, IXCHIQ, is based on a live-attenuated CHIKV strain (VLA1553), with viraemic vaccine recipients theoretically able to transmit VLA1553 to mosquitoes with ensuing onward transmission. We thus evaluated VLA1553 transmission from artificial blood meals to Aedes albopictus mosquitoes, and onward transmission to mice. Methods Female A. albopictus mosquitoes were fed on defibrinated sheep blood containing wild-type CHIKV (viral titre: 7.50 log 10 CCID 50 /mL) or VLA1553 (viral titres: 7.85, 5.72, 4.58, and 3.79 log 10 CCID 50 /mL). Viral titres in mosquito bodies and saliva were determined using CCID 50 assays 7–8 days after the blood meal. After providing CHIKV or VLA1553 (viral titres ~ 7–8 log 10 CCID 50 /mL) in blood meals to mosquitoes, infected mosquitoes were fed on highly susceptible Irf3/7 −/− mice ( n  = 3 per group). Data were re-analysed using the same reverse transcription quantitative polymerase chain reaction (RT-qPCR) as for an earlier VLA1553 phase 1 clinical trial, to allow correlations between blood meal titres and viraemia in vaccine recipients. Results Mosquito body viral titres were significantly higher ( P  < 0.0001) for CHIKV versus VLA1553-fed mosquitoes at blood meal viral titres of ~ 7–8 log 10 CCID 50 /mL. Mosquito body VLA1553 titres decreased with reducing blood meal titres, but there was no dose-dependent effect on saliva viral titres. No dissemination to salivary glands was seen at blood meal titres ≤ 3.875 log 10 CCID 50 /mL. CHIKV-fed mosquitoes were able to transmit virus, and induce viraemia in, 3/3 Irf3/7 −/− mice via mosquito bites. In contrast, 0/3 Irf3/7 −/− mice became infected after bites from VLA1553-fed mosquitoes. RT-qPCR comparisons with phase 1 clinical data for VLA1553-vaccinated individuals indicated that VLA1553 viraemia was at or below the aforementioned threshold for transmission. Conclusions The evidence presented herein argue that the low viraemia in VLA1553-vaccinated individuals would mitigate against transmission. In addition, replication of VLA1553 in mosquito bodies was also significantly attenuated. Overall, mosquito-borne transmission of VLA1553 from vaccinated individuals to others appears improbable. Graphical Abstract

Mosquito control with bednets, residual sprays or fumigation remains the most effective tool for preventing vector-borne diseases such as malaria, dengue and Zika, though there are no widely used entomological methods for directly assessing its efficacy. Mosquito age is the most informative metric for evaluating interventions that kill adult mosquitoes but there is no simple or reliable way of measuring it in the field. Near-Infrared Spectroscopy (NIRS) has been shown to be a promising, high-throughput method that can estimate the age of mosquitoes. Currently the ability of NIRS to measure mosquito age is biased, and has relatively high individual mosquito measurement error, though its capacity to rigorously monitor mosquito populations in the field has never been assessed. In this study, we use machine learning methods from the chemometric literature to generate more accurate, unbiased estimates of individual mosquito age. These unbiased estimates produce precise population-level measurements, which are relatively insensitive to further increases in NIRS accuracy when feasible numbers of mosquitoes are sampled. The utility of NIRS to directly measure the impact of pyrethroid resistance on mosquito control is illustrated, showing how the technology has potential as a highly valuable tool for directly assessing the efficacy of mosquito control interventions.

Background Chikungunya virus (CHIKV) is a mosquito-borne alphavirus associated with epidemics of acute and chronic arthritic disease in humans. Aedes albopictus has emerged as an important new natural vector for CHIKV transmission; however, mouse models for studying transmission have not been developed. Methods Aedes albopictus mosquitoes were infected with CHIKV via membrane feeding and by using infected adult wild-type C57BL/6 mice. Paraffin sections of infected mosquitoes were analysed by immunofluorescent antibody staining using an anti-CHIKV antibody. CHIKV-infected mosquitoes were used to infect adult C57BL/6 and interferon response factor 3 and 7 deficient (IRF3/7 -/- ) mice. Results Feeding mosquitoes on blood meals with CHIKV titres > 5 log 10 CCID 50 /ml, either by membrane feeding or feeding on infected mice, resulted in  ≥ 50 % of mosquitoes becoming infected. However, CHIKV titres in blood meals  ≥ 7 log 10 CCID 50 /ml were required before salivary glands showed significant levels of immunofluorescent staining with an anti-CHIKV antibody. Mosquitoes fed on blood meals of 7.5 (but not 5.9) log 10 CCID 50 /ml were able efficiently to transmit virus to adult C57BL/6 and IRF3/7 -/- mice, with the latter mice showing overt signs of arthritis post-infection. Conclusions The results provide a simple in vivo model for studying transmission of CHIKV from mosquitoes to mammals and also argue against a resistance barrier to CHIKV infection in adult mice.

I met Margaret Wente's column in the Jan. 6 edition of the Globe and Mail with anger initially, but disappointment has slowly replaced that. Don't all of us work with imperfect and incomplete information at all times? Wente chose to brush her truth with long and broad strokes, with little regard for what the canvas really contains. Danny Williams' fight for these oil revenues gives the province at least a chance for a viable future. Newfoundland and the oil companies that work there pump more than 100 million barrels of oil, worth $3 billion, into the economy on an annual basis. After clawbacks, Newfoundland earns a paltry sum of $30 million to $50 million on an annual basis. Similarly, Alberta earns approximately $6 billion every year. In addition to oil, Newfoundland supplies massive reserves of hydroelectricity to Quebec, which yields an annual estimated profit of $1 billion to their coffers. Of this profit, Newfoundland receives a paltry three per cent. And this situation exists only because Ottawa would not grant Newfoundland the right to export energy across provincial lines.

Efficient as the Japanese language is, gaijin means non- Japanese, lumping the rest of the world together into one pot -- a clearly defined way of looking at the world, I think, an us-versus- them mentality which defines a lot of Japanese culture. No hyphenated labeling of cultural heritage in Japan, no Chinese- Japanese or American-Japanese. Even people with mixed heritage, say a person with a Korean great-grandmother, gets put into the gaijin category.

I decide enough is enough and give in to temptation. I enter McDonald's with trepidation, fears of becoming the \"Ugly Foreigner,\" the kind who storm into quiet tofu restaurants and demand slabs of steak. McDonald's restaurants look the same here as they do back home, even down to the uniforms. I place my order, playing it safe with the standard burger and fries. I ask for a large drink, and get something close to a North American medium. The cashier bows deeply after passing me the tray.

O'Hare, in Chicago, was easier, even with the presence of the camouflaged National Guard, who seemed just to be handing out directions to confused travellers. In Narita, I watched the world go by: people catching flights to Cairo via Malaysia, Sydney via Seoul. A family rushed to catch a plane at the very last minute, arms loaded with souvenirs from Tokyo Disneyland. My first preconception about Japan is shattered: Food prices really are not too high. They seem on par with Canada, once you factor in all the taxes we pay. Some things, though, are outrageously steep. Melons, for example, cost about $40 Cdn. Apples sell for $1 to $1.20 each, but they're big -- a meal in themselves. Shops and restaurants have a wider selection than in Canada and it pays to shop around -- a cup of coffee and a slice of toast, for example, can cost $7 in one coffee shop, but half that across the street.

If the protesters who marched in downtown Kingston last month...

Kevin Chong makes an excellent point about the state of book reviews in Canada (Reviewing the Reviewers, March 31). I do agree with him on some major points -- far too often CanLit books are maple leaf-stamped by reviewers eager to promote some murky sense of nationalism. The question I want to pose that Chong didn't ask is: Who are these reviewers anyway? Or, more bluntly, who do they think they are? A writer works hard, pores over pages and lines, rewrites, drinks too much, re-edits, loses his or her eyesight, rewrites some more.